Research Project: Diet and Cardiovascular Health

Location: Jean Mayer Human Nutrition Research Center On Aging

2023 Annual Report

Objectives
Objective 1: Determine the effect of food components and their metabolites, dietary patterns, and lipid-modifying therapies on cardiometabolic risk factors and lipoprotein, sterol, bile acid, and fatty acid metabolism in humans, and using animal, and in vitro models. Sub-objective 1.A: Elucidate the relationship between dietary patterns, with and without statin therapy, on atherosclerotic lesion development and concomitant tissue-specific inflammation using the Ossabaw pig as an experimental model. Sub-objective 1.B: Compare the effects of an isocaloric exchange of simple-carbohydrate (carb), refined-carb, and unrefined-carb on (i) plasma cardiovascular risk factors, (ii) targeted metabolomic and lipidomic markers, and (iii) gut microbiome signatures in humans. Objective 2: Identify novel biomarkers of food intake (e.g., metabolomic, lipidomic, proteomic, and microbiome) and relate them to cardiovascular health. Sub-objective 2.A: Determine the differential effects of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) supplementation, relative to placebo, on (i) plasma measures of cardiovascular risk, and (ii) biomarkers of inflammation and inflammatory cell gene expression in subjects with elevated inflammatory status. Sub-objective 2.B: Evaluate the effects of very long chain omega-3 ('-3) fatty acid supplementation (1.86 g EPA and 1.5 g DHA daily) on the composition and functionality of high density lipoprotein (HDL) subpopulations, and the influence thereof on coronary artery atherosclerotic plaque burden in individuals with stable coronary artery disease on statins.

Approach
Cardiovascular disease continues to be the leading cause of death and disability in the United States. The risk of developing cardiovascular disease increases with age. Preventive measures, especially dietary modification, are more efficacious and cost effective than treatment. However, some dietary recommendations, particularly related to carbohydrate and fat type, are enmeshed in controversy. This controversy undermines public confidence in dietary guidance, thereby impeding efforts to improve the overall quality of the American diet. To address this conundrum, in the next 5 years, the Cardiovascular Nutrition Laboratory will determine the effect of food components and their metabolites, dietary patterns, and lipid-modifying therapies on cardiometabolic risk factors and lipoprotein, sterol, bile acid, and fatty acid metabolism in humans, and using animal and in vitro models. We will accomplish this by determining the effect of dietary modification on cardiovascular health by elucidating the relationships among diet, tissue specific inflammation and atherosclerosis progression using the Ossabaw pig model; investigating the effect of carbohydrate type on cardiovascular disease risk factors and the gut microbiome by conducting human intervention trials; and assessing the relationship among very long chain omega-3 fatty acids (eicosapentaenoic acid and docosahexaenoic acid), inflammation and coronary artery atherosclerotic plaque progression using in vitro and in vivo approaches. More specifically, we will elucidate the relationship between dietary patterns, with and without statin therapy, on atherosclerotic lesion development and concomitant tissue-specific inflammation in the Ossabaw pig, and compare the effects of an isocaloric exchange of simple-carbohydrate, refined-carbohydrate, and unrefined-carbohydrate on cardiovascular risk factors, targeted metabolomic and lipidomic markers, and gut microbiome signatures in humans. We will assess potential complementary and/or synergistic effects between dietary modification and pharmacotherapy intended to reduce cardiovascular disease risk. Additionally, the Cardiovascular Nutrition Laboratory will identify novel biomarkers of food intake (e.g., metabolomic, lipidomic, proteomic, and microbiome) and relate them to cardiovascular health by determining the differential effects of very long chain omega-3 acid supplementation on plasma measures of cardiovascular risk and biomarkers of inflammation and inflammatory cell gene expression in individuals with elevated inflammatory status, and evaluating the effects of very long chain omega-3 fatty acid supplementation on the composition and functionality of high density lipoprotein subpopulations, and the influence thereof on coronary artery atherosclerotic plaque burden in individuals with stable coronary artery disease treated with statins. We will use these data to better understand the relationship between diet and cardiovascular health. The results of the proposed work will help facilitate updating and refining the Dietary Guidelines for Americans intended to support healthy aging.

