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      "titulo" => "Chest Wall CT-Fistulography for Surgical Planning in a Patient With Chronic Granulomatous Disease"
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      <span class="elsevierStyleSections"><span id="sec0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0045">Introduction</span><p id="par0005" class="elsevierStylePara elsevierViewall">Lung cancer&#44; which caused 1&#46;8 million deaths in 2022&#44;<a class="elsevierStyleCrossRef" href="#bib0160"><span class="elsevierStyleSup">1</span></a> is mainly represented by non-small cell lung cancer &#40;NSCLC&#41;&#44; which makes up 85&#37; of cases and has a 5-year survival rate of 20&#37; that varies according to tumor stages&#46;<a class="elsevierStyleCrossRef" href="#bib0165"><span class="elsevierStyleSup">2</span></a> Factors that increase the risk of NSCLC include smoking&#44; environmental exposures&#44; and genetic expression dysregulation due to mutations in genes like <span class="elsevierStyleItalic">EGFR</span>&#44; <span class="elsevierStyleItalic">KRAS</span>&#44; and <span class="elsevierStyleItalic">TP53</span>&#44; among others&#46;<a class="elsevierStyleCrossRef" href="#bib0170"><span class="elsevierStyleSup">3</span></a> NSCLC is mainly histologically divided into adenocarcinoma and squamous cell carcinoma&#46; According to the last European Society for Medical Oncology &#40;ESMO&#41; guidelines surgery remains the mainstay treatment for stages I&#8211;II and resectable stage III NSCLC&#46; After surgery&#44; adjuvant chemotherapy should be offered to patients with resected stages IIB and III NSCLC and can be considered in patients with stage IIA&#46;<a class="elsevierStyleCrossRef" href="#bib0175"><span class="elsevierStyleSup">4</span></a> Management of locally advanced stage &#40;IIIA&#47;B&#41; NSCLC consists of multimodal treatment including surgery&#44; chemotherapy&#44; chemoradiotherapy or immune checkpoint inhibitors&#47;tyrosine kinase inhibitors&#44; depending on disease and patient status and the molecular tumor profile&#46;<a class="elsevierStyleCrossRefs" href="#bib0180"><span class="elsevierStyleSup">5&#44;6</span></a> Despite the treatments used&#44; 30&#8211;35&#37; of patients with NSCLC relapse within five years&#44; mainly due to minimal residual disease &#40;MRD&#41; from resection and high genetic heterogeneity&#46;<a class="elsevierStyleCrossRef" href="#bib0170"><span class="elsevierStyleSup">3</span></a> Tumors harbor driver mutations &#40;i&#46;e&#46; <span class="elsevierStyleItalic">EGFR</span>&#44; <span class="elsevierStyleItalic">TP53</span>&#41; and branch mutations that cause intratumorally heterogeneity&#44; being responsible for tumor resistance and survival&#46;<a class="elsevierStyleCrossRefs" href="#bib0170"><span class="elsevierStyleSup">3&#44;7</span></a> Precision medicine uses next-generation sequencing &#40;NGS&#41; to identify heterogeneity&#44; select treatments&#44; and unveil the origin and course of disease&#46;<a class="elsevierStyleCrossRefs" href="#bib0195"><span class="elsevierStyleSup">8&#44;9</span></a> Nevertheless&#44; among various strategies in NGS&#44; gene panels distinguish themselves for their cost-effectiveness and precision&#46;<a class="elsevierStyleCrossRefs" href="#bib0205"><span class="elsevierStyleSup">10&#44;11</span></a></p><p id="par0010" class="elsevierStylePara elsevierViewall">Liquid biopsy &#40;LB&#41; monitors the real-time evolution of cancer and circulating tumor cells &#40;CTCs&#41; are one of the elements that correlate with MRD and the risk of relapse in lung cancer patients undergoing surgery&#46;<a class="elsevierStyleCrossRefs" href="#bib0215"><span class="elsevierStyleSup">12&#44;13</span></a> However&#44; the phenotypic diversity of CTCs in lung cancer requires a clinically meaningful profile&#46;<a class="elsevierStyleCrossRefs" href="#bib0220"><span class="elsevierStyleSup">13&#44;14</span></a> CTCs reflect the complexity of tissues&#44; and it is possible to identify clones of resistant tumors that cause relapse&#46; The epithelial&#8211;mesenchymal transition marks the presence of diverse phenotypes and metastatic potential regulated by EZH2&#44; associated with NSCLC progression&#44; metastasis&#44; and maintenance of cancer cell characteristics&#46;<a class="elsevierStyleCrossRefs" href="#bib0230"><span class="elsevierStyleSup">15&#44;16</span></a> Screening for CTC&#8722; EZH2&#43; could help stratify the risk of relapse&#47;death and guide treatments&#46;</p><p id="par0015" class="elsevierStylePara elsevierViewall">The primary objective of this study is to investigate the relationship between the molecular profiles of resectable NSCLC tissue and the presence of CTCs that express EZH2 subpopulations&#46; We aim to determine if these molecular profiles and CTC phenotypes can predict clinical outcomes and the likelihood of relapse&#46; Specifically&#44; we hypothesize that certain molecular characteristics are associated with an increased probability of detecting CTCs&#44; which may serve as minimal residual disease &#40;MRD&#41; indicators&#46; Our results will explore these associations to provide clearer insights into how molecular profiles and CTC presence can inform patient prognosis and treatment strategies&#46;</p></span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0050">Material and Methods</span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0055">Study Design and Patients</span><p id="par0020" class="elsevierStylePara elsevierViewall">Conducted between July 2017 and December 2019&#44; this prospective observational cohort study focused on NSCLC patients undergoing surgical treatment &#40;I&#8211;III&#41; at a single tertiary hospital&#46; Surgery consisted of anatomical pulmonary resection and&#44; systematic lymph node dissection for clinical stages II and IIIA and lobe-specific lymph node dissection for stage I&#46; For the LB analyzes&#44; the peripheral blood samples were collected at baseline &#40;CTC1&#41; and one month after surgery &#40;CTC2&#41;&#46; Tumor tissue was extracted during surgery&#44; tumoral and peritumoral samples were obtained through a sterile field and placed in RNA Later tubes &#40;AM7020&#44; ThermoFisher Scientific&#44; Waltham&#44; Massachusetts&#44; USA&#41; for preservation&#46;</p><p id="par0025" class="elsevierStylePara elsevierViewall">The pathologic stage was determined by the 8th edition of the international tumor-node-metastasis &#40;TNM&#41; system and&#44; according to the World Health Organization classification&#44; the histologic subtypes of lung cancer were assigned&#46; After surgery&#44; patients were discussed in the hospital&#39;s thoracic multidisciplinary oncology committee&#46; Adjuvant chemotherapy was indicated in stages IIb and III following the ESMO guidelines&#44; and it was considered in patients with stage IIA&#46;</p><p id="par0030" class="elsevierStylePara elsevierViewall">The follow-up protocol consisted of surveillance every 6 months for the first 3 years&#44; which included a chest and abdominal CT scan&#44; with optional FDG-PET if required&#46; Then&#44; the follow-up was yearly&#46; Clinical outcomes regarding overall survival &#40;OS&#41; and progression-free survival &#40;PFS&#41; were estimated using the Kaplan&#8211;Meier method and both were calculated at 5 years after surgical treatment&#46;</p><p id="par0035" class="elsevierStylePara elsevierViewall">The study&#44; adhering to the Declaration of Helsinki and approved by regional institutional review boards&#44; obtained written informed consent from all participants before enrollment&#46;</p></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0060">gDNA extraction from tissue samples and quantification</span><p id="par0040" class="elsevierStylePara elsevierViewall">Matched tumoral&#47;peritumoral tissue biopsies were collected from patients that underwent surgical treatment&#46; gDNA was extracted using the NucleoSpin&#174; Tissue kit &#40;740952&#46;50&#44; Macherey-Nagel&#44; D&#252;ren&#44; Germany&#41;&#44; following the manufacturer&#39;s instructions&#46; Briefly&#44; 25<span class="elsevierStyleHsp" style=""></span>mg of tissue was used for the extraction&#46; Tissues were homogenized using the TissueLyser LT instrument &#40;Qiagen&#44; Venlo&#44; Netherlands&#41; for 5<span class="elsevierStyleHsp" style=""></span>min at 50<span class="elsevierStyleHsp" style=""></span>oscillations&#47;s&#46; After that&#44; samples were lysed with Proteinase K solution at a final concentration of 2&#46;7<span class="elsevierStyleHsp" style=""></span>mg&#47;ml at 56<span class="elsevierStyleHsp" style=""></span>&#176;C overnight in a shaker&#46; After purification and precipitation&#44; DNA was bound to a silica column&#44; washed twice&#44; and the column was dried out&#46; DNA was eluted in a final volume of 75<span class="elsevierStyleHsp" style=""></span>&#956;l&#46; Concentration was measured using a NanoDrop&#8482; 2000 instrument &#40;ThermoFisher&#41; at a wavelength of 260<span class="elsevierStyleHsp" style=""></span>nm&#46;</p></span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0065">Library Preparation</span><p id="par0045" class="elsevierStylePara elsevierViewall">Sequencing libraries were constructed using the KAPA HyperCap workflow v3&#46;0 &#40;Roche&#44; Basel&#44; Switzerland&#41; with a customized gene panel&#44; following the manufacturer&#39;s protocol&#46; This customized panel was developed to cover 50 genes &#40;exons&#41; involved in four different biological processes&#58; tumor promotion &#40;<span class="elsevierStyleItalic">ROS1</span>&#44; <span class="elsevierStyleItalic">ALK</span>&#44; <span class="elsevierStyleItalic">TP53</span>&#44; <span class="elsevierStyleItalic">APC</span>&#44; <span class="elsevierStyleItalic">RB1</span>&#44; <span class="elsevierStyleItalic">RET</span>&#44; <span class="elsevierStyleItalic">FLT1</span>&#44; <span class="elsevierStyleItalic">PTEN</span>&#44; <span class="elsevierStyleItalic">STK11</span>&#44; <span class="elsevierStyleItalic">TET2</span>&#44; <span class="elsevierStyleItalic">CTNNB1</span>&#44; <span class="elsevierStyleItalic">MLH1</span>&#44; <span class="elsevierStyleItalic">EGFR</span>&#44; <span class="elsevierStyleItalic">DDR2</span>&#44; <span class="elsevierStyleItalic">SLC22A18</span>&#44; <span class="elsevierStyleItalic">AKT1</span>&#41;&#44; Chronic Obstructive Pulmonary Disease &#40;COPD&#41; associated genes &#40;<span class="elsevierStyleItalic">ADAMTSL3</span>&#44; <span class="elsevierStyleItalic">ADAM19</span>&#44; <span class="elsevierStyleItalic">PTCH1</span>&#44; <span class="elsevierStyleItalic">ADGRG6</span>&#44; <span class="elsevierStyleItalic">HTR4</span>&#44; <span class="elsevierStyleItalic">FAM13A</span>&#44; <span class="elsevierStyleItalic">CHRNB4</span>&#44; <span class="elsevierStyleItalic">RIN3</span>&#44; <span class="elsevierStyleItalic">TGFB2</span>&#44; <span class="elsevierStyleItalic">IREB2</span>&#44; <span class="elsevierStyleItalic">SERPINA1</span>&#44; <span class="elsevierStyleItalic">CHRNA3</span>&#44; <span class="elsevierStyleItalic">STN1</span>&#44; <span class="elsevierStyleItalic">CHRNA5</span>&#44; <span class="elsevierStyleItalic">AGER</span>&#41;&#44; lung development &#40;<span class="elsevierStyleItalic">FGF10</span>&#44; <span class="elsevierStyleItalic">FOXA1</span>&#44; <span class="elsevierStyleItalic">FOXA2</span>&#44; <span class="elsevierStyleItalic">FOXP2</span>&#44; <span class="elsevierStyleItalic">FOXP4</span>&#44; <span class="elsevierStyleItalic">HOXA2</span>&#44; <span class="elsevierStyleItalic">HOXA5</span>&#44; <span class="elsevierStyleItalic">HOXB2</span>&#44; <span class="elsevierStyleItalic">PITX2</span>&#44; <span class="elsevierStyleItalic">WNT2</span>&#44; <span class="elsevierStyleItalic">ALX1</span>&#44; <span class="elsevierStyleItalic">NKX2-1</span>&#41; and inflammation &#40;<span class="elsevierStyleItalic">CD163</span>&#44; <span class="elsevierStyleItalic">S100A4</span>&#44; <span class="elsevierStyleItalic">NLRP3</span>&#44; <span class="elsevierStyleItalic">MUC5AC</span>&#44; <span class="elsevierStyleItalic">EGR1</span>&#44; <span class="elsevierStyleItalic">TLR4</span>&#44; <span class="elsevierStyleItalic">FPR2</span>&#41;&#44; spanning approximately 140<span class="elsevierStyleHsp" style=""></span>kb&#46; It was tested on I&#8211;IIIB stage NSCLC to elucidate the contribution of non-tumoral gene mutations on cancer disease development&#44; intercepting cancer&#46; Libraries were quantified with Qubit 4&#46;0 Fluorometer using dsDNA HS Assay kit&#46; Individual libraries were diluted and pooled together in equal amounts &#40;by mass&#44; equimolarity&#41; for obtaining equal numbers of sequencing reads &#40;HiSeq X System&#44; Illumina&#44; San Diego&#44; California&#44; USA&#41;&#46;</p></span><span id="sec0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0070">Sequencing Data Analysis</span><p id="par0050" class="elsevierStylePara elsevierViewall">Once FASTQ files were demultiplexed and retrieved from the sequencer&#44; files were ready for data processing&#46; The GATK best practices-based on the Somatic Variant Pipeline &#40;<a href="https://www.clinbioinfosspa.es/content/somatic-variant-pipeline">https&#58;&#47;&#47;www&#46;clinbioinfosspa&#46;es&#47;content&#47;somatic-variant-pipeline</a>&#41; from the Clinical Bioinformatics Area from Fundaci&#243;n Progreso y Salud &#40;Andalusian Govern &#8211; Health and Family Counseling&#41; were followed&#46; Briefly&#44; raw data quality was evaluated using the FastQC tool &#40;v0&#46;11&#46;9&#44; Babraham Bioinformatics&#44; Cambridge&#44; UK&#41;&#46; High quality reads were aligned against the human reference genome &#40;GRCh38&#41; using Burrow&#8211;Wheeler Aligner algorithm &#40;<a href="https://github.com/lh3/bwa">https&#58;&#47;&#47;github&#46;com&#47;lh3&#47;bwa</a>&#41;&#46; After filtering steps&#44; tumor with matched normal mode from Mutect2 pipeline from GATK &#40;<a href="https://gatk.broadinstitute.org/hc/en-us">https&#58;&#47;&#47;gatk&#46;broadinstitute&#46;org&#47;hc&#47;en-us</a>&#41; was run for variant calling&#46; Generated filtered VCFs were annotated using VEP &#40;v99&#44; <a href="https://www.ensembl.org/info/docs/tools/vep/index.html">https&#58;&#47;&#47;www&#46;ensembl&#46;org&#47;info&#47;docs&#47;tools&#47;vep&#47;index&#46;html</a>&#41;&#46; Clinical significance was assessed by following the American College of Medical Genetics &#40;ACMG&#41; criteria&#46; R software &#40;v4&#46;2&#46;1&#41; and RStudio &#40;2022&#46;12&#46;0<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>353 version&#41; were used for file parsing and for plot generation&#46; Maftools<a class="elsevierStyleCrossRef" href="#bib0240"><span class="elsevierStyleSup">17</span></a> was used for oncoplot generation and somatic interactions plot&#46;</p></span><span id="sec0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0075">Collection of Blood Samples&#46; Enrichment and Isolation of CTCs</span><p id="par0055" class="elsevierStylePara elsevierViewall">Peripheral blood &#40;10<span class="elsevierStyleHsp" style=""></span>ml&#41; was collected in EDTA BD Vacutainer&#174; tubes and stored at room temperature for processing within the next 4<span class="elsevierStyleHsp" style=""></span>h&#46; CTCs were isolated by density gradient centrifugation&#44; following a previously described protocol&#46;<a class="elsevierStyleCrossRef" href="#bib0215"><span class="elsevierStyleSup">12</span></a> Briefly&#44; blood samples were centrifuged by density gradient with Ficoll Histopaque&#174;-1119 &#40;density<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>1&#46;119<span class="elsevierStyleHsp" style=""></span>g&#47;ml&#41; &#40;Merck&#44; Darmstadt&#44; Germany&#41; for 45<span class="elsevierStyleHsp" style=""></span>min at 700<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleItalic">g</span> with no break&#46; The buffy coat where CTCs and PBMCs are located was carefully extracted&#46; Isolation of CTCs was performed by positive immunomagnetic selection using the Carcinoma Cell Enrichment kit &#40;130-108-339&#44; Miltenyi Biotec&#44; Bergisch Gladbach&#44; Germany&#41;&#46; This selection is based on a multi-cytokeratin antibody &#40;CK3-6H5&#41;&#44; that recognizes the cytoplasmic cytokeratins 7 and 8&#46; Cellular samples were spun down onto slides that were stained and characterized by immunofluorescence&#46;</p></span><span id="sec0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0080">CTC Characterization by Immunofluorescence</span><p id="par0060" class="elsevierStylePara elsevierViewall">The working area was marked with a hydrophobic pen &#40;Dako&#41;&#46; Each sample was hydrated with PBS 1&#215; for 5<span class="elsevierStyleHsp" style=""></span>min and incubated with anti-cytokeratin-FITC antibody&#44; against cytokeratins 7 and 8 &#40;130-060-301&#44; Miltenyi Biotec&#41;&#44; diluted 1&#58;100 in PBS 1&#215; &#8211; Tween 0&#46;1&#37;<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>BSA 2&#37; &#40;working solution&#41; for 45<span class="elsevierStyleHsp" style=""></span>min in the dark at room temperature&#46; After this point&#44; darkness was always maintained&#46; After this incubation&#44; samples were washed with PBS 1&#215; for 5<span class="elsevierStyleHsp" style=""></span>min&#46; A blockade was performed with goat serum diluted at 1&#215; in working solution for 1<span class="elsevierStyleHsp" style=""></span>h&#46; To detect the EZH2 protein&#44; primary rabbit anti-EZH2 antibody &#40;&#35;5246S&#44; Cell Signaling Technology&#41; was diluted 1&#58;100 in the working solution and was incubated for 45<span class="elsevierStyleHsp" style=""></span>min&#46; Then&#44; samples are washed in PBS 1&#215; for 5<span class="elsevierStyleHsp" style=""></span>min&#46; Secondary goat anti-rabbit IgG &#40;Alexa Fluor 555&#44; ThermoFisher&#41; was diluted 1&#58;200 in the working solution and was incubated for 45<span class="elsevierStyleHsp" style=""></span>min to reveal EZH2 expression&#46; Nuclei were counterstained using Hoechst33342 &#40;Fluxion&#41; at 1&#215;&#46; Slides were mounted using SlowFade&#8482; Diamond Antifade Mountant &#40;S36972&#44; ThermoFisher&#41; for visualization in Zeiss LSM 710 confocal microscope&#46;</p><p id="par0065" class="elsevierStylePara elsevierViewall">Evaluation of EZH2 in CTCs was analyzed after previous evaluation on A549 control cell line &#40;CCL-185&#8482;&#44; ATCC&#44; Manassas&#44; Virginia&#44; USA&#41; for antibody specificity &#40;<a class="elsevierStyleCrossRef" href="#sec0110">Supplementary