title: Blood hypomethylation of inflammatory genes mediates the effects of metal-rich airborne pollutants on blood coagulation.
abstract: OBJECTIVE
  Recent investigations have associated airborne particulate matter (PM) with increased coagulation and thrombosis, but underlying biological mechanisms are still incompletely characterised. DNA methylation is an environmentally sensitive mechanism of gene regulation that could potentially contribute to PM-induced hypercoagulability. We aimed to test whether altered methylation mediates environmental effects on coagulation.
  
  
  METHODS
  We investigated 63 steel workers exposed to a wide range of PM levels, as a work-related condition with well-characterised prothrombotic exposure. We measured personal PM10 (PM≤10 µm in aerodynamic diameter), PM1 (≤1 µm) and air metal components. We determined leukocyte DNA methylation of NOS3 (nitric-oxide-synthase-3) and EDN1 (endothelin-1) through bisulfite-pyrosequencing and we measured ETP as a global coagulation-activation test after standardised triggers.
  
  
  RESULTS
  ETP increased in association with PM10 (β=20.0, 95% CI 3.0 to 37.0), PM1 (β=80.8 95% CI 14.9 to 146.7) and zinc (β=51.3, 95% CI 0.01 to 111.1) exposures. NOS3 methylation was negatively associated with PM10 (β=-0.2, 95% CI -0.4 to -0.03), PM1 (β=-0.8, 95% CI -1.4 to -0.1), zinc (β=-0.9, 95% CI -1.4 to -0.3) and iron (β=-0.7, 95% CI -1.4 to -0.01) exposures. Zinc exposure was negatively associated with EDN1 (β=-0.3, 95% CI -0.8 to -0.1) methylation. Lower NOS3 (β=-42.3; p<0.001) and EDN1 (β=-14.5; p=0.05) were associated with higher ETP. Statistical mediation analysis formally confirmed NOS3 and EDN1 hypomethylation as intermediate mechanisms for PM-related coagulation effects.
  
  
  CONCLUSIONS
  Our study showed for the first time, that gene hypomethylation contributes to environmentally induced hypercoagulability.
authors.affiliation: Department of Clinical and Community Sciences, Università degli Studi di Milano and IRCCS Maggiore Hospital, Mangiagalli and Regina Elena Foundation, Milan, Italy.
authors.forename: Letizia
authors.initials: L
authors.lastname: Tarantini
authors.forename: Matteo
authors.initials: M
authors.lastname: Bonzini
authors.forename: Armando
authors.initials: A
authors.lastname: Tripodi
authors.forename: Laura
authors.initials: L
authors.lastname: Angelici
authors.forename: Francesco
authors.initials: F
authors.lastname: Nordio
authors.forename: Laura
authors.initials: L
authors.lastname: Cantone
authors.forename: Pietro
authors.initials: P
authors.lastname: Apostoli
authors.forename: Pier Alberto
authors.initials: PA
authors.lastname: Bertazzi
authors.forename: Andrea A
authors.initials: AA
authors.lastname: Baccarelli
mesh_terms: D000328:Adult; D000393:Air Pollutants / Q000009:adverse effects*; D001777:Blood Coagulation / Q000187:drug effects* / Q000235:genetics; D001778:Blood Coagulation Disorders / Q000209:etiology* / Q000235:genetics / Q000378:metabolism; D016001:Confidence Intervals; D019175:DNA Methylation; D019332:Endothelin-1 / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007221:Industry; D007249:Inflammation / Q000235:genetics / Q000378:metabolism; D019570:Inhalation Exposure; D007962:Leukocytes / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D009784:Occupational Diseases / Q000209:etiology / Q000235:genetics / Q000378:metabolism; D016273:Occupational Exposure / Q000009:adverse effects*; D052638:Particulate Matter / Q000009:adverse effects*; D013927:Thrombosis / Q000453:epidemiology / Q000209:etiology / Q000378:metabolism; D015032:Zinc / Q000009:adverse effects*
chemical_list: D000393:Air Pollutants; D019332:Endothelin-1; D052638:Particulate Matter; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; D015032:Zinc