Progress Report
a) Under Objective 1, in collaboration with ARS scientists at USDA Beltsville, Maryland, we have completed the transcriptomics of brain blood vessels and metabolomics of the temporal region of the brain in Ossabaw pigs randomized to 6 months dietary intervention with either a Heart-healthy diet or Western diet, with or without statins. We found that the brain blood vessels of pigs on Western diet had higher expression of genes related to inflammation, angiogenesis, and apoptosis, while those of pigs on the Heart-healthy diet had higher expression of genes related to cell energetics, neurotransmission, and inflammation resolution pathways. These results suggest a likely contribution of diet to brain pathologies characterized by neuroinflammation and neurodegeneration. b) Under Objective 1, using dietary intake and genetic data from the Framingham Heart Study we empirically generated three dietary patterns (Prudent, Western, and Alcohol) and derived polygenic taste scores using single nucleotide polymorphisms (SNPs) identified from prior genome-wide association studies (GWAS) for taste perception. On average, after adjusting for age, sex, and energy intake, adherence to the Prudent dietary pattern was lower among participants with higher umami polygenic taste scores than the other two dietary patterns. Subsequent analysis at the SNP-level identified one umami-related SNP independently associated with lower Prudent dietary pattern adherence. No significant associations were identified for bitter-related genes and dietary patterns. These data suggest that among community dwelling adults, higher genetic predisposition to perceive umami was associated with lower adherence to a Prudent dietary pattern, suggesting the potential benefit of leveraging knowledge of taste-related genes in precision nutrition. c) Under Objective 1, using 24-hour diet recalls, three diet quality indices were calculated (Alternative Mediterranean Diet, Healthy Eating Index 2015 and Dietary Approaches to Stop Hypertension [DASH]) and associations with cardiometabolic risk factors and visceral adiposity determined. Participants with higher adherence to any of the three heart-healthy dietary patterns had significantly lower BMI and waist circumference compared to lower adherence. Lower glucose and LDL-cholesterol levels, and hepatic fat content, were associated with higher adherence to the DASH diet (higher fruit and lower sugar sweetened beverage intake). No significant associations were observed between cardiometabolic risk factors and adherence to the Alternative Mediterranean Diet and Healthy Eating Index 2015 dietary patterns. d) Under Objective 2, a randomized, crossover study assessing the effects of high-dose (3 g/d) pure eicosapentaenoic acid (EPA) versus docosahexaenoic acid (DHA) on plasma lipids and lipoproteins in older men and postmenopausal women with characteristics of metabolic syndrome and chronic inflammation was conducted. Both EPA and DHA lowered plasma triglyceride concentrations via the activation of lipoprotein lipase. DHA, but not EPA, increased plasma low-density lipoprotein cholesterol concentrations. In addition, EPA and DHA worked differently in men and women. Transcriptomic analysis of monocytes indicated sex specific differences in the expression of several genes. A greater number of genes involved in the interferon pathway and other inflammatory pathways were activated in monocytes isolated from women than men. Our results suggest that the difference in immune functions between women and men may be due to higher activation of interferon in female than male monocytes. e) Under Objective 2, using samples generated from a randomized, crossover study we assessed the effects of diets enriched in stearate (18:0), palmitate (16:0) and oleate (18:1) on gut microbiome composition in post-menopausal women. We found no significant differences in alpha and beta diversity among the three diets. However, the variability in microbial relative abundance was greater after participants consumed the 18:0 and 18:1 compared to the 16:0 diet. Mean relative abundance of Bacterioides dorei, Bacteroides vulgatus, and Ruminococcus were highest after participants consumed the 18:0 relative to the 16:0 diet, and intermediate after the 18:1 diet. The relative abundance of Coprococcus, Parabacteroides distasonis, Coprobacillus and Blautia were higher after participants consumed the 18:1 relative to the 16:0 diet. These microbial species have been associated with several anti-inflammatory pathways