Fig&#46; 1</a>&#41;&#46; Because of the heterogeneity observed in patients with CTCs&#44; the percentage expression of EZH2 was calculated as the number of EZH2&#43; CTCs divided by the total number of CTCs&#46; This calculation aimed to quantify the heterogeneity of CTCs within patients&#46;</p></span><span id="sec0045" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0085">Statistical Analysis</span><p id="par0070" class="elsevierStylePara elsevierViewall">CTCs were assessed both as a continuous variable &#40;number of CTCs per 10<span class="elsevierStyleHsp" style=""></span>ml of blood&#41; and as a binary variable &#40;presence&#47;absence and a cut-off of &#8805;2 CTCs per 10<span class="elsevierStyleHsp" style=""></span>ml&#41;&#46; Cut-offs to distinguish low&#47;high CTC numbers and EZH2 expression were determined using the Cut-off Finder web application &#40;fit for mixture model&#41;&#46;<a class="elsevierStyleCrossRef" href="#bib0245"><span class="elsevierStyleSup">18</span></a> Bivariate analysis examined the relation between clinical features and CTC presence&#47;absence&#46; Clinical variables were evaluated for normality using the Shapiro&#8211;Wilk test&#46;</p><p id="par0075" class="elsevierStylePara elsevierViewall">The clinical characteristics were evaluated by Chi-squared &#40;Gender&#44; Procedure&#44; Histology&#44; Respiratory history&#44; Stage&#44; Adjuvancy&#44; Recurrence and Vital status&#41;&#44; Fisher&#39;s exact test &#40;Tobacco&#44; Resection&#44; Lymph node status&#44; Type of adjuvancy&#44; Site of recurrence&#44; and Cause of death&#41;&#44; <span class="elsevierStyleItalic">t</span>-test &#40;Age&#41; and Mann&#8211;Whitney <span class="elsevierStyleItalic">U</span> test &#40;PET &#8211; SUVmax&#8211;&#44; Tumor size &#40;cm&#41;&#44; overall survival &#8211; OS and progression-free survival &#8211; PFS&#44; both in months&#41;&#46; Parametric variables are expressed as mean with standard deviation and non-parametric variables as median with interquartile range or down&#47;upper quartile &#40;<span class="elsevierStyleItalic">Q</span>1&#44; <span class="elsevierStyleItalic">Q</span>3&#41;&#46;</p><p id="par0080" class="elsevierStylePara elsevierViewall">Wilcoxon Signed-Rank test compared the median of CTC numbers and EZH2 expression between sampling points&#44; to study the dynamics of these variables&#46; Univariate Kaplan&#8211;Meier &#40;log-rank test&#41; analyzed the influence of CTC and EZH2 cut-offs on PFS and OS&#46; Fisher&#39;s exact test evaluated somatic interactions between gene pairs&#44; while <span class="elsevierStyleItalic">t</span>-tests and Fisher&#39;s exact test were further used to compare mutation profiles concerning recurrence and CTC presence&#44; respectively&#46;</p><p id="par0085" class="elsevierStylePara elsevierViewall">Statistical analyses were performed using IBM SPSS Statistics &#40;version 25&#46;0 for MacOS&#44; IBM Corp&#46;&#44; Armonk&#44; NY&#44; USA&#41;&#44; GraphPad Prism &#40;version 9&#46;0&#46;2 for MacOS&#44; GraphPad software&#44; La Jolla&#44; CA&#44; USA&#41;&#44; and R software &#40;v4&#46;2&#46;1&#41; and RStudio &#40;2023&#46;03&#46;0<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>386 version&#41;&#44; with a <span class="elsevierStyleItalic">p</span>-value<span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05 determining statistical significance&#46;</p></span></span><span id="sec0050" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0090">Results</span><p id="par0090" class="elsevierStylePara elsevierViewall">A total of 76 resectable NSCLC patients were enrolled in the study&#46; A descriptive analysis of the cohort is shown in <a class="elsevierStyleCrossRef" href="#sec0110">Supplementary Table 1</a>&#46; Most patients were males &#40;60&#46;5&#37;&#41; and smokers &#40;81&#46;6&#37;&#41;&#44; with a mean age of 66 years&#46; Lobectomy was the main pulmonary resection performed &#40;77&#46;6&#37;&#41; by Video Assisted Thoracic Surgery &#40;VATS&#41; approach &#40;58&#46;0&#37;&#41;&#46; During the follow-up&#44; 20 patients presented cancer recurrence &#40;26&#46;3&#37;&#41;&#46; The median time of PFS was 21 months&#46; The histological evaluation of NSCLC was conducted using hematoxylin&#8211;eosin staining&#46; The specific histologic types &#40;adenocarcinoma or squamous&#41; were assessed by a pathologist &#40;<a class="elsevierStyleCrossRef" href="#sec0110">Supplementary Fig&#46; 2</a>&#41;&#46;</p><span id="sec0055" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0095">Detection of Somatic Variants in gDNA From Matched Tumoral&#8211;Peritumoral Tissue Biopsies</span><p id="par0095" class="elsevierStylePara elsevierViewall">Out of the total of 76 patients&#44; only 27 were eligible for inclusion in the NGS analyses due to meeting the required quality standards for library preparation&#46; Average concentration of extracted gDNA was 316&#46;53<span class="elsevierStyleHsp" style=""></span>ng&#47;&#956;L &#40;range&#58; 43&#46;85&#8211;946&#46;85<span class="elsevierStyleHsp" style=""></span>ng&#47;&#956;L&#41;&#46; Minimal input ensured for library preparation was 500<span class="elsevierStyleHsp" style=""></span>ng&#46; After library preparation&#44; sequencing yielded a mean coverage of approximately 500&#215; along all the samples&#46; From the 27 patients sequenced&#44; only 4 did not pass variant calling filters and they were removed from subsequent analysis&#46; The total number of variants identified for the 23 remaining patients were 143&#44; with an average of 6 variants per patient &#40;range&#58; 1&#8211;25&#41;&#46; Most of the variants &#40;61&#46;5&#37;&#59; 88&#47;143&#41; were non-coding and SNPs &#40;91&#46;6&#37;&#41;&#44; but only 34&#46;3&#37; &#40;49&#47;143&#41; were known variants&#46; Variants were submitted to ClinVar &#40;Accession numbers&#58; SCV003806285&#8211;SCV003806424&#41;&#46; Regarding the clinical significance of the variants&#44; 9&#46;8&#37; &#40;14&#47;143&#41; were benign&#47;likely benign&#44; 83&#46;2&#37; &#40;119&#47;143&#41; were variants of uncertain significance &#40;VUS&#41; and 7&#46;0&#37; &#40;10&#47;143&#41; were pathogenic&#47;likely pathogenic &#40;<a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>A&#41;&#46; Oncoplot in <a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>B summarizes data for the mutational profiles of each patient and gene&#46; The number of mutations for each patient and the number of samples &#40;patients&#41; affected by mutations in that given gene are represented on both top and right plots&#44; respectively&#46; The top 5 most highly mutated genes were&#58; <span class="elsevierStyleItalic">TP53</span>&#44; <span class="elsevierStyleItalic">FLT1</span>&#44; <span class="elsevierStyleItalic">MUC5AC</span>&#44; <span class="elsevierStyleItalic">EGFR</span> and <span class="elsevierStyleItalic">NLRP3</span>&#44; presented in 43&#37;&#44; 39&#37;&#44; 39&#37;&#44; 26&#37; and 26&#37; of the samples&#44; respectively&#46; Cancer stage&#44; presence&#47;absence of CTCs&#44; expression of EZH2 and mutational signatures &#40;transitions and transversions&#41; are also provided for each patient &#40;<a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>B&#41;&#46;</p><elsevierMultimedia ident="fig0005"></elsevierMultimedia><p id="par0100" class="elsevierStylePara elsevierViewall">Full description of the pathogenic&#44; likely pathogenic variants and hot VUS found is described in <a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#46; Both PCT74 and PCT75 patients recurred and had primary point mutations in <span class="elsevierStyleItalic">EGFR</span> and <span class="elsevierStyleItalic">ROS1</span>&#44; respectively&#44; and both had secondary point mutations in <span class="elsevierStyleItalic">TP53</span> &#40;according to allele frequencies&#41;&#46; Contrarily&#44; patients PCT33 and PCT76&#44; had primary mutations in <span class="elsevierStyleItalic">TP53</span> and secondary mutations in <span class="elsevierStyleItalic">RB1</span>&#44; and they did not have recurrence&#44; suggesting that secondary mutations in <span class="elsevierStyleItalic">TP53</span> might have a stronger impact on disease recurrence&#46;</p><elsevierMultimedia ident="tbl0005"></elsevierMultimedia></span><span id="sec0060" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0100">Analyze Mutually Exclusive and Co-occurring Events</span><p id="par0105" class="elsevierStylePara elsevierViewall">Studying if a pair of genes were mutated simultaneously or if they were mutually exclusive&#44; interactions of the somatic variants were analyzed&#46; The top 5 most significant pairs of genes with co-occurring mutations were&#58; <span class="elsevierStyleItalic">CD163-TLR4</span>&#44; <span class="elsevierStyleItalic">FGF10-FOXP2</span>&#44; <span class="elsevierStyleItalic">ADAMTSL3-FLT1</span>&#44; <span class="elsevierStyleItalic">ADAMTSL3-MUC5AC</span> and <span class="elsevierStyleItalic">MUC5AC-NLRP3</span> with <span class="elsevierStyleItalic">p</span>-values of 0&#46;0056&#44; 0&#46;0118&#44; 0&#46;0142&#44; 0&#46;0144&#44; 0&#46;0183&#44; respectively&#46; There was only one significant pair of genes with mutually exclusive mutations&#58; <span class="elsevierStyleItalic">RIN3-TP53</span> &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;0457&#41;&#46; Interactions among all the genes are represented in <a class="elsevierStyleCrossRef" href="#fig0010">Fig&#46; 2</a>&#46; Detailed data of significant pairs of genes highlighted in <a class="elsevierStyleCrossRef" href="#fig0010">Fig&#46; 2</a> can be found in <a class="elsevierStyleCrossRef" href="#sec0110">Supplementary Table 2</a> with additional information for all the significant pairs of genes&#46;</p><elsevierMultimedia ident="fig0010"></elsevierMultimedia><p id="par0110" class="elsevierStylePara elsevierViewall">Most variants were VUS &#40;83&#46;2&#37;&#41; and as they might be reclassified in the future as either pathogenic&#47;likely pathogenic &#40;P&#41; or benign&#47;likely benign &#40;B&#41;&#44; we studied their impact on recurrence by grouping them with either P or B&#46; Two comparisons were done&#58; P<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>VUS &#40;pathogenic variants and VUS&#41; and VUS<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>B &#40;VUS and benign variants&#41;&#46; P<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>VUS showed significant differences &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;0404&#41; between no-recurrence and recurrence patients&#44; with a mean number of variants of 4&#46;8 &#40;standard error of the mean &#8211; SEM<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;65&#41; in the no-recurrence group versus a mean of 11 &#40;SEM<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>5&#46;54&#41; variants in the recurrence group&#46; On the other hand&#44; VUS<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>B showed no significant differences &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;0597&#41; between no-recurrence &#40;mean of 5&#44; SEM<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;71&#41; and recurrence patients &#40;mean of 11&#44; SEM<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>7&#46;09&#41; &#40;<a class="elsevierStyleCrossRef" href="#fig0015">Fig&#46; 3</a>&#41;&#46;</p><elsevierMultimedia ident="fig0015"></elsevierMultimedia></span><span id="sec0065" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0105">CTCs&#58; Isolation&#44; Characterization&#44; and Prognostic Role</span><p id="par0115" class="elsevierStylePara elsevierViewall">The possible correlation between the clinical&#8211;pathological characteristics of the patients and the presence or absence of CTCs &#40;based on cytokeratin&#44; CK&#44; expression&#41; was investigated before surgery &#40;CTC1&#44; <a class="elsevierStyleCrossRef" href="#tbl0010">Table 2</a>&#41; and one month after surgery &#40;CTC2&#44; <a class="elsevierStyleCrossRef" href="#sec0110">Supplementary Table 3</a>&#41;&#46; The presence of CTC before surgery &#40;CTC1&#41; was significantly correlated with age&#44; lymph node status &#40;N&#41;&#44; tumor stage&#44; and adjuvant treatment&#46; Regarding tumor stage&#44; most patients with stages II and III were CTC&#8722; &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;0300&#41;&#46; Focused on N status&#44; most patients with lymph node metastasis were CTC&#8722; &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;0320&#41;&#46; Consequently&#44; it was observed that most CTC&#8722; patients received adjuvant treatment compared with the CTC&#43; patients &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;0040&#41;&#46;</p><elsevierMultimedia ident="tbl0010"></elsevierMultimedia><p id="par0120" class="elsevierStylePara elsevierViewall">At CTC1&#44; CTCs were detected in 26&#47;76 patients &#40;34&#46;2&#37;&#41; with a median number of 3 in those CTCs&#43; patients &#40;IQR<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>3&#44; range<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>1&#8211;8&#41; meanwhile&#44; at CTC2 the presence of CTCs was detected in only 3&#47;76 patients &#40;3&#46;9&#37;&#41; with a median number of 2 CTCs in those CTCs&#43; patients &#40;IQR<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>3&#44; range<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>1&#8211;4&#41;&#46; In addition&#44; the dynamics of CTCs between the two time points&#44; CTC1 and CTC2&#44; were also analyzed&#46; We observed a significant decrease in the number of CTCs because of the surgery &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;0001&#44; Wilcoxon test&#41; &#40;<a class="elsevierStyleCrossRef" href="#fig0020">Fig&#46; 4</a>A&#41;&#46;</p><elsevierMultimedia ident="fig0020"></elsevierMultimedia><p id="par0125" class="elsevierStylePara elsevierViewall">The expression of EZH2 on CTCs was measured &#40;see CTC characterization by immunofluorescence methods Section&#41; at CTC1 in the 26 CTC&#43; patients&#46; We found that 13&#47;26 patients were EZH2&#43; &#40;50&#46;0&#37;&#41;&#44; with a median percentage of expression of 6&#46;25&#37; &#40;IQR&#58; 54&#46;17&#37;&#44; range&#58; 0&#8211;100&#37;&#41;&#46; At CTC2&#44; EZH2 was detected in 2&#47;3 CTC&#43; patients &#40;66&#46;67&#37;&#41;&#44; with a median percentage of expression of EZH2 of 100&#37; &#40;IQR&#58; 100&#37;&#44; range&#58; 0&#8211;100&#37;&#41;&#46; Expression of EZH2 showed no significant differences between CTC1 and CTC2 &#40;<a class="elsevierStyleCrossRef" href="#fig0020">Fig&#46; 4</a>B&#41;&#46;</p><p id="par0130" class="elsevierStylePara elsevierViewall">The prognostic role of circulating tumor cells &#40;CTCs&#41; for PFS and OS was evaluated using either the presence&#47;absence of CTCs or a cut-off of \n
      &lt;!--&#8805;2 CTCs at CTC1. No significant differences were found for OS or PFS using Kaplan&#8211;Meier analysis for the presence/absence of CTCs (--&gt;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;5430 and <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;9070&#44; respectively&#59; see <a class="elsevierStyleCrossRef" href="#sec0110">Supplementary Fig&#46; 3</a>&#41;&#46; Using the CTC cut-off of \n
      &lt;!--&#8805;2 slightly improved both OS and PFS, but again, no significant differences were observed (--&gt;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;1920 and <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;4380&#44; respectively&#41;&#46; OS was measured at 5 years since surgical treatment&#46;</p><p id="par0135" class="elsevierStylePara elsevierViewall">Interestingly&#44; when evaluating EZH2 expression in the CTCs using a cut-off of &#60;70&#37; or &#8805;70&#37; for the previously calculated percentage of expression &#40;EZH2low&#47;EZH2high&#44; respectively&#41;&#44; significant differences in terms of OS were observed at CTC1 &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;0180&#44; Log-Rank&#44; Mantel&#8211;Cox&#41; with EZH2high patients having an 87&#37; greater risk of death&#46; In terms of PFS no significant differences were found for EZH2 characterization &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;9130&#41; &#40;<a class="elsevierStyleCrossRef" href="#fig0025">Fig&#46; 5</a>C and D&#41;&#46;</p><elsevierMultimedia ident="fig0025"></elsevierMultimedia><p id="par0140" class="elsevierStylePara elsevierViewall">At CTC2 neither presence&#47;absence of CTCs nor CTC2low&#47;CTC2high showed significant differences in terms of PFS or OS &#40;<a class="elsevierStyleCrossRef" href="#sec0110">Supplementary Fig&#46; 4</a>&#41;&#46; At this point&#44; Kaplan&#8211;Meier curves could not be calculated for EZH2low&#47;EZH2high in CTCs&#43; patients due to the very small sample size&#46;</p></span><span id="sec0070" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0110">Integrative Analysis of Tissue Molecular Profiles and Presence of CTCs</span><p id="par0145" class="elsevierStylePara elsevierViewall">Presence or absence of CTCs at baseline was analyzed based on the tissue molecular profiles&#46; While we did not find statistical differences between the presence or absence of CTCs and VUS in the studied genes&#44; we did observe a noteworthy trend&#46; There were two out of three patients without CTCs in the follow-up but with VUS in <span class="elsevierStyleItalic">ADAM19</span> that locally recurred &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;1739&#41;&#46; The only patient with CTCs &#40;both at baseline and the follow-up&#41; and VUS in <span class="elsevierStyleItalic">ADAM19</span> did not recur at time of these analyses &#40;<a class="elsevierStyleCrossRef" href="#sec0110">Supplementary Table 4</a>&#41;&#46;</p></span></span><span id="sec0075" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0115">Discussion</span><p id="par0150" class="elsevierStylePara elsevierViewall">This study examines biological data from different approaches in stage I-IIIB NSCLC patients&#44; evaluating the mutational profile of lung tissue with a customized panel of 50 genes and CTCs&#46; Regarding the prognostic role of CTCs&#44; despite this study including a small population&#44; patients with a high percentage of CTC&#8722; EZH2&#43; showed poorer OS&#46; Our recent results are consistent with previous data associating the expression of the EZH2 protein in breast cancer with metastatic recurrence and worse PFS and OS in this study<a class="elsevierStyleCrossRefs" href="#bib0250"><span class="elsevierStyleSup">19&#8211;21</span></a> and in lung cancer with tumor colonization&#46;<a class="elsevierStyleCrossRef" href="#bib0235"><span