There is a need to adopt a global epidemiological approach to study the environmental and genetic causes of cardiovascular disease (CV) and to evaluate the hypothesis that epigenetic alterations in genes associated with cardiovascular risk act as partial mediators of the risk associated with environmental exposures. The proposed project will enable: 1) to evaluate the role of metals as environmental determinants of subclinical arteriosclerosis (Objective 1); 2) characterise the mediating role of DNA methylation and hydroxymethylation alterations in candidate genes related to arteriosclerosis and functionally validate the identified regions (Objective 2); and (3) identify how genetic variation interacts with alterations in DNA methylation and hydroxymethylation associated with possible vascular toxicity of metals (Objective 3). For this we will use biological samples, survey data, physical examination and subclinical arteriosclerosis determinations (coronary calcium, intimate-medium thickness and presence of plaque in carotids, iliacs and femorals), which were obtained in 2011-2013 in a cohort of workers in a car assembly factory (AWHS study). Cadmium, inorganic arsenic, tungsten and antimony in urine will be measured from 2800 individuals. Only 1000 individuals have total blood to measure lead, mercury and isolate DNA for polymorphism determination and epigenetic markers. The proposed lines of research will not only provide clinically relevant evidence to understand the causes of cardiovascular disease, but could also contribute to the development of future preventive, diagnostic and therapeutic strategies to promote healthy aging.