and lower bacterial lipopolysaccharide production. f) Under Objective 2, compared were the plasma metabolite profile and lipid-related pathways in samples generated from a randomized, crossover study comparing the effects of diets enriched in partially hydrogenated soybean oil (trans fatty acids) and unmodified oil. Among the known metabolites higher in plasma after the partially hydrogenated soybean oil diet than the soybean oil diet, the majority were phospholipids and diglycerides and triglycerides. Pathway analysis indicated upregulation of phosphatidylcholine synthesis from diglycerides and phosphatidylethanolamine. We identified seven triglyceride related metabolites (TG_56:9, TG_54:8, TG_54:7, TG_54:6, TG_48:5, DG_36:5 and benproperine) as potential biomarkers for partially hydrogenated soybean oil intake. These data indicate that triglyceride-related metabolites were the most affected lipid species, and glycerophospholipid biosynthesis was the most active pathway in response to partially hydrogenated soybean oil compared to soybean oil intake. g) Under Objective 2, compared were the plasma metabolite profiles and their associations with cardiometabolic risk factors in samples generated from a randomized, crossover study comparing the effects of diets enriched in animal and soy proteins. Among the identified metabolites, 58 differed significantly between the animal protein and soy protein diets; the majority were phospholipids (n=36), then amino acids (n=10), xenobiotics (n=7), vitamin/vitamin-related (n=3) and lipids (n=2). Of the top 10 metabolites, amino acid-derived metabolites, phospholipids and xenobiotics comprised the main categories differing due to dietary protein type. Receiver operating characteristic curves confirmed that the top 10 metabolites were potential discriminating biomarkers for animal protein and soy protein-rich diets. These data indicate that amino acid-derived metabolites, phosphatidylethanolamine-derived metabolites and isoflavones were identified as potential metabolite biomarkers distinguishing between dietary protein type.

Accomplishments
1. Cardiovascular disease is the leading cause of death in the United States. Some, but not all very long chain omega-3 fatty acid supplementation studies have shown a reduction in cardiovascular disease risk. There are two major very long chain omega-3 fatty acids, eicosapentaenoic acid (EHA) and docosahexaenoic acid (DHA). Unknown is if they work differently in the prevention of cardiovascular disease and if there are sex specific differences in response to supplementation. ARS-funded researchers in Boston, Massachusetts, conducted a study to compare the effects of EHA and DHA supplementation on a type of immune cell that is actively involved in cardiovascular disease. Researchers found that both EHA and DHA reduced activation of these immune cells. However, compared to DHA, EHA supplementation reduced inflammation pathways only in men. Other responses were only noted in women. These findings suggest sex-dependent effects of EHA and DHA that may be contributing to the conflicting results on cardiovascular disease risk from previous clinical trials.

Review Publications
Zertuche, J., Rabasa, G., Lichtenstein, A.H., Matthan, N., Nevitt, M., Torner, J., Lewis, C.E., Misra, D., Felson, D.T. 2022. Alkylresorcinol, a biomarker for whole grain intake, is not associated with Osteoarthritis: The MOST Study. Osteoarthritis and Cartilage. 30(10):1337-1343. https://doi.org/10.1016/j.joca.2022.07.004.
So, J., Asztalos, B.F., Horvath, K., Lamon-Fava, S. 2022. Ethyl EPA and ethyl DHA cause similar and differential changes in plasma lipid concentrations and lipid metabolism in subjects with low-grade chronic inflammation. Journal of Clinical Lipidology. https://doi.org/10.1016/j.jacl.2022.10.002.
Du, S., Chen, J., Kim, H., Walker, M.E., Lichtenstein, A.H., Chatterjee, N., Ganz, P., Yu, B., Ramachandran, V., Coresh, J., Rebholz, C.M. 2022. Plasma protein biomarkers of healthy dietary patterns: results from the Atherosclerosis Risk in Communities (ARIC) study and the Framingham Heart Study. Journal of Nutrition. https://doi.org/10.1016/j.tjnut.2022.11.008.
Lamon-Fava, S., Liu, M., Dunlop, B.W., Kinkead, B., Schettler, P.J., Felger, J.C., Ziegler, T.R., Fava, M., Mischoulon, D., Rapaport, M.H. 2023. Clinical response to EPA supplementation in patients with major depressive disorder is associated with higher plasma concentrations of pro-resolving lipid mediators. Neuropsychophamacology. https://doi.org/10.1038/s41386-022-01527-7.