class="elsevierStyleSup">16</span></a> Due to the essential and heterogeneous role of EZH2 in regulating various sets of genes in cancer&#44; many clinical trials have emerged to test inhibitors against this target&#46;<a class="elsevierStyleCrossRef" href="#bib0265"><span class="elsevierStyleSup">22</span></a> However&#44; our results also suggest the importance of characterizing the CTCs to identify the aggressiveness of cell subpopulations&#46;<a class="elsevierStyleCrossRefs" href="#bib0270"><span class="elsevierStyleSup">23&#8211;25</span></a> Interestingly&#44; we observed that patients with negative lymph node status &#40;pN0&#41; but positive for circulating tumor cells &#40;CTC&#41; had a shorter OS&#46; These preliminary findings suggest the presence of occult metastatic disease&#46; These results are consistent with previous studies&#44; which have shown that the shedding of tumor cells can occur through pathways independent of lymphatic drainage&#44; indicating that the presence of CTCs in N0 breast cancer predicts disease relapse&#46;<a class="elsevierStyleCrossRef" href="#bib0285"><span class="elsevierStyleSup">26</span></a></p><p id="par0155" class="elsevierStylePara elsevierViewall">In our study&#44; the molecular profiling found an association between <span class="elsevierStyleItalic">RIN3</span> and <span class="elsevierStyleItalic">TP53</span>&#44; which might imply that <span class="elsevierStyleItalic">RIN3</span> variants could induce lung cancer despite the initial normal function of <span class="elsevierStyleItalic">TP53</span>&#46; The role of <span class="elsevierStyleItalic">RIN3</span> regulating tyrosine kinase ABL relates to cytoskeletal remodeling&#44; which affects processes crucial for the spread of lung cancer&#46;<a class="elsevierStyleCrossRef" href="#bib0290"><span class="elsevierStyleSup">27</span></a> A more extensive study integrating COPD and lung cancer patient data may elucidate <span class="elsevierStyleItalic">RIN3&#39;s</span> role in cancer development&#46;</p><p id="par0160" class="elsevierStylePara elsevierViewall">On the other hand&#44; our results suggest that the presence of secondary point mutations affecting <span class="elsevierStyleItalic">TP53</span> together with <span class="elsevierStyleItalic">EGFR</span> mutations is associated with the relapse of the patients&#46; Despite having only two patients with these characteristics&#44; this result aligns with previous works&#44; which have associated concomitant <span class="elsevierStyleItalic">TP53</span> mutations and <span class="elsevierStyleItalic">EGFR</span> mutations with OS and response to treatment&#46; For example&#44; in the study by X&#46; Le et al&#46;&#44; the impact of <span class="elsevierStyleItalic">TP53</span> co-mutations on clinical outcomes in patients with <span class="elsevierStyleItalic">EGFR</span>-mutant advanced NSCLC was examined&#46; This observational study used data from a de-identified database and found that patients with <span class="elsevierStyleItalic">TP53</span> co-mutations had significantly shorter real-world PFS and OS compared to those with wild-type <span class="elsevierStyleItalic">TP53</span> tumors&#46;<a class="elsevierStyleCrossRef" href="#bib0295"><span class="elsevierStyleSup">28</span></a></p><p id="par0165" class="elsevierStylePara elsevierViewall">Furthermore&#44; genetic variants co-occurring in <span class="elsevierStyleItalic">ADAMTSL3</span> and <span class="elsevierStyleItalic">FLT-1</span> &#40;<span class="elsevierStyleItalic">VEGFR-1</span>&#41; suggest involvement in tumor dissemination and prognosis&#44;<a class="elsevierStyleCrossRef" href="#bib0300"><span class="elsevierStyleSup">29</span></a> although the study could not establish their link to CTC presence due to sample size limitations&#46; Investigating <span class="elsevierStyleItalic">ADAMTSL3-FLT1</span> variants in more extensive studies may uncover lung cancer development after COPD&#46;</p><p id="par0170" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">NLRP3</span> and <span class="elsevierStyleItalic">MUC5AC</span>&#44; highly mutated genes in our population&#44; might indicate a potential link to aggressive lung adenocarcinoma and poor outcomes in COPD patients&#44;<a class="elsevierStyleCrossRefs" href="#bib0305"><span class="elsevierStyleSup">30&#44;31</span></a> suggesting their mutational presence could identify COPD patients prone to developing cancer&#46;</p><p id="par0175" class="elsevierStylePara elsevierViewall">Despite the limitations of the small population size&#44; our study observed that baseline tissue variants could predict recurrence better than CTCs&#46; Patients experiencing relapse expressed more potentially impactful &#40;P<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>VUS&#41; variants compared to non-relapsed patients&#46; Less studied genes in the panel contributed to a higher rate of VUS&#44; highlighting the need for further evidence and potential reclassification of these variants&#46;</p></span><span id="sec0080" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0120">Conclusions</span><p id="par0180" class="elsevierStylePara elsevierViewall">This work presents the preliminary results obtained from a small population&#46; These findings aim to generate new hypotheses and lines of research&#46; Our results suggest that combining CTC characterization with the molecular profiling of the tumor might elucidate tumor heterogeneity&#46; A more extensive study with a larger population would enable the identification and stratification of resectable NSCLC patients at higher risk of recurrence&#46;</p></span><span id="sec0085" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0125">Funding</span><p id="par0185" class="elsevierStylePara elsevierViewall">This work was supported by the Health Counseling from the Andalusian <span class="elsevierStyleGrantSponsor" id="gs1">Government and GlaxoSmithKline Spain &#40;GSK Spain&#41;</span> &#40;grant number&#58; <span class="elsevierStyleGrantNumber" refid="gs1">PIP-0192-2020</span>&#41;&#46;</p></span><span id="sec0090" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0130">Authors&#8217; Contributions</span><p id="par0190" class="elsevierStylePara elsevierViewall">M&#46;J&#46;S&#46; conceptualized the study and acquired the funding&#46; M&#46;J&#46;S&#46; and M&#46;dC&#46;G&#46;N&#46; leaded the study&#46; A&#46;G&#46;D&#46;&#44; M&#46;J&#46;S&#46;&#44; M&#46;dC&#46;G&#46;N&#46; designed the experiments&#46; D&#46;dM&#46;P&#46; and A&#46;G&#46;D&#46; processed samples from patients&#46; A&#46;G&#46;D&#46; and M&#46;J&#46;M&#46; carried out the experiments&#46; M&#46;J&#46;M&#46; and C&#46;B&#46;L&#46; recruited the patients involved in the study&#46; J&#46;E&#46;H&#46; supervised clinical management&#46; F&#46;O&#46;&#44; A&#46;G&#46;D&#46; and A&#46;G&#46;M&#46; performed bioinformatic analyses&#46; A&#46;G&#46;D&#46; visualized the data&#46; A&#46;G&#46;D&#46; and M&#46;J&#46;M&#46; wrote the manuscript&#46; J&#46;L&#46;H&#46; assessed the diagnostic of the different histological subtypes of the patients&#46; M&#46;J&#46;S&#46;&#44; M&#46;dC&#46;G&#46;N&#46;&#44; D&#46;dM&#46;P&#46;&#44; D&#46;L&#46;&#44; J&#46;A&#46;L&#46;&#44; C&#46;B&#46;L&#46;&#44; B&#46;A&#46;N&#44; P&#46;J&#46;R&#46; and C&#46;R&#46; conducted manuscript review and editing&#46; All authors have read and agreed to the published version of the manuscript&#46;</p></span><span id="sec0095" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0135">Conflict of Interest</span><p id="par0195" class="elsevierStylePara elsevierViewall">The authors have no competing interests to declare that are relevant to the content of this article&#46;</p></span><span id="sec0100" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0140">Data Availability Statement</span><p id="par0200" class="elsevierStylePara elsevierViewall">Variants found in this work are accessible through ClinVar accession numbers SCV003806285 and SCV003806424&#46;</p></span></span>
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              "titulo" => "Introduction"
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          "titulo" => "Keywords"
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          "titulo" => "Abbreviations"
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          "identificador" => "sec0005"
          "titulo" => "Introduction"
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          "identificador" => "sec0010"
          "titulo" => "Material and Methods"
          "secciones" => array:7 [
            0 => array:2 [
              "identificador" => "sec0015"
              "titulo" => "Study Design and Patients"
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            1 => array:2 [
              "identificador" => "sec0020"
              "titulo" => "gDNA extraction from tissue samples and quantification"
            ]
            2 => array:2 [
              "identificador" => "sec0025"
              "titulo" => "Library Preparation"
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            3 => array:2 [
              "identificador" => "sec0030"
              "titulo" => "Sequencing Data Analysis"
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            4 => array:2 [
              "identificador" => "sec0035"
              "titulo" => "Collection of Blood Samples&#46; Enrichment and Isolation of CTCs"
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              "identificador" => "sec0040"
              "titulo" => "CTC Characterization by Immunofluorescence"
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              "titulo" => "Statistical Analysis"
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          "titulo" => "Results"
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            0 => array:2 [
              "identificador" => "sec0055"
              "titulo" => "Detection of Somatic Variants in gDNA From Matched Tumoral&#8211;Peritumoral Tissue Biopsies"
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            1 => array:2 [
              "identificador" => "sec0060"
              "titulo" => "Analyze Mutually Exclusive and Co-occurring Events"
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              "identificador" => "sec0065"
              "titulo" => "CTCs&#58; Isolation&#44; Characterization&#44; and Prognostic Role"
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              "identificador" => "sec0070"
              "titulo" => "Integrative Analysis of Tissue Molecular Profiles and Presence of CTCs"
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    "pdfFichero" => "main.pdf"
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    "fechaRecibido" => "2024-05-07"
    "fechaAceptado" => "2024-08-20"
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        0 => array:4 [
          "clase" => "keyword"
          "titulo" => "Keywords"
          "identificador" => "xpalclavsec1893421"
          "palabras" => array:6 [
            0 => "NSCLC"
            1 => "NGS"
            2 => "CTCs"
            3 => "EZH2"
            4 => "MRD"
            5 => "Liquid biopsy"
          ]
        ]
        1 => array:4 [
          "clase" => "abr"
          "titulo" => "Abbreviations"
          "identificador" => "xpalclavsec1893422"
          "palabras" => array:12 [
            0 => "CK"
            1 => "COPD"
            2 => "CTCs"
            3 => "gDNA"
            4 => "IQR"
            5 => "MRD"
            6 => "NGS"
            7 => "NSCLC"
            8 => "OS"
            9 => "PFS"
            10 => "SEM"
            11 => "VUS"
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        "titulo" => "Abstract"
        "resumen" => "<span id="abst0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0015">Introduction</span><p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">Non-small cell lung cancer &#40;NSCLC&#41; is the most common type of lung neoplasm&#46; Despite surgical resection&#44; it has a high relapse rate&#44; accounting for 30&#8211;55&#37; of all cases&#46; Next-generation sequencing &#40;NGS&#41; based on a customized gene panel and the analysis of circulating tumor cells &#40;CTCs&#41; can help identify heterogeneity&#44; stratify high-risk patients&#44; and guide treatment decisions&#46; In this descriptive study involving a small prospective cohort&#44; we focus on the phenotypic characterization of CTCs&#44; particularly concerning EZH2 expression &#40;a member of the Polycomb Repression Complex 2&#41;&#44; as well as on the mutation profiles of the tissue using a customized gene panel and their association with poor outcomes in NSCLC&#46;</p></span> <span id="abst0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0020">Methods</span><p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">Isolation and characterization of EZH2 on CTCs were evaluated before surgical resection &#40;CTC1&#41; and one month after surgery &#40;CTC2&#41; in resectable NSCLC patients&#46; Targeted NGS was performed using a customized 50-gene panel on tissue samples from a subset of patients&#46;</p></span> <span id="abst0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0025">Results</span><p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">76 patients with resectable NSCLC were recruited&#46; The top mutated genes in the cohort included <span class="elsevierStyleItalic">TP53</span>&#44; <span class="elsevierStyleItalic">FLT1</span>&#44; <span class="elsevierStyleItalic">MUC5AC</span>&#44; <span class="elsevierStyleItalic">EGFR</span>&#44; and <span class="elsevierStyleItalic">NLRP3</span>&#46; Pair of genes that had mutually exclusive mutations was <span class="elsevierStyleItalic">TP53-RIN3</span>&#44; and pairs of genes with co-occurring mutations were <span class="elsevierStyleItalic">CD163-TLR4</span>&#44; <span class="elsevierStyleItalic">FGF10-FOXP2</span>&#44; <span class="elsevierStyleItalic">ADAMTSL3-FLT1</span>&#44; <span class="elsevierStyleItalic">ADAMTSL3-MUC5AC</span> and <span class="elsevierStyleItalic">MUC5AC-NLRP3</span>&#46; CTCs decreased significantly between the two time points CTC1 and CTC2 &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;0001&#41;&#44; and CTCs&#43; patients with high EZH2 expression had an 87&#37; increased risk of death &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;018&#41;&#46;</p></span> <span id="abst0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0030">Conclusions</span><p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">Integrating molecular profiling of tumors and CTC characterization can provide valuable insights into tumor heterogeneity and improve patient stratification for resectable NSCLC&#46;</p></span>"
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            "titulo" => "Methods"
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          3 => array:2 [
            "identificador" => "abst0025"
            "titulo" => "Conclusions"
          ]
        ]
      ]
    ]
    "NotaPie" => array:1 [
      0 => array:3 [
        "etiqueta" => "1"
        "nota" => "<p class="elsevierStyleNotepara" id="npar0015">Contributed equally&#46;</p>"
        "identificador" => "fn1"
      ]
    ]
    "apendice" => array:1 [
      0 => array:1 [
        "seccion" => array:1 [
          0 => array:4 [
            "apendice" => "<p id="par0215" class="elsevierStylePara elsevierViewall">The followings are the supplementary data to this article&#58;<elsevierMultimedia ident="upi0005"></elsevierMultimedia></p>"
            "etiqueta" => "Appendix A"
            "titulo" => "Supplementary Data"
            "identificador" => "sec0110"
          ]
        ]
      ]
    ]
    "multimedia" => array:9 [
      0 => array:7 [
        "identificador" => "fig0005"
        "etiqueta" => "Fig&#46; 1"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
            "imagen" => "gr1.jpeg"
            "Alto" => 5612
            "Ancho" => 3483
            "Tamanyo" => 1161891
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        ]
        "descripcion" => array:1 [
          "en" => "<p id="spar0030" class="elsevierStyleSimplePara elsevierViewall">Mutational landscape of the sequenced patients&#46; &#40;A&#41; Descriptive flow of all the variants found in the 23 patients &#40;Sankey plot&#41; summarizing all the characteristics&#46; &#40;B&#41; Oncoplot summarizing mutational profile for each patient and gene&#46; Top bar plot represents the number of mutations for each patient filled depending on the consequence of mutation&#46; Right side bar plot illustrates the number of samples affected by mutation in the given gene&#44; filled depending on the consequence of mutation&#46; Clinical stage of each patient is also present below sample names&#44; with corresponding proportions of transitions&#47;transversions&#46;</p>"
        ]
      ]
      1 => array:7 [
        "identificador" => "fig0010"
        "etiqueta" => "Fig&#46; 2"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
            "imagen" => "gr2.jpeg"
            "Alto" => 2580
            "Ancho" => 3733
            "Tamanyo" => 747720
          ]
        ]
        "descripcion" => array:1 [
          "en" => "<p id="spar0035" class="elsevierStyleSimplePara elsevierViewall">Somatic interactions between pairs of genes&#46; Green color is for co-occurrence and pink for mutually exclusive&#44; in boxes&#46; Purple genes are tumor genes&#44; red are inflammation genes&#44; green are COPD-associated genes and black are lung development genes&#46; Number in brackets represents the count of mutations for a given gene&#46; &#42;&#58; <span class="elsevierStyleItalic">p</span>-Value<span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05 for Fisher&#39;s exact test&#46;</p>"
        ]
      ]
      2 => array:7 [
        "identificador" => "fig0015"
        "etiqueta" => "Fig&#46; 3"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
            "imagen" => "gr3.jpeg"
            "Alto" => 935
            "Ancho" => 1500
            "Tamanyo" => 64396
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        ]
        "descripcion" => array:1 [
          "en" => "<p id="spar0040" class="elsevierStyleSimplePara elsevierViewall">Differences in the number of variants between no recurrence vs&#46; recurrence patients&#46; Number of variants presented in both types of patients&#44; grouped by clinical significance&#46; P<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>VUS&#58; Pathogenic variants and VUS&#44; VUS<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>B&#58; VUS and benign variants&#44; P<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>B&#58; Pathogenic variants and benign variants&#44; Total&#58; all types of variants&#46; &#42;&#58; <span class="elsevierStyleItalic">p</span>-value<span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#44; ns is for not significance&#46; Mean with SEM is represented&#46; <span class="elsevierStyleItalic">t</span> test was applied&#46;</p>"
