PILOT: Center for Appalachian Research in Envrionmental Sciences: Interactions Between Diet and Environmental Exposures During Interventions to Reduce Cardiovascular Disease Risk

Individuals living in Appalachia and in particular eastern Kentucky have far higher risk of cardiovascular diseases than the general population. Heritable genetic and behavioral factors (for example poor diet and smoking) are major determinants of cardiovascular disease risk. Substantial evidence also associates involuntary exposure to environmental agents, for example air pollution and environmental chemicals in foods and drinking water with risk of cardiovascular and metabolic diseases. These exposures are of great concern to UK-CARES stakeholders in Appalachian Kentucky and could contribute to the increased risk of disease in these individuals. However, essentially nothing is currently known about what environmental chemicals this relevant population are exposed to or how this might affect their cardiovascular health. This pilot project will leverage substantial existing and ongoing investments of NIH funds in longitudinal/ interventional studies to reduce cardiovascular disease risk in individuals living in Appalachian Kentucky. We will conduct an exploratory study of exposure to relevant environmental chemicals by making measurements in plasma samples from subjects enrolled in the Heart Health intervention (HHI) and Rural Intervention for Caregivers Heart Health (RICH) studies directed by Dr Moser. These are two-group randomized controlled trials with ~600 subjects to test the efficacy of a lifestyle/behavioral intervention on cardiovascular disease risk factors at 4 and 12 months after initiation of the intervention. The RICH study enrolls primary caregivers of adult patients with chronic diseases Dr Morris and staff using instrumentation in the UK-CARES Analytical Core will conduct analytical measurements of environmental chemicals and other relevant biomarkers. A statistician, Dr Charnigo, who has extensive relationships with these investigators will provide support for the project. The primary focus of the pilot phase of this study will be to measure levels of per and poly fluorinated alkyl substances (PFAS) in plasma samples from these cohorts. These are surfactant chemicals found in clothing, furniture and food packaging. They can be detected in the blood of almost all Americans. Cross sectional epidemiological studies associate exposure to PFAS with elevated circulating levels of cholesterol and triglycerides which are established cardiovascular risk factors. We will examine associations between PFAS and plasma lipids in the RICH and HOF cohorts, determine in these associations are altered by the intervention and measure biomarkers of dietary cholesterol absorption and cholesterol synthesis to investigate the mechanism(s) linking PFAS exposures to elevated plasma cholesterol levels. Statement of work. 1. Establish and validate HPLC coupled electrospray ionization mass spectrometry assays for 12 common PFAS with quantitation accomplished using mass labeled surrogates and internal standards (adaptations of EPA method 537). 2. Establish and validate plate based sample processing methods for measurement of PFAS in human plasma 3. Conduct a pilot study using HHI samples to evaluate the technical robustness of these methods 4. Execute measurements of PFAS in ~400 samples from the control and intervention groups of the RICH study 5. Analyze data using multivariate statistics to examine associations between PFAS and other cardiovascular disease risk factors including elevated plasma lipids. Investigate impact of the risk reduction intervention on circulating PFAS levels. 6. Develop and validate GC MS/MS methods for measurement of total cholesterol, plant phytosterols and cholesterol metabolites in human plasma including lathosterol and sistosterol (this method is already in hand) 7. Pilot study using HHI samples to evaluate technical robustness of these methods 8. Execute measurements of sterols in PFAS in ~400 samples from the control and intervention groups of the RICH study 9. Analyze data using multivariate statistics to examine associations between sterols PFAS and other cardiovascular disease risk factors including elevated plasma lipids. Investigate impact of the risk reduction intervention on sterol biomarkers. 10. Collect plasma samples from HHI cohort 11. Use methods discussed above to measure and analyze sterol biomarker and PFAS levels in these samples 12. Preparation of data for publication and use in grant proposals (includes upcoming application for a competitive supplement to RICH study for measurements of biomarkers of diet.