Dynamic molecular choreography induced by traffic exposure: A randomized, crossover trial using multi-omics profiling

doi:10.1016/j.jhazmat.2021.127359

Journal of Hazardous Materials

Volume 424, Part A, 15 February 2022, 127359

https://doi.org/10.1016/j.jhazmat.2021.127359 Get rights and content

Highlights

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The randomized, crossover trial design had the advantage of strong causal inference.
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First human study to explore the effects of TRAP exposure on exosome transcriptome.
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First-ever multi-omics study to explore the mechanisms of acute TRAP exposure.

Abstract

The biological mechanism of adverse health outcomes related to exposure to traffic-related air pollution (TRAP) needs elucidation. We conducted a randomized, crossover trial among healthy young students in Shanghai, China. Participants wore earplugs and were randomly assigned to a 4-hour walking treatment either along a traffic-polluted road or through a traffic-free park. We conducted untargeted analyses of plasma exosome transcriptomics, serum mass spectrometry-based proteomics, and serum metabolomics to evaluate changes in genome-wide transcription, protein, and metabolite profiles in 35 randomly selected participants. Mean personal exposure levels of ultrafine particles, black carbon, nitrogen dioxide, and carbon monoxide in the road were 2–3 times higher than that in the park. We observed 3449 exosome mRNAs, 58 serum proteins, and 128 serum metabolites that were significantly associated with TRAP. The multi-omics analysis showed dozens of regulatory pathways altered in response to TRAP, such as inflammation, oxidative stress, coagulation, endothelin-1 signaling, and renin-angiotensin signaling. We found that several novel pathways activated in response to TRAP exposure: growth hormone signaling, adrenomedullin signaling, and arachidonic acid metabolism. Our study served as a demonstration and proof of concept on the evidence that associated TRAP exposure with global molecular changes based on the multi-omics level.

Graphical Abstract

Section snippets

Teaser

Our multi-omics analysis provided a preliminary basis for the changes of global circulating molecules in response to traffic-related air pollution.

Study design and participants

We conducted this randomized, crossover trial from October to December 2019. A total of 69 healthy and nonsmoking college students with no history of cardiovascular, respiratory, or allergic diseases were initially recruited from Fudan University of Shanghai, China. We obtained demographic information such as age, sex, height, weight, and disease history at enrollment. All eligible participants were required to complete a two-stage, crossover TRAP exposure treatment: in the first stage, each

Exposure measurement

Real-time personal exposure to traffic-related air pollutants, including UFP, PM_2.5, BC, NO₂, and CO were monitored during each exposure session. The number concentrations of UFP were measured using a unipolar diffusion charger (Philips Aeronsence Nanotracer, Netherlands). PM_2.5 mass concentrations were measured using MicroPEM Personal Exposure Monitors (RTI International, USA). BC was measured using the optical absorption method (MicroAeth Model AE51, AEthlabs, CA). Gaseous pollutants,

Blood collection and sample preparation

We obtained demographic information such as age, sex, height, weight, and past disease history at enrollment. We collected intravenous blood from each participant at the end of each exposure session within one hour. To prevent sample deterioration, those handling samples immediately placed the specimens on ice after collection. We randomly selected 35 participants for the multi-omics analysis due to the limited budget. Multi-level molecular profiling was performed in the randomly selected blood

Isolation and identification of exosome

Exosomes from plasma (2 ml) were isolated using the exoEasy Maxi Kit (Qiagen, Hilden, Germany) according to the manufacturer’s instruction. Exosomes were further identified using transmission electron microscopy (TEM) and Zetaview nanoparticle tracking analysis (NTA). 1) For TEM, 20 μl of the exosome sample was fixed to an electron microscopy mesh grid with 2% glutaraldehyde at room temperature. Then, 2% aqueous solution uranyl acetate was used to stain samples for 5 min, and the filter paper

Sample preparation and data acquisition

Serum samples were thawed on ice, prepared, and analyzed in a randomized order for further detection. Briefly, serum protein concentrations in each sample were quantified using the Micro BCA Protein Assay Kit (Thermo Fisher Scientific, MA). Then, 100 μg protein was extracted from each sample for mass spectrum (MS) analysis. The whole cell lysates of SW620 were used as quality control (QC) samples in proteomics analysis. Then, the QC samples were injected at regular intervals (every 10 samples)

Sample preparation and data acquisition

Serum metabolomic analysis was conducted using both gas chromatography-mass spectrometry (GC-MS) and ultrahigh performance liquid chromatography-mass spectrometry (UPLC-MS) as described in our previous study (HC Li et al., 2017). In brief, GC-MS analysis was conducted using an Agilent gas chromatography incorporated with Agilent 5975C Time-of-Flight mass spectrometer platform (Agilent Technologies, CA). UPLC-MS analysis was conducted with Agilent 1290 infinity UPLC and Agilent Ultra High

Linear regressions to find changes of analytes in response to TRAP

The multi-omic data from transcriptome, proteome, and metabolome were log-transformed before statistical analysis because of their almost log-normal distribution. Analytes (mRNAs, proteins, and metabolites) in less than 1/2 of all detected samples were discarded. Then, values below detection limits were replaced by 1/5 of the minimum value for an analyte using the MetaboAnalyst 5.0 (https://www.metaboanalyst.ca/) (Pang et al., 2021). We conducted a supervised orthogonal partial least

Data description and TRAP exposure

All participants stayed in urban Shanghai and reported no alcohol drinking and medication and dietary supplement use during the study period. Among the 35 randomly selected participants, there are 18 males and 17 females. Baseline demographic characteristics of the 35 participants were comparable to those of the 56 participants (Table S1).