        ]
      ]
      3 => array:7 [
        "identificador" => "fig0020"
        "etiqueta" => "Fig&#46; 4"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
            "imagen" => "gr4.jpeg"
            "Alto" => 1002
            "Ancho" => 1530
            "Tamanyo" => 76697
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        ]
        "descripcion" => array:1 [
          "en" => "<p id="spar0045" class="elsevierStyleSimplePara elsevierViewall">Dynamics of circulating tumor cells &#40;CTCs&#41; and expression of EZH2 in CTCs from NSCLC patients&#46; &#40;A&#41; Differences in the number of CTCs between CTC1 &#40;before surgery&#41; and CTC2 &#40;one month after surgery&#41;&#46; &#42;&#42;&#42;&#42;&#58; <span class="elsevierStyleItalic">p</span> value for Wilcoxon test&#44; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;0001&#46; &#40;B&#41; Comparison of the percentage of expression of EZH2 at CTC1 and CTC2&#46; Abbreviation n&#46;s&#46; is not significant for Mann&#8211;Whitney <span class="elsevierStyleItalic">U</span> test&#44; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;2562&#41;&#46;</p>"
        ]
      ]
      4 => array:7 [
        "identificador" => "fig0025"
        "etiqueta" => "Fig&#46; 5"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
            "imagen" => "gr5.jpeg"
            "Alto" => 3191
            "Ancho" => 3417
            "Tamanyo" => 391667
          ]
        ]
        "descripcion" => array:1 [
          "en" => "<p id="spar0050" class="elsevierStyleSimplePara elsevierViewall">Prognostic role of CTCs and EZH2 expression on CTCs&#46; Kaplan&#8211;Meier at CTC1 of the effect of the number of CTCs based on a cut-off &#40;high<span class="elsevierStyleHsp" style=""></span>&#8805;<span class="elsevierStyleHsp" style=""></span>2&#47;low<span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>2&#41; in the relapse-free survival &#40;<span class="elsevierStyleBold">A</span>&#41; and overall survival &#40;B&#41;&#46; Kaplan&#8211;Meier of the influence of the EZH2 expression cut-off in the progression-free survival &#40;C&#41; and overall survival &#40;D&#41;&#46; Log-Rank &#40;Mantel&#8211;Cox&#41; test was applied in all the panels&#46;</p>"
        ]
      ]
      5 => array:8 [
        "identificador" => "tbl0005"
        "etiqueta" => "Table 1"
        "tipo" => "MULTIMEDIATABLA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "detalles" => array:1 [
          0 => array:3 [
            "identificador" => "at1"
            "detalle" => "Table "
            "rol" => "short"
          ]
        ]
        "tabla" => array:2 [
          "leyenda" => "<p id="spar0060" class="elsevierStyleSimplePara elsevierViewall">The annotation of these variants and the clinical information of the patients that contained them are presented&#46; HGVS&#58; Human Genome Variation Society nomenclature&#44; ID&#58; identifier for the variant&#44; AF&#58; allele frequency&#44; ACMG&#58; The American College of Medical Genetics and Genomics classification&#44; COPD&#58; Chronic Obstructive Pulmonary Disease&#44; P&#58; pathogenic&#44; LP&#58; likely pathogenic&#44; VUS&#58; variants of uncertain significance&#44; F&#58; female&#44; M&#58; male&#44; ADC&#58; adenocarcinoma&#44; SCC&#58; squamous-cell carcinoma&#46;</p>"
          "tablatextoimagen" => array:1 [
            0 => array:2 [
              "tabla" => array:1 [
                0 => """
                  <table border="0" frame="\n
                  \t\t\t\t\tvoid\n
                  \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Gene&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Transcript&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">HGVS&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">ID&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">AF&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Clinical Significance&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">ACMG Criteria&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Sample&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Gender&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Histology&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Lymph Node Status&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Stage&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">COPD&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Recurrence&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Vital Status&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Cause of Death&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">TP53</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;000546&#46;6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;527G&#62;A&#40;p&#46;Cys176Tyr&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">rs786202962&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;192&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">P&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PM1&#44; PM2&#44; PM5&#44; PP2&#44; PP3&#44; PP5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PCT68&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">F&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">SCC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">N0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">I&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Yes&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Alive&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">TP53</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;000546&#46;6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;584T&#62;C&#40;p&#46;Ile195Thr&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">rs760043106&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;15&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">P&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PM1&#44; PM2&#44; PM5&#44; PP3&#44; PP5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PCT80&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">F&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">ADC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">N0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">I&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Yes&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Alive&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">TP53</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;000546&#46;6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;469G&#62;T&#40;p&#46;Val157Phe&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">rs121912654&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;465&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">VUS&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PM1&#44; PM2&#44; PP3&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PCT33&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">SCC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">N2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">III&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Yes&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Deceased&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Other&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">RB1</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;000321&#46;3&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;1172C&#62;G&#40;p&#46;Ser391&#42;&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">COSV57303024&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;374&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">LP&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PVS1&#44; PM2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">FAM13A</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;014883&#46;4&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;2465C&#62;T&#40;p&#46;Pro822Leu&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">rs377446507&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;174&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">VUS&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PP3&#44; PM2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PCT65&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">SCC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">N1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">II&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Alive&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">RB1</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;000321&#46;3&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;380&#43;2T&#62;C&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">COSV57310513&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;006789&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">LP&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PVS1&#44; PM2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PCT76&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">ADC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">N1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">II&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Alive&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">TP53</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;000546&#46;6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;314G&#62;T&#40;p&#46;Gly105Val&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">rs587781504&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;079&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">LP&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PM1&#44; PM2&#44; PM5&#44; PP2&#44; PP3&#44; PP5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">TP53</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;000546&#46;6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;818G&#62;A&#40;p&#46;Arg273His&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">rs28934576&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;009466&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">P&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PS3&#44; PS4&#44; PM1&#44; PM2&#44; PM5&#44; PP3&#44; PP5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PCT45&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">ADC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">N0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">III&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Alive&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">EGFR</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;005228&#46;5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;2235&#95;2249del&#40;p&#46;Glu746&#95;Ala750del&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">rs121913421&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;081&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">P&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PS1&#44; PM4&#44; PM1&#44; PM2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PCT46&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">F&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">ADC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">N1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">III&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Deceased&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Other&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">TP53</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;000546&#46;6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;622G&#62;T&#40;p&#46;Asp208Tyr&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">COSV52919029&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;031&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">LP&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PM1&#44; PM2&#44; PP3&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PCT47&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">ADC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">N1<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>N2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">III&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Yes&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Alive&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">EGFR</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;005228&#46;5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;2573T&#62;G&#40;p&#46;Leu858Arg&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">rs121434568&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;109&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">P&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PM1&#44; PM2&#44; PM5&#44; PP3&#44; PS1&#44; PP5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PCT74&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">F&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">ADC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">N1<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>N2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">III&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Yes&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Yes&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Deceased&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Tumor progression&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t">rs1057519990&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t  " align="left" valign="\n
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                  """
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                  \t\t\t\t  " colspan="5" align="left" valign="\n
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                  \t\t\t\t">&#40;<a class="elsevierStyleCrossRef" href="#tblfn0010">&#42;&#42;</a>&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Mean &#40;SD&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Non-smoker&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">9 &#40;64&#46;3&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Smoker&#47;former smoker&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleItalic">PET &#40;SUVmax&#41;</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">n&#46;s&#46;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Median &#40;<span class="elsevierStyleItalic">Q</span>1&#44; <span class="elsevierStyleItalic">Q</span>3&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">7&#46;69 &#40;2&#46;72&#44; 11&#46;78&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">6&#46;04 &#40;3&#46;67&#44; 10&#46;71&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Range&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleItalic">Resection</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">0&#46;796&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Sublobar resection&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">3 &#40;60&#46;0&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">2 &#40;40&#46;0&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Segmentectomy&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">2 &#40;100&#46;0&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">0 &#40;0&#46;0&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Lobectomy&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">37 &#40;62&#46;7&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">22 &#40;37&#46;3&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Bilobectomy&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">5 &#40;83&#46;3&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">1 &#40;16&#46;7&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Pneumonectomy&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Chemo<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>radiotherapy&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Yes&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Distant&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">3 &#40;42&#46;9&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-entry\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">Vital status</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
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                  \t\t\t\t">0&#46;801&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">n&#46;s&#46;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-entry\n
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                  \t\t\t\t\ttop\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">36 &#40;66&#46;7&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">18 &#40;33&#46;3&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Deceased&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">14 &#40;63&#46;6&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">8 &#40;36&#46;4&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-entry\n
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                  \t\t\t\t\ttop\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">Cause of death</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="" valign="\n
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                  \t\t\t\t">0&#46;872&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-entry\n
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                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Tumor progression&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">4 &#40;57&#46;1&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">3 &#40;42&#46;8&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">8 &#40;66&#46;7&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">4 &#40;33&#46;3&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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Original Article
Resectable Non-Small Cell Lung Cancer Heterogeneity and Recurrence Assessed by Tissue Next-Generation Sequencing Genotyping and Circulating Tumor Cell EZH2 Characterization
Abel Garcia-Diaza,1, María José Moyano-Rodríguezb,1, María del Carmen Garrido-Navasa, Diego de Miguel-Perezc, Jose Expósito-Hernándezd,e, Bernardino Alcázar-Navarretef, Francisco Ortuñog, David Landeiraa, Pedro J. Romeroh, Adrian Garcia-Morenoa, Jose A. Lorentea, Javier Lopez-Hidalgod,i, Clara Bayarri-Larab,
Corresponding author
ci.bayarri@gmail.com

Corresponding authors.