Dong, K.R., Beckwith, C.G., Grossman, A., Weiner, D.E., Lichtenstein, A.H. 2022. Utilizing the probation office as an opportunity to screen for cardiometabolic outcomes. Journal of Correctional Health Care. https://doi.org/10.1089/jchc.20.11.0102.
So, J., Tai, A.K., Lichtenstein, A.H., Wu, D., Lamon-Fava, S. 2021. Sexual dimorphism of monocyte transcriptome in individuals with chronic low-grade inflammation. Biology of Sex Differences. 12:43. https://doi.org/10.1186/s13293-021-00387-y.
Zhang, X., Wu, Y., Na, M., Lichtenstein, A.H., Xing, A., Chen, S., Wu, S., Gao, X. 2020. Habitual night eating was positively associated with progress of arterial stiffness in Chinese adults. Journal of the American Heart Association. https://doi.org/10.1161/JAHA.120.016455.
Meng, H., Matthan, N., Angellotti, E., Pittas, A.G., Lichtenstein, A.H. 2020. Exploring the effect of vitamin D3 supplementation on surrogate biomarkers of cholesterol absorption and endogenous synthesis in patients with type 2 diabetes-randomized controlled trial. The American Journal of Clinical Nutrition. https://doi.org/10.1093/ajcn/nqaa149.
Huang, N.K., Buzkova, P., Matthan, N., Djousse, L., Hirsch, C.H., Kizer, J.R., Longstreth, W.T., Mukamal, K., Lichtenstein, A.H. 2021. Associations of serum nonesterified fatty acids with coronary heart disease mortality and nonfatal myocardial infarction: the CHS (cardiovascular health study) cohort. Journal of the American Heart Association. https://doi.org/10.1161/JAHA.120.019135.
Felson, D.T., Misra, D., Lavalley, M., Clancy, M., Chen, X., Lichtenstein, A.H., Matthan, N., Torner, J., Lewis, C.E., Nevitt, M.C. 2021. Fatty acids and osteoarthritis: the MOST study. Osteoarthritis and Cartilage. https://doi.org/10.1016/j.joca.2021.03.006.
Gervis, J., Chui, K.K., Ma, J., Coltell, O., Fernandez-Carrion, R., Sorli, J.V., Barragan, R., Fito, M., Gon Zalez, J.I., Corella, D., Lichtenstein, A.H. 2021. Data-driven clustering approach to derive taste perception profiles from sweet, salt, sour, bitter, and umami perception scores: an illustration among older adults with metabolic syndrome. Journal of Nutrition. https://doi.org/10.1093/jn/nxab160.
Cahoon, D., Shertukde, S., Nirmala, N., Lau, J., Lichtenstein, A.H. 2022. Appraisal of the evidence-base to update dietary reference intake values - lessons from the past, thoughts for the future. Advances in Nutrition. https://doi.org/10.1093/advances/nmac041.
Rodriguez-Morato, J., Matthan, N. 2020. Nutrition and gastrointestinal microbiota, microbial derived secondary bile acids, and cardiovascular disease. Current Atherosclerosis Reports. https://doi.org/10.1007/s11883-020-00863-7.
Wu, Z., Huang, Z., Lichtenstein, A.H., Chen, S., Jin, Y., Na, M., Bao, L., Wu, S., Gao, X. 2021. The risk of ischemic stroke and hemorrhagic stroke in Chinese adults with low-density lipoprotein cholesterol concentrations < 70 mg/dL. BioMed Central (BMC) Medicine. 19:142. https://doi.org/10.1186/s12916-021-02014-4.
Lichtenstein, A.H., Kris-Etherton, P.M., Petersen, K.S., Matthan, N., Barnes, S., Vitolins, M.Z., Li, Z., Sabate, J., Rajaram, S., Chowdhury, S., Davis, K.M., Galluccio, J.M., Gilhooly, C., Legro, R.S., Li, J., Lovato, L., Perdue, L.H., Petty, G., Rasmussen, A.M., Segovia-Siapco, G., Sirirat, R., Sun, A., Reboussin, D.M. 2022. Effect of incorporating 1 avocado per day versus habitual diet on visceral adiposity: A randomized trial. Journal of the American Heart Association. 11(14):e025657. https://doi.org/10.1161/JAHA.122.025657.