As shown in Fig. 1c, the mean exposure concentrations of UFP, BC, NO₂, and CO for the 35 participants were 2–3 times higher in the Caoxi North Road than in

Discussion

Using the multi-omics approach in this randomized, crossover trial, we detected significant alterations in gene expressions, proteins, and metabolite profiles in response to TRAP. We identified that thousands of analytes were significantly changed in association with short-term exposure to TRAP and individual air pollutants. Our results suggested that UFP, BC, NO₂ and CO were significantly associated with similar numbers of analytes in the proteome, transcriptome, and metabolome, which were

CRediT authorship contribution statement

Xihao Du and Qingli Zhang analyzed the data and wrote the draft. Xihao Du, Yixuan Jiang, Qingli Zhang, Xinlei Zhu, Cong Liu, and Yue Niu collected the data. Huichu Li, Yang Zhang, John Ji, Chao Jiang, Jing Cai, Renjie Chen, and Haidong Kan revised the manuscript. Haidong Kan and Renjie Chen designed and supervised the study.

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgment

This work was supported by the National Natural Science Foundation of China (92043301, 91843302, and 82030103) and the Fifth Round of the Three-Year Public Health Action Plan of Shanghai (GWV-10.1-XK08 and GWV-10.2. XD17).

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Citation Excerpt :
We performed a randomized, crossover trial from October to December 2019 in Shanghai, China. The details of our study design have been reported in our previous research (Du et al., 2022). Briefly, we recruited 69 nonsmoking and healthy college students as study participants.
Traffic-related air pollution (TRAP) has been associated with changes in gene-specific DNA methylation. However, few studies have investigated impact of TRAP exposure on genome-wide DNA methylation in circulating blood of human.
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The average concentrations of traffic-related air pollutants including black carbon, ultrafine particles, carbon dioxide, and nitrogen dioxide were 2–3 times higher in the road compared to those in the park. Methylation levels of 68 CpG loci were significantly changed (false discovery rate < 0.05) following TRAP exposure, among which 49 were hypermethylated and 19 were hypomethylated. The annotated genes based on the differential CpGs loci were related to pathways in cardiovascular signaling, cytokine signaling, immune response, nervous system signaling, and metabolism.
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¹: Xihao Du and Qingli Zhang contributed equally to this work.

View full text

Dynamic molecular choreography induced by traffic exposure: A randomized, crossover trial using multi-omics profiling

Highlights

Abstract

Graphical Abstract

Section snippets

Teaser

Study design and participants

Exposure measurement

Blood collection and sample preparation

Isolation and identification of exosome

Sample preparation and data acquisition

Sample preparation and data acquisition

Linear regressions to find changes of analytes in response to TRAP

Data description and TRAP exposure

Discussion

CRediT authorship contribution statement

Declaration of Competing Interest

Acknowledgment

Cell

Chem. Biol. Interact.

Toxicol. Lett.

Prog. Retin. Eye. Res.

Gene

Environ. Int.

Environ. Int.

Biol. Psychiatry

Lancet

Environ. Int.

J. Hazard. Mater.

Sci. Total. Environ.

Growth hormone’s links to cancer

Endocr. Rev.

Traffic-related air pollution exposure over a 5-year period is associated with increased risk of asthma and poor lung function in middle age

Eur. Respir. J.

The diacetyl-exposed human airway epithelial secretome: new insights into flavoring-induced airways disease

Am. J. Respir. Cell Mol. Biol.

Fine particulate air pollution and the expression of micrornas and circulating cytokines relevant to inflammation, coagulation, and vasoconstriction

Environ. Health Perspect.

Near-roadway air pollution exposure and altered fatty acid oxidation among adolescents and young adults - the interplay with obesity

Environ. Int.

Inhibition of mpo (myeloperoxidase) attenuates endothelial dysfunction in mouse models of vascular inflammation and atherosclerosis

Arterioscler. Thromb. Vasc. Biol.

Global burden of 369 diseases and injuries in 204 countries and territories, 1990-2019: a systematic analysis for the global burden of disease study 2019

Lancet

Cardiovascular risks from fine particulate air pollution

N. Engl. J. Med

Exposure to traffic-related air pollution and serum inflammatory cytokines in children

Environ. Health Perspect.