, Maria Jose Serranoa,d,i,
Corresponding author
mjose.serrano@genyo.es

Corresponding authors.
a GENYO Centre for Genomics and Oncological Research: Pfizer, University of Granada, Andalusian Regional Government, Liquid Biopsy and Cancer Interception Group, 18016 Granada, Spain
b Department of Thoracic Surgery, Virgen de las Nieves University Hospital, Av de las Fuerzas Armadas, 2, 18014 Granada, Spain
c Center for Thoracic Oncology, Tisch Cancer Institute, Mount Sinai Medical System & Icahn School of Medicine, Mount Sinai, New York, NY, USA
d IBS Granada, Instituto de Investigacion Biosanitaria de Granada, 18012 Granada, Spain
e Comprehensive Oncology Division, Virgen de las Nieves University Hospital, Av de las Fuerzas Armadas, 2, 18014 Granada, Spain
f Department of Pneumology Virgen de las Nieves University Hospital, Av de las Fuerzas Armadas, 2, 18014 Granada, Spain
g Department of Computer Engineering, Automation and Robotics, University of Granada, Granada 18071, Spain
h Department of Medicine School of Medicine, University of Granada, 18016 Granada, Spain
i Molecular Pathology Lab, Pathology Service, Virgen de las Nieves University Hospital, Av de las Fuerzas Armadas, 2, 18014 Granada, Spain
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      <span class="elsevierStyleSections"><span id="sec0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0045">Introduction</span><p id="par0005" class="elsevierStylePara elsevierViewall">Lung cancer&#44; which caused 1&#46;8 million deaths in 2022&#44;<a class="elsevierStyleCrossRef" href="#bib0160"><span class="elsevierStyleSup">1</span></a> is mainly represented by non-small cell lung cancer &#40;NSCLC&#41;&#44; which makes up 85&#37; of cases and has a 5-year survival rate of 20&#37; that varies according to tumor stages&#46;<a class="elsevierStyleCrossRef" href="#bib0165"><span class="elsevierStyleSup">2</span></a> Factors that increase the risk of NSCLC include smoking&#44; environmental exposures&#44; and genetic expression dysregulation due to mutations in genes like <span class="elsevierStyleItalic">EGFR</span>&#44; <span class="elsevierStyleItalic">KRAS</span>&#44; and <span class="elsevierStyleItalic">TP53</span>&#44; among others&#46;<a class="elsevierStyleCrossRef" href="#bib0170"><span class="elsevierStyleSup">3</span></a> NSCLC is mainly histologically divided into adenocarcinoma and squamous cell carcinoma&#46; According to the last European Society for Medical Oncology &#40;ESMO&#41; guidelines surgery remains the mainstay treatment for stages I&#8211;II and resectable stage III NSCLC&#46; After surgery&#44; adjuvant chemotherapy should be offered to patients with resected stages IIB and III NSCLC and can be considered in patients with stage IIA&#46;<a class="elsevierStyleCrossRef" href="#bib0175"><span class="elsevierStyleSup">4</span></a> Management of locally advanced stage &#40;IIIA&#47;B&#41; NSCLC consists of multimodal treatment including surgery&#44; chemotherapy&#44; chemoradiotherapy or immune checkpoint inhibitors&#47;tyrosine kinase inhibitors&#44; depending on disease and patient status and the molecular tumor profile&#46;<a class="elsevierStyleCrossRefs" href="#bib0180"><span class="elsevierStyleSup">5&#44;6</span></a> Despite the treatments used&#44; 30&#8211;35&#37; of patients with NSCLC relapse within five years&#44; mainly due to minimal residual disease &#40;MRD&#41; from resection and high genetic heterogeneity&#46;<a class="elsevierStyleCrossRef" href="#bib0170"><span class="elsevierStyleSup">3</span></a> Tumors harbor driver mutations &#40;i&#46;e&#46; <span class="elsevierStyleItalic">EGFR</span>&#44; <span class="elsevierStyleItalic">TP53</span>&#41; and branch mutations that cause intratumorally heterogeneity&#44; being responsible for tumor resistance and survival&#46;<a class="elsevierStyleCrossRefs" href="#bib0170"><span class="elsevierStyleSup">3&#44;7</span></a> Precision medicine uses next-generation sequencing &#40;NGS&#41; to identify heterogeneity&#44; select treatments&#44; and unveil the origin and course of disease&#46;<a class="elsevierStyleCrossRefs" href="#bib0195"><span class="elsevierStyleSup">8&#44;9</span></a> Nevertheless&#44; among various strategies in NGS&#44; gene panels distinguish themselves for their cost-effectiveness and precision&#46;<a class="elsevierStyleCrossRefs" href="#bib0205"><span class="elsevierStyleSup">10&#44;11</span></a></p><p id="par0010" class="elsevierStylePara elsevierViewall">Liquid biopsy &#40;LB&#41; monitors the real-time evolution of cancer and circulating tumor cells &#40;CTCs&#41; are one of the elements that correlate with MRD and the risk of relapse in lung cancer patients undergoing surgery&#46;<a class="elsevierStyleCrossRefs" href="#bib0215"><span class="elsevierStyleSup">12&#44;13</span></a> However&#44; the phenotypic diversity of CTCs in lung cancer requires a clinically meaningful profile&#46;<a class="elsevierStyleCrossRefs" href="#bib0220"><span class="elsevierStyleSup">13&#44;14</span></a> CTCs reflect the complexity of tissues&#44; and it is possible to identify clones of resistant tumors that cause relapse&#46; The epithelial&#8211;mesenchymal transition marks the presence of diverse phenotypes and metastatic potential regulated by EZH2&#44; associated with NSCLC progression&#44; metastasis&#44; and maintenance of cancer cell characteristics&#46;<a class="elsevierStyleCrossRefs" href="#bib0230"><span class="elsevierStyleSup">15&#44;16</span></a> Screening for CTC&#8722; EZH2&#43; could help stratify the risk of relapse&#47;death and guide treatments&#46;</p><p id="par0015" class="elsevierStylePara elsevierViewall">The primary objective of this study is to investigate the relationship between the molecular profiles of resectable NSCLC tissue and the presence of CTCs that express EZH2 subpopulations&#46; We aim to determine if these molecular profiles and CTC phenotypes can predict clinical outcomes and the likelihood of relapse&#46; Specifically&#44; we hypothesize that certain molecular characteristics are associated with an increased probability of detecting CTCs&#44; which may serve as minimal residual disease &#40;MRD&#41; indicators&#46; Our results will explore these associations to provide clearer insights into how molecular profiles and CTC presence can inform patient prognosis and treatment strategies&#46;</p></span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0050">Material and Methods</span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0055">Study Design and Patients</span><p id="par0020" class="elsevierStylePara elsevierViewall">Conducted between July 2017 and December 2019&#44; this prospective observational cohort study focused on NSCLC patients undergoing surgical treatment &#40;I&#8211;III&#41; at a single tertiary hospital&#46; Surgery consisted of anatomical pulmonary resection and&#44; systematic lymph node dissection for clinical stages II and IIIA and lobe-specific lymph node dissection for stage I&#46; For the LB analyzes&#44; the peripheral blood samples were collected at baseline &#40;CTC1&#41; and one month after surgery &#40;CTC2&#41;&#46; Tumor tissue was extracted during surgery&#44; tumoral and peritumoral samples were obtained through a sterile field and placed in RNA Later tubes &#40;AM7020&#44; ThermoFisher Scientific&#44; Waltham&#44; Massachusetts&#44; USA&#41; for preservation&#46;</p><p id="par0025" class="elsevierStylePara elsevierViewall">The pathologic stage was determined by the 8th edition of the international tumor-node-metastasis &#40;TNM&#41; system and&#44; according to the World Health Organization classification&#44; the histologic subtypes of lung cancer were assigned&#46; After surgery&#44; patients were discussed in the hospital&#39;s thoracic multidisciplinary oncology committee&#46; Adjuvant chemotherapy was indicated in stages IIb and III following the ESMO guidelines&#44; and it was considered in patients with stage IIA&#46;</p><p id="par0030" class="elsevierStylePara elsevierViewall">The follow-up protocol consisted of surveillance every 6 months for the first 3 years&#44; which included a chest and abdominal CT scan&#44; with optional FDG-PET if required&#46; Then&#44; the follow-up was yearly&#46; Clinical outcomes regarding overall survival &#40;OS&#41; and progression-free survival &#40;PFS&#41; were estimated using the Kaplan&#8211;Meier method and both were calculated at 5 years after surgical treatment&#46;</p><p id="par0035" class="elsevierStylePara elsevierViewall">The study&#44; adhering to the Declaration of Helsinki and approved by regional institutional review boards&#44; obtained written informed consent from all participants before enrollment&#46;</p></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0060">gDNA extraction from tissue samples and quantification</span><p id="par0040" class="elsevierStylePara elsevierViewall">Matched tumoral&#47;peritumoral tissue biopsies were collected from patients that underwent surgical treatment&#46; gDNA was extracted using the NucleoSpin&#174; Tissue kit &#40;740952&#46;50&#44; Macherey-Nagel&#44; D&#252;ren&#44; Germany&#41;&#44; following the manufacturer&#39;s instructions&#46; Briefly&#44; 25<span class="elsevierStyleHsp" style=""></span>mg of tissue was used for the extraction&#46; Tissues were homogenized using the TissueLyser LT instrument &#40;Qiagen&#44; Venlo&#44; Netherlands&#41; for 5<span class="elsevierStyleHsp" style=""></span>min at 50<span class="elsevierStyleHsp" style=""></span>oscillations&#47;s&#46; After that&#44; samples were lysed with Proteinase K solution at a final concentration of 2&#46;7<span class="elsevierStyleHsp" style=""></span>mg&#47;ml at 56<span class="elsevierStyleHsp" style=""></span>&#176;C overnight in a shaker&#46; After purification and precipitation&#44; DNA was bound to a silica column&#44; washed twice&#44; and the column was dried out&#46; DNA was eluted in a final volume of 75<span class="elsevierStyleHsp" style=""></span>&#956;l&#46; Concentration was measured using a NanoDrop&#8482; 2000 instrument &#40;ThermoFisher&#41; at a wavelength of 260<span class="elsevierStyleHsp" style=""></span>nm&#46;</p></span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0065">Library Preparation</span><p id="par0045" class="elsevierStylePara elsevierViewall">Sequencing libraries were constructed using the KAPA HyperCap workflow v3&#46;0 &#40;Roche&#44; Basel&#44; Switzerland&#41; with a customized gene panel&#44; following the manufacturer&#39;s protocol&#46; This customized panel was developed to cover 50 genes &#40;exons&#41; involved in four different biological processes&#58; tumor promotion &#40;<span class="elsevierStyleItalic">ROS1</span>&#44; <span class="elsevierStyleItalic">ALK</span>&#44; <span class="elsevierStyleItalic">TP53</span>&#44; <span class="elsevierStyleItalic">APC</span>&#44; <span class="elsevierStyleItalic">RB1</span>&#44; <span class="elsevierStyleItalic">RET</span>&#44; <span class="elsevierStyleItalic">FLT1</span>&#44; <span class="elsevierStyleItalic">PTEN</span>&#44; <span class="elsevierStyleItalic">STK11</span>&#44; <span class="elsevierStyleItalic">TET2</span>&#44; <span class="elsevierStyleItalic">CTNNB1</span>&#44; <span class="elsevierStyleItalic">MLH1</span>&#44; <span class="elsevierStyleItalic">EGFR</span>&#44; <span class="elsevierStyleItalic">DDR2</span>&#44; <span class="elsevierStyleItalic">SLC22A18</span>&#44; <span class="elsevierStyleItalic">AKT1</span>&#41;&#44; Chronic Obstructive Pulmonary Disease &#40;COPD&#41; associated genes &#40;<span class="elsevierStyleItalic">ADAMTSL3</span>&#44; <span class="elsevierStyleItalic">ADAM19</span>&#44; <span class="elsevierStyleItalic">PTCH1</span>&#44; <span class="elsevierStyleItalic">ADGRG6</span>&#44; <span class="elsevierStyleItalic">HTR4</span>&#44; <span class="elsevierStyleItalic">FAM13A</span>&#44; <span class="elsevierStyleItalic">CHRNB4</span>&#44; <span class="elsevierStyleItalic">RIN3</span>&#44; <span class="elsevierStyleItalic">TGFB2</span>&#44; <span class="elsevierStyleItalic">IREB2</span>&#44; <span class="elsevierStyleItalic">SERPINA1</span>&#44; <span class="elsevierStyleItalic">CHRNA3</span>&#44; <span class="elsevierStyleItalic">STN1</span>&#44; <span class="elsevierStyleItalic">CHRNA5</span>&#44; <span class="elsevierStyleItalic">AGER</span>&#41;&#44; lung development &#40;<span class="elsevierStyleItalic">FGF10</span>&#44; <span class="elsevierStyleItalic">FOXA1</span>&#44; <span class="elsevierStyleItalic">FOXA2</span>&#44; <span class="elsevierStyleItalic">FOXP2</span>&#44; <span class="elsevierStyleItalic">FOXP4</span>&#44; <span class="elsevierStyleItalic">HOXA2</span>&#44; <span class="elsevierStyleItalic">HOXA5</span>&#44; <span class="elsevierStyleItalic">HOXB2</span>&#44; <span class="elsevierStyleItalic">PITX2</span>&#44; <span class="elsevierStyleItalic">WNT2</span>&#44; <span class="elsevierStyleItalic">ALX1</span>&#44; <span class="elsevierStyleItalic">NKX2-1</span>&#41; and inflammation &#40;<span class="elsevierStyleItalic">CD163</span>&#44; <span class="elsevierStyleItalic">S100A4</span>&#44; <span class="elsevierStyleItalic">NLRP3</span>&#44; <span class="elsevierStyleItalic">MUC5AC</span>&#44; <span class="elsevierStyleItalic">EGR1</span>&#44; <span class="elsevierStyleItalic">TLR4</span>&#44; <span class="elsevierStyleItalic">FPR2</span>&#41;&#44; spanning approximately 140<span class="elsevierStyleHsp" style=""></span>kb&#46; It was tested on I&#8211;IIIB stage NSCLC to elucidate the contribution of non-tumoral gene mutations on cancer disease development&#44; intercepting cancer&#46; Libraries were quantified with Qubit 4&#46;0 Fluorometer using dsDNA HS Assay kit&#46; Individual libraries were diluted and pooled together in equal amounts &#40;by mass&#44; equimolarity&#41; for obtaining equal numbers of sequencing reads &#40;HiSeq X System&#44; Illumina&#44; San Diego&#44; California&#44; USA&#41;&#46;</p></span><span id="sec0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0070">Sequencing Data Analysis</span><p id="par0050" class="elsevierStylePara elsevierViewall">Once FASTQ files were demultiplexed and retrieved from the sequencer&#44; files were ready for data processing&#46; The GATK best practices-based on the Somatic Variant Pipeline &#40;<a href="https://www.clinbioinfosspa.es/content/somatic-variant-pipeline">https&#58;&#47;&#47;www&#46;clinbioinfosspa&#46;es&#47;content&#47;somatic-variant-pipeline</a>&#41; from the Clinical Bioinformatics Area from Fundaci&#243;n Progreso y Salud &#40;Andalusian Govern &#8211; Health and Family Counseling&#41; were followed&#46; Briefly&#44; raw data quality was evaluated using the FastQC tool &#40;v0&#46;11&#46;9&#44; Babraham Bioinformatics&#44; Cambridge&#44; UK&#41;&#46; High quality reads were aligned against the human reference genome &#40;GRCh38&#41; using Burrow&#8211;Wheeler Aligner algorithm &#40;<a href="https://github.com/lh3/bwa">https&#58;&#47;&#47;github&#46;com&#47;lh3&#47;bwa</a>&#41;&#46; After filtering steps&#44; tumor with matched normal mode from Mutect2 pipeline from GATK &#40;<a href="https://gatk.broadinstitute.org/hc/en-us">https&#58;&#47;&#47;gatk&#46;broadinstitute&#46;org&#47;hc&#47;en-us</a>&#41; was run for variant calling&#46; Generated filtered VCFs were annotated using VEP &#40;v99&#44; <a href="https://www.ensembl.org/info/docs/tools/vep/index.html">https&#58;&#47;&#47;www&#46;ensembl&#46;org&#47;info&#47;docs&#47;tools&#47;vep&#47;index&#46;html</a>&#41;&#46; Clinical significance was assessed by following the American College of Medical Genetics &#40;ACMG&#41; criteria&#46; R software &#40;v4&#46;2&#46;1&#41; and RStudio &#40;2022&#46;12&#46;0<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>353 version&#41; were used for file parsing and for plot generation&#46; Maftools<a class="elsevierStyleCrossRef" href="#bib0240"><span class="elsevierStyleSup">17</span></a> was used for oncoplot generation and somatic interactions plot&#46;</p></span><span id="sec0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0075">Collection of Blood Samples&#46; Enrichment and Isolation of CTCs</span><p id="par0055" class="elsevierStylePara elsevierViewall">Peripheral blood &#40;10<span class="elsevierStyleHsp" style=""></span>ml&#41; was collected in EDTA BD Vacutainer&#174; tubes and stored at room temperature for processing within the next 4<span class="elsevierStyleHsp" style=""></span>h&#46; CTCs were isolated by density gradient centrifugation&#44; following a previously described protocol&#46;<a class="elsevierStyleCrossRef" href="#bib0215"><span class="elsevierStyleSup">12</span></a> Briefly&#44; blood samples were centrifuged by density gradient with Ficoll Histopaque&#174;-1119 &#40;density<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>1&#46;119<span class="elsevierStyleHsp" style=""></span>g&#47;ml&#41; &#40;Merck&#44; Darmstadt&#44; Germany&#41; for 45<span class="elsevierStyleHsp" style=""></span>min at 700<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleItalic">g</span> with no break&#46; The buffy coat where CTCs and PBMCs are located was carefully extracted&#46; Isolation of CTCs was performed by positive immunomagnetic selection using the Carcinoma Cell Enrichment kit &#40;130-108-339&#44; Miltenyi Biotec&#44; Bergisch Gladbach&#44; Germany&#41;&#46; This selection is based on a multi-cytokeratin antibody &#40;CK3-6H5&#41;&#44; that recognizes the cytoplasmic cytokeratins 7 and 8&#46; Cellular samples were spun down onto slides that were stained and characterized by immunofluorescence&#46;</p></span><span id="sec0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0080">CTC Characterization by Immunofluorescence</span><p id="par0060" class="elsevierStylePara elsevierViewall">The working area was marked with a hydrophobic pen &#40;Dako&#41;&#46; Each sample was hydrated with PBS 1&#215; for 5<span class="elsevierStyleHsp" style=""></span>min and incubated with anti-cytokeratin-FITC antibody&#44; against cytokeratins 7 and 8 &#40;130-060-301&#44; Miltenyi Biotec&#41;&#44; diluted 1&#58;100 in PBS 1&#215; &#8211; Tween 0&#46;1&#37;<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>BSA 2&#37; &#40;working solution&#41; for 45<span class="elsevierStyleHsp" style=""></span>min in the dark at room temperature&#46; After this point&#44; darkness was always maintained&#46; After this incubation&#44; samples were washed with PBS 1&#215; for 5<span class="elsevierStyleHsp" style=""></span>min&#46; A blockade was performed with goat serum diluted at 1&#215; in working solution for 1<span class="elsevierStyleHsp" style=""></span>h&#46; To detect the EZH2 protein&#44; primary rabbit anti-EZH2 antibody &#40;&#35;5246S&#44; Cell Signaling Technology&#41; was diluted 1&#58;100 in the working solution and was incubated for 45<span class="elsevierStyleHsp" style=""></span>min&#46; Then&#44; samples are washed in PBS 1&#215; for 5<span class="elsevierStyleHsp" style=""></span>min&#46; Secondary goat anti-rabbit IgG &#40;Alexa Fluor 555&#44; ThermoFisher&#41; was diluted 1&#58;200 in the working solution and was incubated for 45<span class="elsevierStyleHsp" style=""></span>min to reveal EZH2 expression&#46; Nuclei were counterstained using Hoechst33342 &#40;Fluxion&#41; at 1&#215;&#46; Slides were mounted using SlowFade&#8482; Diamond Antifade Mountant &#40;S36972&#44; ThermoFisher&#41; for visualization in Zeiss LSM 710 confocal microscope&#46;</p><p id="par0065" class="elsevierStylePara elsevierViewall">Evaluation of EZH2 in CTCs was analyzed after previous evaluation on A549 control cell line &#40;CCL-185&#8482;&#44; ATCC&#44; Manassas&#44; Virginia&#44; USA&#41; for antibody specificity &#40;<a class="elsevierStyleCrossRef" href="#sec0110">Supplementary Fig&#46; 1</a>&#41;&#46; Because of the heterogeneity