Wu, Z., Huang, Z., Lichtenstein, A.H., Jin, C., Chen, S., Wu, S., Gao, X. 2021. Different associations between HDL cholesterol and cardiovascular diseases in people with diabetes mellitus and people without diabetes mellitus: a prospective community-based study. The American Journal of Clinical Nutrition. https://doi.org/10.1093/ajcn/nqab163.
Vadiveloo, M.K., Thorndike, A.N., Lichtenstein, A.H. 2023. Integrating diet screening into routine clinical care - the time is now. Journal of the American Heart Association. https://doi.org/10.1161/JAHA.122.028583.
Djousse, L., Biggs, M.L., Matthan, N., Ix, J.H., Fitzpatrick, A.L., King, I.B., Lemaitre, R.N., Mcknight, B., Kizer, J.R., Lichtenstein, A.H., Mukamal, K.J., Siscovick, D.S. 2021. Serum individual nonesterified fatty acids and risk of heart failure in older adults. Cardiology. 146(3):351-358. https://doi.org/10.1159/000513917.
Ahiawodzi, P., Buzkova, P., Lichtenstein, A.H., Matthan, N., Ix, J.H., Kizer, J.R., Tracy, R.P., Arnold, A., Newman, A.B., Siscovick, D., Djousse, L., Mukamal, K. 2022. The associations of individual and subclasses of non-esterified fatty acids with disability, and mobility limitation in older adults: the Cardiovascular Health Study. Journals of Gerontology. https://doi.org/10.1093/gerona/glac206.
Huang, N.K., Lichtenstein, A.H., Matuszek, G.H., Matthan, N. 2023. Comparison of plasma metabolome response to diets enriched in soybean and partially-hydrogenated soybean oil in moderately hypercholesterolemic adults - a pilot study. Metabolites. https://doi.org/10.3390/metabo13040474.
Faits, T., Walker, M.E., Rodriguez-Morato, J., Meng, H., Gervis, J., Galluccio, J.M., Lichtenstein, A.H., Johnson, W.E., Matthan, N. 2020. Exploring changes in the human gut microbiota and microbial-derived metabolites in response to diets enriched in simple, refined, or unrefined carbohydrate-containing foods: a post hoc analysis of a randomized clinical trial. The American Journal of Clinical Nutrition. https://doi.org/10.1093/ajcn/nqaa254.
Rodriguez-Morato, J., Galluccio, J.M., Dolnikowski, G.G., Lichtenstein, A.H., Matthan, N. 2020. Comparison of the postprandial metabolic fate of U-13C stearic acid and U-13C oleic acid in postmenopausal women. Arteriosclerosis Thrombosis and Vascular Biology. https://doi.org/10.1161/ATVBAHA.120.315260.
Kim, H., Appel, L.J., Lichtenstein, A.H., Wong, K.E., Chatterjee, N., Rhee, E.P., Rebholz, C.M. 2023. Metabolomic profiles associated with blood pressure reduction in response to the DASH and DASH-Sodium dietary interventions. Hypertension. https://doi.org/10.1161/HYPERTENSIONAHA.123.20901.
Kim, H., Lichtenstein, A.H., Ganz, P., Miller, E.R., Coresh, J., Appel, L.J., Rebholz, C.M. 2023. Associations of circulating proteins with lipoprotein profiles: proteomic analyses from the omniheart randomized trial and the atherosclerosis risk in communities (ARIC) study. Clinical Proteomics. https://doi.org/10.1186/s12014-023-09416-x.
Kim, H., Lichtenstein, A.H., Ganz, P., Du, S., Tang, O., Yu, B., Chatterjee, N., Appel, L.J., Coresh, J., Rebholz, C.M. 2023. Identification of protein biomarkers of the dietary approaches to stop hypertension diet in randomized feeding studies and validation in an observational study. Journal of the American Heart Association. https://doi.org/10.1161/JAHA.122.028821.