observed in patients with CTCs&#44; the percentage expression of EZH2 was calculated as the number of EZH2&#43; CTCs divided by the total number of CTCs&#46; This calculation aimed to quantify the heterogeneity of CTCs within patients&#46;</p></span><span id="sec0045" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0085">Statistical Analysis</span><p id="par0070" class="elsevierStylePara elsevierViewall">CTCs were assessed both as a continuous variable &#40;number of CTCs per 10<span class="elsevierStyleHsp" style=""></span>ml of blood&#41; and as a binary variable &#40;presence&#47;absence and a cut-off of &#8805;2 CTCs per 10<span class="elsevierStyleHsp" style=""></span>ml&#41;&#46; Cut-offs to distinguish low&#47;high CTC numbers and EZH2 expression were determined using the Cut-off Finder web application &#40;fit for mixture model&#41;&#46;<a class="elsevierStyleCrossRef" href="#bib0245"><span class="elsevierStyleSup">18</span></a> Bivariate analysis examined the relation between clinical features and CTC presence&#47;absence&#46; Clinical variables were evaluated for normality using the Shapiro&#8211;Wilk test&#46;</p><p id="par0075" class="elsevierStylePara elsevierViewall">The clinical characteristics were evaluated by Chi-squared &#40;Gender&#44; Procedure&#44; Histology&#44; Respiratory history&#44; Stage&#44; Adjuvancy&#44; Recurrence and Vital status&#41;&#44; Fisher&#39;s exact test &#40;Tobacco&#44; Resection&#44; Lymph node status&#44; Type of adjuvancy&#44; Site of recurrence&#44; and Cause of death&#41;&#44; <span class="elsevierStyleItalic">t</span>-test &#40;Age&#41; and Mann&#8211;Whitney <span class="elsevierStyleItalic">U</span> test &#40;PET &#8211; SUVmax&#8211;&#44; Tumor size &#40;cm&#41;&#44; overall survival &#8211; OS and progression-free survival &#8211; PFS&#44; both in months&#41;&#46; Parametric variables are expressed as mean with standard deviation and non-parametric variables as median with interquartile range or down&#47;upper quartile &#40;<span class="elsevierStyleItalic">Q</span>1&#44; <span class="elsevierStyleItalic">Q</span>3&#41;&#46;</p><p id="par0080" class="elsevierStylePara elsevierViewall">Wilcoxon Signed-Rank test compared the median of CTC numbers and EZH2 expression between sampling points&#44; to study the dynamics of these variables&#46; Univariate Kaplan&#8211;Meier &#40;log-rank test&#41; analyzed the influence of CTC and EZH2 cut-offs on PFS and OS&#46; Fisher&#39;s exact test evaluated somatic interactions between gene pairs&#44; while <span class="elsevierStyleItalic">t</span>-tests and Fisher&#39;s exact test were further used to compare mutation profiles concerning recurrence and CTC presence&#44; respectively&#46;</p><p id="par0085" class="elsevierStylePara elsevierViewall">Statistical analyses were performed using IBM SPSS Statistics &#40;version 25&#46;0 for MacOS&#44; IBM Corp&#46;&#44; Armonk&#44; NY&#44; USA&#41;&#44; GraphPad Prism &#40;version 9&#46;0&#46;2 for MacOS&#44; GraphPad software&#44; La Jolla&#44; CA&#44; USA&#41;&#44; and R software &#40;v4&#46;2&#46;1&#41; and RStudio &#40;2023&#46;03&#46;0<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>386 version&#41;&#44; with a <span class="elsevierStyleItalic">p</span>-value<span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05 determining statistical significance&#46;</p></span></span><span id="sec0050" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0090">Results</span><p id="par0090" class="elsevierStylePara elsevierViewall">A total of 76 resectable NSCLC patients were enrolled in the study&#46; A descriptive analysis of the cohort is shown in <a class="elsevierStyleCrossRef" href="#sec0110">Supplementary Table 1</a>&#46; Most patients were males &#40;60&#46;5&#37;&#41; and smokers &#40;81&#46;6&#37;&#41;&#44; with a mean age of 66 years&#46; Lobectomy was the main pulmonary resection performed &#40;77&#46;6&#37;&#41; by Video Assisted Thoracic Surgery &#40;VATS&#41; approach &#40;58&#46;0&#37;&#41;&#46; During the follow-up&#44; 20 patients presented cancer recurrence &#40;26&#46;3&#37;&#41;&#46; The median time of PFS was 21 months&#46; The histological evaluation of NSCLC was conducted using hematoxylin&#8211;eosin staining&#46; The specific histologic types &#40;adenocarcinoma or squamous&#41; were assessed by a pathologist &#40;<a class="elsevierStyleCrossRef" href="#sec0110">Supplementary Fig&#46; 2</a>&#41;&#46;</p><span id="sec0055" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0095">Detection of Somatic Variants in gDNA From Matched Tumoral&#8211;Peritumoral Tissue Biopsies</span><p id="par0095" class="elsevierStylePara elsevierViewall">Out of the total of 76 patients&#44; only 27 were eligible for inclusion in the NGS analyses due to meeting the required quality standards for library preparation&#46; Average concentration of extracted gDNA was 316&#46;53<span class="elsevierStyleHsp" style=""></span>ng&#47;&#956;L &#40;range&#58; 43&#46;85&#8211;946&#46;85<span class="elsevierStyleHsp" style=""></span>ng&#47;&#956;L&#41;&#46; Minimal input ensured for library preparation was 500<span class="elsevierStyleHsp" style=""></span>ng&#46; After library preparation&#44; sequencing yielded a mean coverage of approximately 500&#215; along all the samples&#46; From the 27 patients sequenced&#44; only 4 did not pass variant calling filters and they were removed from subsequent analysis&#46; The total number of variants identified for the 23 remaining patients were 143&#44; with an average of 6 variants per patient &#40;range&#58; 1&#8211;25&#41;&#46; Most of the variants &#40;61&#46;5&#37;&#59; 88&#47;143&#41; were non-coding and SNPs &#40;91&#46;6&#37;&#41;&#44; but only 34&#46;3&#37; &#40;49&#47;143&#41; were known variants&#46; Variants were submitted to ClinVar &#40;Accession numbers&#58; SCV003806285&#8211;SCV003806424&#41;&#46; Regarding the clinical significance of the variants&#44; 9&#46;8&#37; &#40;14&#47;143&#41; were benign&#47;likely benign&#44; 83&#46;2&#37; &#40;119&#47;143&#41; were variants of uncertain significance &#40;VUS&#41; and 7&#46;0&#37; &#40;10&#47;143&#41; were pathogenic&#47;likely pathogenic &#40;<a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>A&#41;&#46; Oncoplot in <a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>B summarizes data for the mutational profiles of each patient and gene&#46; The number of mutations for each patient and the number of samples &#40;patients&#41; affected by mutations in that given gene are represented on both top and right plots&#44; respectively&#46; The top 5 most highly mutated genes were&#58; <span class="elsevierStyleItalic">TP53</span>&#44; <span class="elsevierStyleItalic">FLT1</span>&#44; <span class="elsevierStyleItalic">MUC5AC</span>&#44; <span class="elsevierStyleItalic">EGFR</span> and <span class="elsevierStyleItalic">NLRP3</span>&#44; presented in 43&#37;&#44; 39&#37;&#44; 39&#37;&#44; 26&#37; and 26&#37; of the samples&#44; respectively&#46; Cancer stage&#44; presence&#47;absence of CTCs&#44; expression of EZH2 and mutational signatures &#40;transitions and transversions&#41; are also provided for each patient &#40;<a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>B&#41;&#46;</p><elsevierMultimedia ident="fig0005"></elsevierMultimedia><p id="par0100" class="elsevierStylePara elsevierViewall">Full description of the pathogenic&#44; likely pathogenic variants and hot VUS found is described in <a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#46; Both PCT74 and PCT75 patients recurred and had primary point mutations in <span class="elsevierStyleItalic">EGFR</span> and <span class="elsevierStyleItalic">ROS1</span>&#44; respectively&#44; and both had secondary point mutations in <span class="elsevierStyleItalic">TP53</span> &#40;according to allele frequencies&#41;&#46; Contrarily&#44; patients PCT33 and PCT76&#44; had primary mutations in <span class="elsevierStyleItalic">TP53</span> and secondary mutations in <span class="elsevierStyleItalic">RB1</span>&#44; and they did not have recurrence&#44; suggesting that secondary mutations in <span class="elsevierStyleItalic">TP53</span> might have a stronger impact on disease recurrence&#46;</p><elsevierMultimedia ident="tbl0005"></elsevierMultimedia></span><span id="sec0060" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0100">Analyze Mutually Exclusive and Co-occurring Events</span><p id="par0105" class="elsevierStylePara elsevierViewall">Studying if a pair of genes were mutated simultaneously or if they were mutually exclusive&#44; interactions of the somatic variants were analyzed&#46; The top 5 most significant pairs of genes with co-occurring mutations were&#58; <span class="elsevierStyleItalic">CD163-TLR4</span>&#44; <span class="elsevierStyleItalic">FGF10-FOXP2</span>&#44; <span class="elsevierStyleItalic">ADAMTSL3-FLT1</span>&#44; <span class="elsevierStyleItalic">ADAMTSL3-MUC5AC</span> and <span class="elsevierStyleItalic">MUC5AC-NLRP3</span> with <span class="elsevierStyleItalic">p</span>-values of 0&#46;0056&#44; 0&#46;0118&#44; 0&#46;0142&#44; 0&#46;0144&#44; 0&#46;0183&#44; respectively&#46; There was only one significant pair of genes with mutually exclusive mutations&#58; <span class="elsevierStyleItalic">RIN3-TP53</span> &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;0457&#41;&#46; Interactions among all the genes are represented in <a class="elsevierStyleCrossRef" href="#fig0010">Fig&#46; 2</a>&#46; Detailed data of significant pairs of genes highlighted in <a class="elsevierStyleCrossRef" href="#fig0010">Fig&#46; 2</a> can be found in <a class="elsevierStyleCrossRef" href="#sec0110">Supplementary Table 2</a> with additional information for all the significant pairs of genes&#46;</p><elsevierMultimedia ident="fig0010"></elsevierMultimedia><p id="par0110" class="elsevierStylePara elsevierViewall">Most variants were VUS &#40;83&#46;2&#37;&#41; and as they might be reclassified in the future as either pathogenic&#47;likely pathogenic &#40;P&#41; or benign&#47;likely benign &#40;B&#41;&#44; we studied their impact on recurrence by grouping them with either P or B&#46; Two comparisons were done&#58; P<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>VUS &#40;pathogenic variants and VUS&#41; and VUS<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>B &#40;VUS and benign variants&#41;&#46; P<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>VUS showed significant differences &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;0404&#41; between no-recurrence and recurrence patients&#44; with a mean number of variants of 4&#46;8 &#40;standard error of the mean &#8211; SEM<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;65&#41; in the no-recurrence group versus a mean of 11 &#40;SEM<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>5&#46;54&#41; variants in the recurrence group&#46; On the other hand&#44; VUS<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>B showed no significant differences &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;0597&#41; between no-recurrence &#40;mean of 5&#44; SEM<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;71&#41; and recurrence patients &#40;mean of 11&#44; SEM<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>7&#46;09&#41; &#40;<a class="elsevierStyleCrossRef" href="#fig0015">Fig&#46; 3</a>&#41;&#46;</p><elsevierMultimedia ident="fig0015"></elsevierMultimedia></span><span id="sec0065" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0105">CTCs&#58; Isolation&#44; Characterization&#44; and Prognostic Role</span><p id="par0115" class="elsevierStylePara elsevierViewall">The possible correlation between the clinical&#8211;pathological characteristics of the patients and the presence or absence of CTCs &#40;based on cytokeratin&#44; CK&#44; expression&#41; was investigated before surgery &#40;CTC1&#44; <a class="elsevierStyleCrossRef" href="#tbl0010">Table 2</a>&#41; and one month after surgery &#40;CTC2&#44; <a class="elsevierStyleCrossRef" href="#sec0110">Supplementary Table 3</a>&#41;&#46; The presence of CTC before surgery &#40;CTC1&#41; was significantly correlated with age&#44; lymph node status &#40;N&#41;&#44; tumor stage&#44; and adjuvant treatment&#46; Regarding tumor stage&#44; most patients with stages II and III were CTC&#8722; &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;0300&#41;&#46; Focused on N status&#44; most patients with lymph node metastasis were CTC&#8722; &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;0320&#41;&#46; Consequently&#44; it was observed that most CTC&#8722; patients received adjuvant treatment compared with the CTC&#43; patients &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;0040&#41;&#46;</p><elsevierMultimedia ident="tbl0010"></elsevierMultimedia><p id="par0120" class="elsevierStylePara elsevierViewall">At CTC1&#44; CTCs were detected in 26&#47;76 patients &#40;34&#46;2&#37;&#41; with a median number of 3 in those CTCs&#43; patients &#40;IQR<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>3&#44; range<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>1&#8211;8&#41; meanwhile&#44; at CTC2 the presence of CTCs was detected in only 3&#47;76 patients &#40;3&#46;9&#37;&#41; with a median number of 2 CTCs in those CTCs&#43; patients &#40;IQR<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>3&#44; range<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>1&#8211;4&#41;&#46; In addition&#44; the dynamics of CTCs between the two time points&#44; CTC1 and CTC2&#44; were also analyzed&#46; We observed a significant decrease in the number of CTCs because of the surgery &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;0001&#44; Wilcoxon test&#41; &#40;<a class="elsevierStyleCrossRef" href="#fig0020">Fig&#46; 4</a>A&#41;&#46;</p><elsevierMultimedia ident="fig0020"></elsevierMultimedia><p id="par0125" class="elsevierStylePara elsevierViewall">The expression of EZH2 on CTCs was measured &#40;see CTC characterization by immunofluorescence methods Section&#41; at CTC1 in the 26 CTC&#43; patients&#46; We found that 13&#47;26 patients were EZH2&#43; &#40;50&#46;0&#37;&#41;&#44; with a median percentage of expression of 6&#46;25&#37; &#40;IQR&#58; 54&#46;17&#37;&#44; range&#58; 0&#8211;100&#37;&#41;&#46; At CTC2&#44; EZH2 was detected in 2&#47;3 CTC&#43; patients &#40;66&#46;67&#37;&#41;&#44; with a median percentage of expression of EZH2 of 100&#37; &#40;IQR&#58; 100&#37;&#44; range&#58; 0&#8211;100&#37;&#41;&#46; Expression of EZH2 showed no significant differences between CTC1 and CTC2 &#40;<a class="elsevierStyleCrossRef" href="#fig0020">Fig&#46; 4</a>B&#41;&#46;</p><p id="par0130" class="elsevierStylePara elsevierViewall">The prognostic role of circulating tumor cells &#40;CTCs&#41; for PFS and OS was evaluated using either the presence&#47;absence of CTCs or a cut-off of \n
      &lt;!--&#8805;2 CTCs at CTC1. No significant differences were found for OS or PFS using Kaplan&#8211;Meier analysis for the presence/absence of CTCs (--&gt;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;5430 and <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;9070&#44; respectively&#59; see <a class="elsevierStyleCrossRef" href="#sec0110">Supplementary Fig&#46; 3</a>&#41;&#46; Using the CTC cut-off of \n
      &lt;!--&#8805;2 slightly improved both OS and PFS, but again, no significant differences were observed (--&gt;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;1920 and <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;4380&#44; respectively&#41;&#46; OS was measured at 5 years since surgical treatment&#46;</p><p id="par0135" class="elsevierStylePara elsevierViewall">Interestingly&#44; when evaluating EZH2 expression in the CTCs using a cut-off of &#60;70&#37; or &#8805;70&#37; for the previously calculated percentage of expression &#40;EZH2low&#47;EZH2high&#44; respectively&#41;&#44; significant differences in terms of OS were observed at CTC1 &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;0180&#44; Log-Rank&#44; Mantel&#8211;Cox&#41; with EZH2high patients having an 87&#37; greater risk of death&#46; In terms of PFS no significant differences were found for EZH2 characterization &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;9130&#41; &#40;<a class="elsevierStyleCrossRef" href="#fig0025">Fig&#46; 5</a>C and D&#41;&#46;</p><elsevierMultimedia ident="fig0025"></elsevierMultimedia><p id="par0140" class="elsevierStylePara elsevierViewall">At CTC2 neither presence&#47;absence of CTCs nor CTC2low&#47;CTC2high showed significant differences in terms of PFS or OS &#40;<a class="elsevierStyleCrossRef" href="#sec0110">Supplementary Fig&#46; 4</a>&#41;&#46; At this point&#44; Kaplan&#8211;Meier curves could not be calculated for EZH2low&#47;EZH2high in CTCs&#43; patients due to the very small sample size&#46;</p></span><span id="sec0070" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0110">Integrative Analysis of Tissue Molecular Profiles and Presence of CTCs</span><p id="par0145" class="elsevierStylePara elsevierViewall">Presence or absence of CTCs at baseline was analyzed based on the tissue molecular profiles&#46; While we did not find statistical differences between the presence or absence of CTCs and VUS in the studied genes&#44; we did observe a noteworthy trend&#46; There were two out of three patients without CTCs in the follow-up but with VUS in <span class="elsevierStyleItalic">ADAM19</span> that locally recurred &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;1739&#41;&#46; The only patient with CTCs &#40;both at baseline and the follow-up&#41; and VUS in <span class="elsevierStyleItalic">ADAM19</span> did not recur at time of these analyses &#40;<a class="elsevierStyleCrossRef" href="#sec0110">Supplementary Table 4</a>&#41;&#46;</p></span></span><span id="sec0075" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0115">Discussion</span><p id="par0150" class="elsevierStylePara elsevierViewall">This study examines biological data from different approaches in stage I-IIIB NSCLC patients&#44; evaluating the mutational profile of lung tissue with a customized panel of 50 genes and CTCs&#46; Regarding the prognostic role of CTCs&#44; despite this study including a small population&#44; patients with a high percentage of CTC&#8722; EZH2&#43; showed poorer OS&#46; Our recent results are consistent with previous data associating the expression of the EZH2 protein in breast cancer with metastatic recurrence and worse PFS and OS in this study<a class="elsevierStyleCrossRefs" href="#bib0250"><span class="elsevierStyleSup">19&#8211;21</span></a> and in lung cancer with tumor colonization&#46;<a class="elsevierStyleCrossRef" href="#bib0235"><span class="elsevierStyleSup">16</span></a> Due to the essential and heterogeneous role of EZH2 in regulating various sets of genes in cancer&#44; many clinical trials have emerged to test inhibitors against this target&#46;<a class="elsevierStyleCrossRef" href="#bib0265"><span class="elsevierStyleSup">22</span></a> However&#44; our results also suggest the importance of characterizing the CTCs to identify the aggressiveness of cell subpopulations&#46;<a class="elsevierStyleCrossRefs" href="#bib0270"><span class="elsevierStyleSup">23&#8211;25</span></a> Interestingly&#44; we observed that patients with negative lymph node status &#40;pN0&#41; but positive for circulating tumor cells &#40;CTC&#41; had a shorter OS&#46; These preliminary findings suggest the presence of occult metastatic disease&#46; These results are consistent with previous studies&#44; which have shown that the shedding of tumor cells can occur through pathways independent of lymphatic drainage&#44; indicating that the presence of CTCs in N0 breast cancer predicts disease relapse&#46;<a class="elsevierStyleCrossRef" href="#bib0285"><span class="elsevierStyleSup">26</span></a></p><p id="par0155" class="elsevierStylePara elsevierViewall">In our study&#44; the molecular profiling found an association between <span class="elsevierStyleItalic">RIN3</span> and <span class="elsevierStyleItalic">TP53</span>&#44; which might imply that <span class="elsevierStyleItalic">RIN3</span> variants could induce lung cancer despite the initial normal function of <span class="elsevierStyleItalic">TP53</span>&#46; The role of <span class="elsevierStyleItalic">RIN3</span> regulating tyrosine kinase ABL relates to cytoskeletal remodeling&#44; which affects processes crucial for the spread of lung cancer&#46;<a class="elsevierStyleCrossRef" href="#bib0290"><span class="elsevierStyleSup">27</span></a> A more extensive study integrating COPD and lung cancer patient data may elucidate <span class="elsevierStyleItalic">RIN3&#39;s</span> role in cancer development&#46;</p><p id="par0160" class="elsevierStylePara elsevierViewall">On the other hand&#44; our results suggest that the presence of secondary point mutations affecting <span class="elsevierStyleItalic">TP53</span> together with <span class="elsevierStyleItalic">EGFR</span> mutations is associated with the relapse of the patients&#46; Despite having only two patients with these characteristics&#44; this result aligns with previous works&#44; which have associated concomitant <span class="elsevierStyleItalic">TP53</span> mutations and <span class="elsevierStyleItalic">EGFR</span> mutations with OS and response to treatment&#46; For example&#44; in the study by X&#46; Le et al&#46;&#44; the impact of <span class="elsevierStyleItalic">TP53</span> co-mutations on clinical outcomes in patients with <span class="elsevierStyleItalic">EGFR</span>-mutant advanced NSCLC was examined&#46; This observational study used data from a de-identified database and found that patients with <span class="elsevierStyleItalic">TP53</span> co-mutations had significantly shorter real-world PFS and OS compared to those with wild-type <span class="elsevierStyleItalic">TP53</span> tumors&#46;<a class="elsevierStyleCrossRef" href="#bib0295"><span class="elsevierStyleSup">28</span></a></p><p id="par0165" class="elsevierStylePara elsevierViewall">Furthermore&#44; genetic variants co-occurring in <span class="elsevierStyleItalic">ADAMTSL3</span> and <span class="elsevierStyleItalic">FLT-1</span> &#40;<span class="elsevierStyleItalic">VEGFR-1</span>&#41; suggest involvement in tumor dissemination and prognosis&#44;<a class="elsevierStyleCrossRef" href="#bib0300"><span class="elsevierStyleSup">29</span></a> although the study could not establish their link to CTC presence due to sample size limitations&#46; Investigating <span class="elsevierStyleItalic">ADAMTSL3-FLT1</span> variants in more extensive studies may uncover lung cancer development after COPD&#46;</p><p id="par0170" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">NLRP3</span> and <span class="elsevierStyleItalic">MUC5AC</span>&#44; highly mutated genes in our population&#44; might indicate a potential link to aggressive lung adenocarcinoma and poor outcomes in COPD patients&#44;<a class="elsevierStyleCrossRefs" href="#bib0305"><span class="elsevierStyleSup">30&#44;31</span></a> suggesting their mutational presence could identify COPD patients prone to developing cancer&#46;</p><p id="par0175" class="elsevierStylePara elsevierViewall">Despite the limitations of the small population size&#44; our study observed that baseline tissue variants could predict recurrence better than CTCs&#46; Patients experiencing relapse expressed more potentially impactful &#40;P<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>VUS&#41; variants compared to non-relapsed patients&#46; Less studied genes in the panel contributed to a higher rate of VUS&#44; highlighting the need for further evidence and potential reclassification of these variants&#46;</p></span><span id="sec0080" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0120">Conclusions</span><p id="par0180" class="elsevierStylePara elsevierViewall">This work presents the preliminary results obtained from a small population&#46; These findings aim to generate new hypotheses and lines of research&#46; Our results suggest that combining CTC characterization with the molecular profiling of the tumor might elucidate tumor heterogeneity&#46; A more extensive study with a larger population would enable the identification and stratification of resectable NSCLC patients at higher risk of recurrence&#46;</p></span><span id="sec0085" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0125">Funding</span><p id="par0185" class="elsevierStylePara elsevierViewall">This work was supported by the Health Counseling from the Andalusian <span class="elsevierStyleGrantSponsor" id="gs1">Government and GlaxoSmithKline Spain &#40;GSK Spain&#41;</span> &#40;grant number&#58; <span class="elsevierStyleGrantNumber" refid="gs1">PIP-0192-2020</span>&#41;&#46;</p></span><span id="sec0090" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0130">Authors&#8217; Contributions</span><p id="par0190" class="elsevierStylePara elsevierViewall">M&#46;J&#46;S&#46; conceptualized the study and acquired the funding&#46; M&#46;J&#46;S&#46; and M&#46;dC&#46;G&#46;N&#46; leaded the study&#46; A&#46;G&#46;D&#46;&#44; M&#46;J&#46;S&#46;&#44; M&#46;dC&#46;G&#46;N&#46; designed the experiments&#46; D&#46;dM&#46;P&#46; and A&#46;G&#46;D&#46; processed samples from patients&#46; A&#46;G&#46;D&#46; and M&#46;J&#46;M&#46; carried out the experiments&#46; M&#46;J&#46;M&#46; and C&#46;B&#46;L&#46; recruited the patients involved in the study&#46; J&#46;E&#46;H&#46; supervised clinical management&#46; F&#46;O&#46;&#44; A&#46;G&#46;D&#46; and A&#46;G&#46;M&#46; performed bioinformatic analyses&#46; A&#46;G&#46;D&#46; visualized the data&#46; A&#46;G&#46;D&#46; and M&#46;J&#46;M&#46; wrote the manuscript&#46; J&#46;L&#46;H&#46; assessed the diagnostic of the different histological subtypes of the patients&#46; M&#46;J&#46;S&#46;&#44; M&#46;dC&#46;G&#46;N&#46;&#44; D&#46;dM&#46;P&#46;&#44; D&#46;L&#46;&#44; J&#46;A&#46;L&#46;&#44; C&#46;B&#46;L&#46;&#44; B&#46;A&#46;N&#44; P&#46;J&#46;R&#46; and C&#46;R&#46; conducted manuscript review and editing&#46; All authors have read and agreed to the published version of the manuscript&#46;</p></span><span id="sec0095" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0135">Conflict of Interest</span><p id="par0195" class="elsevierStylePara elsevierViewall">The authors have no competing interests to declare that are relevant to the content of this article&#46;</p></span><span id="sec0100" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0140">Data Availability Statement</span><p id="par0200" class="elsevierStylePara elsevierViewall">Variants found in this work are accessible through ClinVar accession numbers SCV003806285 and SCV003806424&#46;</p></span></span>
      """
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          "titulo" => "Graphical abstract"
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          "identificador" => "xres2271517"
          "titulo" => "Abstract"
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            0 => array:2 [
              "identificador" => "abst0010"
              "titulo" => "Introduction"
            ]
            1 => array:2 [
              "identificador" => "abst0015"
              "titulo" => "Methods"
            ]
            2 => array:2 [
              "identificador" => "abst0020"
              "titulo" => "Results"
            ]
            3 => array:2 [
              "identificador" => "abst0025"
              "titulo" => "Conclusions"
            ]
          ]
        ]
        2 => array:2 [
          "identificador" => "xpalclavsec1893421"
          "titulo" => "Keywords"
        ]
        3 => array:2 [
          "identificador" => "xpalclavsec1893422"
          "titulo" => "Abbreviations"
        ]
        4 => array:2 [
          "identificador" => "sec0005"
          "titulo" => "Introduction"
        ]
        5 => array:3 [
          "identificador" => "sec0010"
          "titulo" => "Material and Methods"
          "secciones" => array:7 [
            0 => array:2 [
              "identificador" => "sec0015"
              "titulo" => "Study Design and Patients"
            ]
            1 => array:2 [
              "identificador" => "sec0020"
              "titulo" => "gDNA extraction from tissue samples and quantification"
            ]
            2 => array:2 [
              "identificador" => "sec0025"
              "titulo" => "Library Preparation"
            ]
            3 => array:2 [
              "identificador" => "sec0030"
              "titulo" => "Sequencing Data Analysis"
            ]
            4 => array:2 [
              "identificador" => "sec0035"
              "titulo" => "Collection of Blood Samples&#46; Enrichment and Isolation of CTCs"
            ]
            5 => array:2 [
              "identificador" => "sec0040"
              "titulo" => "CTC Characterization by Immunofluorescence"
            ]
            6 => array:2 [
              "identificador" => "sec0045"
              "titulo" => "Statistical Analysis"
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          ]
        ]
        6 => array:3 [
          "identificador" => "sec0050"
          "titulo" => "Results"
          "secciones" => array:4 [
            0 => array:2 [
              "identificador" => "sec0055"
              "titulo" => "Detection of Somatic Variants in gDNA From Matched Tumoral&#8211;Peritumoral Tissue Biopsies"
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            1 => array:2 [
              "identificador" => "sec0060"
              "titulo" => "Analyze Mutually Exclusive and Co-occurring Events"
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            2 => array:2 [
              "identificador" => "sec0065"
              "titulo" => "CTCs&#58; Isolation&#44; Characterization&#44; and Prognostic Role"
            ]
            3 => array:2 [
              "identificador" => "sec0070"
              "titulo" => "Integrative Analysis of Tissue Molecular Profiles and Presence of CTCs"
            ]
          ]
        ]
        7 => array:2 [
          "identificador" => "sec0075"
          "titulo" => "Discussion"
        ]
        8 => array:2 [
          "identificador" => "sec0080"
          "titulo" => "Conclusions"
        ]
        9 => array:2 [
          "identificador" => "sec0085"
          "titulo" => "Funding"
        ]
        10 => array:2 [
          "identificador" => "sec0090"
          "titulo" => "Authors&#8217; Contributions"
        ]
        11 => array:2 [
          "identificador" => "sec0095"
          "titulo" => "Conflict of Interest"
        ]
        12 => array:2 [
          "identificador" => "sec0100"
          "titulo" => "Data Availability Statement"
        ]
        13 => array:2 [
          "identificador" => "xack781786"
          "titulo" => "Acknowledgments"
        ]
        14 => array:1 [
          "titulo" => "References"
        ]
      ]
    ]
    "pdfFichero" => "main.pdf"
    "tienePdf" => true
    "fechaRecibido" => "2024-05-07"
    "fechaAceptado" => "2024-08-20"
    "PalabrasClave" => array:1 [
      "en" => array:2 [
        0 => array:4 [
          "clase" => "keyword"
          "titulo" => "Keywords"
          "identificador" => "xpalclavsec1893421"
          "palabras" => array:6 [
            0 => "NSCLC"
            1 => "NGS"
            2 => "CTCs"
            3 => "EZH2"
            4 => "MRD"
            5 => "Liquid biopsy"
          ]
        ]
        1 => array:4 [
          "clase" => "abr"
          "titulo" => "Abbreviations"
          "identificador" => "xpalclavsec1893422"
          "palabras" => array:12 [
            0 => "CK"
            1 => "COPD"
            2 => "CTCs"
            3 => "gDNA"
            4 => "IQR"
            5 => "MRD"
            6 => "NGS"
            7 => "NSCLC"
            8 => "OS"
            9 => "PFS"
            10 => "SEM"
            11 => "VUS"
          ]
        ]
      ]
    ]
    "tieneResumen" => true
    "resumen" => array:1 [
      "en" => array:3 [
        "titulo" => "Abstract"
        "resumen" => "<span id="abst0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0015">Introduction</span><p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">Non-small cell lung cancer &#40;NSCLC&#41; is the most common type of lung neoplasm&#46; Despite surgical resection&#44; it has a high relapse rate&#44; accounting for 30&#8211;55&#37; of all cases&#46; Next-generation sequencing &#40;NGS&#41; based on a customized gene panel and the analysis of circulating tumor cells &#40;CTCs&#41; can help identify heterogeneity&#44; stratify high-risk patients&#44; and guide treatment decisions&#46; In this descriptive study involving a small prospective cohort&#44; we focus on the phenotypic characterization of CTCs&#44; particularly concerning EZH2 expression &#40;a member of the Polycomb Repression Complex 2&#41;&#44; as well as on the mutation profiles of the tissue using a customized gene panel and their association with poor outcomes in NSCLC&#46;</p></span> <span id="abst0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0020">Methods</span><p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">Isolation and characterization of EZH2 on CTCs were evaluated before surgical resection &#40;CTC1&#41; and one month after surgery &#40;CTC2&#41; in resectable NSCLC patients&#46; Targeted NGS was performed using a customized 50-gene panel on tissue samples from a subset of patients&#46;</p></span> <span id="abst0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0025">Results</span><p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">76 patients with resectable NSCLC were recruited&#46; The top mutated genes in the cohort included <span class="elsevierStyleItalic">TP53</span>&#44; <span class="elsevierStyleItalic">FLT1</span>&#44; <span class="elsevierStyleItalic">MUC5AC</span>&#44; <span class="elsevierStyleItalic">EGFR</span>&#44; and <span class="elsevierStyleItalic">NLRP3</span>&#46; Pair of genes that had mutually exclusive mutations was <span class="elsevierStyleItalic">TP53-RIN3</span>&#44; and pairs of genes with co-occurring mutations were <span class="elsevierStyleItalic">CD163-TLR4</span>&#44; <span class="elsevierStyleItalic">FGF10-FOXP2</span>&#44; <span class="elsevierStyleItalic">ADAMTSL3-FLT1</span>&#44; <span class="elsevierStyleItalic">ADAMTSL3-MUC5AC</span> and <span class="elsevierStyleItalic">MUC5AC-NLRP3</span>&#46; CTCs decreased significantly between the two time points CTC1 and CTC2 &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;0001&#41;&#44; and CTCs&#43; patients with high EZH2 expression had an 87&#37; increased risk of death &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;018&#41;&#46;</p></span> <span id="abst0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0030">Conclusions</span><p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">Integrating molecular profiling of tumors and CTC characterization can provide valuable insights into tumor heterogeneity and improve patient stratification for resectable NSCLC&#46;</p></span>"
        "secciones" => array:4 [
          0 => array:2 [
            "identificador" => "abst0010"
            "titulo" => "Introduction"
          ]
          1 => array:2 [
            "identificador" => "abst0015"
            "titulo" => "Methods"
          ]
          2 => array:2 [
            "identificador" => "abst0020"
            "titulo" => "Results"
          ]
          3 => array:2 [
            "identificador" => "abst0025"
            "titulo" => "Conclusions"
          ]
        ]
      ]
    ]
    "NotaPie" => array:1 [
      0 => array:3 [
        "etiqueta" => "1"
        "nota" => "<p class="elsevierStyleNotepara" id="npar0015">Contributed equally&#46;</p>"
        "identificador" => "fn1"
      ]
    ]
    "apendice" => array:1 [
      0 => array:1 [
        "seccion" => array:1 [
          0 => array:4 [
            "apendice" => "<p id="par0215" class="elsevierStylePara elsevierViewall">The followings are the supplementary data to this article&#58;<elsevierMultimedia ident="upi0005"></elsevierMultimedia></p>"
            "etiqueta" => "Appendix A"
            "titulo" => "Supplementary Data"
            "identificador" => "sec0110"
          ]
        ]
      ]
    ]
    "multimedia" => array:9 [
      0 => array:7 [
        "identificador" => "fig0005"
        "etiqueta" => "Fig&#46; 1"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
            "imagen" => "gr1.jpeg"
            "Alto" => 5612
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        ]
        "descripcion" => array:1 [
          "en" => "<p id="spar0030" class="elsevierStyleSimplePara elsevierViewall">Mutational landscape of the sequenced patients&#46; &#40;A&#41; Descriptive flow of all the variants found in the 23 patients &#40;Sankey plot&#41; summarizing all the characteristics&#46; &#40;B&#41; Oncoplot summarizing mutational profile for each patient and gene&#46; Top bar plot represents the number of mutations for each patient filled depending on the consequence of mutation&#46; Right side bar plot illustrates the number of samples affected by mutation in the given gene&#44; filled depending on the consequence of mutation&#46; Clinical stage of each patient is also present below sample names&#44; with corresponding proportions of transitions&#47;transversions&#46;</p>"
        ]
      ]
      1 => array:7 [
        "identificador" => "fig0010"
        "etiqueta" => "Fig&#46; 2"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
            "imagen" => "gr2.jpeg"
            "Alto" => 2580
            "Ancho" => 3733
            "Tamanyo" => 747720
          ]
        ]
        "descripcion" => array:1 [
          "en" => "<p id="spar0035" class="elsevierStyleSimplePara elsevierViewall">Somatic interactions between pairs of genes&#46; Green color is for co-occurrence and pink for mutually exclusive&#44; in boxes&#46; Purple genes are tumor genes&#44; red are inflammation genes&#44; green are COPD-associated genes and black are lung development genes&#46; Number in brackets represents the count of mutations for a given gene&#46; &#42;&#58; <span class="elsevierStyleItalic">p</span>-Value<span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05 for Fisher&#39;s exact test&#46;</p>"
        ]
      ]
      2 => array:7 [
        "identificador" => "fig0015"
        "etiqueta" => "Fig&#46; 3"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
            "imagen" => "gr3.jpeg"
            "Alto" => 935
            "Ancho" => 1500
            "Tamanyo" => 64396
          ]
        ]
        "descripcion" => array:1 [
          "en" => "<p id="spar0040" class="elsevierStyleSimplePara elsevierViewall">Differences in the number of variants between no recurrence vs&#46; recurrence patients&#46; Number of variants presented in both types of patients&#44; grouped by clinical significance&#46; P<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>VUS&#58; Pathogenic variants and VUS&#44; VUS<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>B&#58; VUS and benign variants&#44; P<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>B&#58; Pathogenic variants and benign variants&#44; Total&#58; all types of variants&#46; &#42;&#58; <span class="elsevierStyleItalic">p</span>-value<span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#44; ns is for not significance&#46; Mean with SEM is represented&#46; <span class="elsevierStyleItalic">t</span> test was applied&#46;</p>"
        ]
      ]
      3 => array:7 [
        "identificador" => "fig0020"
        "etiqueta" => "Fig&#46; 4"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
            "imagen" => "gr4.jpeg"
            "Alto" => 1002
            "Ancho" => 1530
            "Tamanyo" => 76697
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        ]
        "descripcion" => array:1 [
          "en" => "<p id="spar0045" class="elsevierStyleSimplePara elsevierViewall">Dynamics of circulating tumor cells &#40;CTCs&#41; and expression of EZH2 in CTCs from NSCLC patients&#46; &#40;A&#41; Differences in the number of CTCs between CTC1 &#40;before surgery&#41; and CTC2 &#40;one month after surgery&#41;&#46; &#42;&#42;&#42;&#42;&#58; <span class="elsevierStyleItalic">p</span> value for Wilcoxon test&#44; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;0001&#46; &#40;B&#41; Comparison of the percentage of expression of EZH2 at CTC1 and CTC2&#46; Abbreviation n&#46;s&#46; is not significant for Mann&#8211;Whitney <span class="elsevierStyleItalic">U</span> test&#44; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;2562&#41;&#46;</p>"
        ]
      ]
      4 => array:7 [
        "identificador" => "fig0025"
        "etiqueta" => "Fig&#46; 5"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
            "imagen" => "gr5.jpeg"
            "Alto" => 3191
            "Ancho" => 3417
            "Tamanyo" => 391667
          ]
        ]
        "descripcion" => array:1 [
          "en" => "<p id="spar0050" class="elsevierStyleSimplePara elsevierViewall">Prognostic role of CTCs and EZH2 expression on CTCs&#46; Kaplan&#8211;Meier at CTC1 of the effect of the number of CTCs based on a cut-off &#40;high<span class="elsevierStyleHsp" style=""></span>&#8805;<span class="elsevierStyleHsp" style=""></span>2&#47;low<span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>2&#41; in the relapse-free survival &#40;<span class="elsevierStyleBold">A</span>&#41; and overall survival &#40;B&#41;&#46; Kaplan&#8211;Meier of the influence of the EZH2 expression cut-off in the progression-free survival &#40;C&#41; and overall survival &#40;D&#41;&#46; Log-Rank &#40;Mantel&#8211;Cox&#41; test was applied in all the panels&#46;</p>"
        ]
      ]
      5 => array:8 [
        "identificador" => "tbl0005"
        "etiqueta" => "Table 1"
        "tipo" => "MULTIMEDIATABLA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "detalles" => array:1 [
          0 => array:3 [
            "identificador" => "at1"
            "detalle" => "Table "
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          ]
        ]
        "tabla" => array:2 [
          "leyenda" => "<p id="spar0060" class="elsevierStyleSimplePara elsevierViewall">The annotation of these variants and the clinical information of the patients that contained them are presented&#46; HGVS&#58; Human Genome Variation Society nomenclature&#44; ID&#58; identifier for the variant&#44; AF&#58; allele frequency&#44; ACMG&#58; The American College of Medical Genetics and Genomics classification&#44; COPD&#58; Chronic Obstructive Pulmonary Disease&#44; P&#58; pathogenic&#44; LP&#58; likely pathogenic&#44; VUS&#58; variants of uncertain significance&#44; F&#58; female&#44; M&#58; male&#44; ADC&#58; adenocarcinoma&#44; SCC&#58; squamous-cell carcinoma&#46;</p>"
          "tablatextoimagen" => array:1 [
            0 => array:2 [
              "tabla" => array:1 [
                0 => """
                  <table border="0" frame="\n
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                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Gene&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Transcript&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">ID&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">AF&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Clinical Significance&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">ACMG Criteria&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Sample&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Gender&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Histology&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Lymph Node Status&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Stage&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">COPD&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Recurrence&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Vital Status&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Cause of Death&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">TP53</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;000546&#46;6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;527G&#62;A&#40;p&#46;Cys176Tyr&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">rs786202962&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">0&#46;192&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">P&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PM1&#44; PM2&#44; PM5&#44; PP2&#44; PP3&#44; PP5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">PCT68&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">F&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">SCC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">N0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">I&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Yes&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Alive&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">TP53</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;000546&#46;6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;584T&#62;C&#40;p&#46;Ile195Thr&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">rs760043106&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;15&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">P&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PM1&#44; PM2&#44; PM5&#44; PP3&#44; PP5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PCT80&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">F&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">ADC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">N0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">I&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Yes&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Alive&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">TP53</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;000546&#46;6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;469G&#62;T&#40;p&#46;Val157Phe&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">rs121912654&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;465&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">VUS&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PM1&#44; PM2&#44; PP3&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PCT33&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">SCC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">N2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">III&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Yes&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Deceased&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Other&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">RB1</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;000321&#46;3&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;1172C&#62;G&#40;p&#46;Ser391&#42;&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">COSV57303024&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;374&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">LP&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PVS1&#44; PM2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="" valign="\n
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                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="" valign="\n
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                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="" valign="\n
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                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">FAM13A</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;014883&#46;4&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;2465C&#62;T&#40;p&#46;Pro822Leu&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">rs377446507&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;174&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">VUS&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PP3&#44; PM2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PCT65&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">SCC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">N1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">II&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Alive&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">RB1</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;000321&#46;3&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;380&#43;2T&#62;C&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">COSV57310513&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;006789&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">LP&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PVS1&#44; PM2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PCT76&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">ADC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">N1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">II&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Alive&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">TP53</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;000546&#46;6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;314G&#62;T&#40;p&#46;Gly105Val&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">rs587781504&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;079&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">LP&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PM1&#44; PM2&#44; PM5&#44; PP2&#44; PP3&#44; PP5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">TP53</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;000546&#46;6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;818G&#62;A&#40;p&#46;Arg273His&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">rs28934576&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;009466&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">P&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PS3&#44; PS4&#44; PM1&#44; PM2&#44; PM5&#44; PP3&#44; PP5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PCT45&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">ADC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">N0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">III&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Alive&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">EGFR</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;005228&#46;5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;2235&#95;2249del&#40;p&#46;Glu746&#95;Ala750del&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">rs121913421&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;081&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">P&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PS1&#44; PM4&#44; PM1&#44; PM2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PCT46&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">F&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">ADC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">N1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">III&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Deceased&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Other&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">TP53</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;000546&#46;6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;622G&#62;T&#40;p&#46;Asp208Tyr&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">COSV52919029&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;031&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">LP&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PM1&#44; PM2&#44; PP3&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PCT47&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">ADC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">N1<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>N2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">III&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Yes&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">No&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Alive&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">EGFR</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;005228&#46;5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;2573T&#62;G&#40;p&#46;Leu858Arg&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">rs121434568&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">P&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">ADC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">N1<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>N2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">III&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Yes&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Yes&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Deceased&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Tumor progression&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">TP53</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NM&#95;000546&#46;6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">c&#46;796G&#62;T&#40;p&#46;Gly266&#42;&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">rs1057519990&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">PVS1&#44; PM1&#44; PM2&#44; PP5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
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                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="" valign="\n
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                  \t\t\t\t">NM&#95;001378902&#46;1&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">c&#46;3388G&#62;A &#40;p&#46;Gly1130Arg&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">PM2&#44; PP2&#44; PP3&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">F&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">ADC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">Yes&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">PVS1&#44; PM2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="" valign="\n
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                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
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                  """
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                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black"><span class="elsevierStyleItalic">p</span>-Value&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Sig&#46;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td-with-role" title="\n
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                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t"><span class="elsevierStyleItalic">Gender</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">0&#46;532&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">n&#46;s&#46;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Male&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">29 &#40;63&#46;0&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">17 &#40;37&#46;0&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Female&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">21 &#40;70&#46;0&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">9 &#40;30&#46;0&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleVsp" style="height:0.5px"></span></td></tr><tr title="table-row"><td class="td-with-role" title="\n
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                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t"><span class="elsevierStyleItalic">Age</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleBold">0&#46;003</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">&#40;<a class="elsevierStyleCrossRef" href="#tblfn0010">&#42;&#42;</a>&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Mean &#40;SD&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">64&#46;80 &#40;7&#46;936&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">70&#46;50 &#40;7&#46;453&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleVsp" style="height:0.5px"></span></td></tr><tr title="table-row"><td class="td-with-role" title="\n
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                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t"><span class="elsevierStyleItalic">Tobacco</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">1&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">n&#46;s&#46;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
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                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Non-smoker&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">9 &#40;64&#46;3&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">5 &#40;35&#46;7&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Smoker&#47;former smoker&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">41 &#40;66&#46;1&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">21 &#40;33&#46;9&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
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                  \t\t\t\t  " colspan="5" align="left" valign="\n
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                  \t\t\t\t"><span class="elsevierStyleVsp" style="height:0.5px"></span></td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">PET &#40;SUVmax&#41;</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">0&#46;996&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">n&#46;s&#46;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
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                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Median &#40;<span class="elsevierStyleItalic">Q</span>1&#44; <span class="elsevierStyleItalic">Q</span>3&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t">7&#46;69 &#40;2&#46;72&#44; 11&#46;78&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">6&#46;04 &#40;3&#46;67&#44; 10&#46;71&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Range&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleVsp" style="height:0.5px"></span></td></tr><tr title="table-row"><td class="td-with-role" title="\n
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                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t"><span class="elsevierStyleItalic">Resection</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">0&#46;796&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">n&#46;s&#46;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
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                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Sublobar resection&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">3 &#40;60&#46;0&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">2 &#40;40&#46;0&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Chemo<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>radiotherapy&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">18 &#40;33&#46;3&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">8 &#40;36&#46;4&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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Article information
ISSN: 03002896
Original language: English
DOI:10.1016/j.arbres.2024.08.006
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