FOLATE METABOLISM AND REQUIREMENTS: IMPACT OF OBESITY

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 0229551
• Project Start Date: Jul 1, 2012
• Project End Date: Jul 1, 2017
• Program Code: [(N/A)]- (N/A)

Recipient Organization
UNIVERSITY OF GEORGIA
200 D.W. BROOKS DR
ATHENS,GA 30602-5016

Performing Department
College of Family & Consumer Sciences

Non Technical Summary
Folate is a water-soluble vitamin that is required for DNA synthesis. All cells require folate for cell division and it is especially important during periods of rapid growth. Fetal neural tube development is dependent on maternal folate status during the initial weeks of gestation. Periconceptional supplementation with folate has been shown to reduce the risk of neural tube defects (NTDs). Accordingly, current recommendations established in 1998, specify that all women capable of becoming pregnant consume 400 mcg of folate daily from supplements, fortified foods, or both, in addition to consuming food folate from a varied diet. However, despite fortification, the risk for NTDs increases with higher pre-pregnancy weight. In addition, an association between increased body weight and decreased intake of supplemental folic acid has been reported. The percentage of women of childbearing age not taking folic acid supplements increases with increasing BMI. To further compound the problem, even when intakes are comparable to normal weight women, obesity is associated with lower serum folate in women of childbearing age. Over 50% of women of reproductive age in the US are overweight or obese. Only 1 in 4 meet the IOM recommended 400 mcg of folic acid a day. Therefore, the objective of this study is to determine if there are differences in the metabolic response to a single dose of folate between normal weight and obese women of childbearing age. The two groups of women will be given one of two doses of the vitamin, either the currently recommended amount, 400 mcg, or twice that amount. Blood samples will be taken before the vitamin is given and at various points afterwards to determine how fast the vitamin is cleared from the blood. Body composition measurements, and diet and physical activity information will also be taken to determine if the rate of clearance is influenced by these factors. In a second study, normal weight and obese women of reproductive age will be fed a diet low in folate in order to deplete their serum folate concentrations. Subsequently, a folate-adequate diet will replete the folate status of participants. Blood folate and folate-related compounds will be measured and the effect of folate depletion will be compared between normal weight and obese women. Results of this research should help determine if folate requirements are higher for obese than for non-obese women. The long-term goal of this and follow-up studies is to generate data that will contribute to the evidence base used by scientific advisory panels to determine whether dietary folate intake recommendations should be based on body weight.

Animal Health Component

(N/A)

Research Effort Categories

Basic

(N/A)

Applied

(N/A)

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
702	6099	1010	100%

Knowledge Area
702 - Requirements and Function of Nutrients and Other Food Components;

Subject Of Investigation
6099 - People and communities, general/other;

Field Of Science
1010 - Nutrition and metabolism;

Keywords

folate

folic acid

obesity

one-carbon

nutrient requirements

Goals / Objectives
The overarching goal of this research is to provide evidence that obesity increases the folate requirement to maintain normal serum folate, the body pool most likely to impact the developing neural tube during the first weeks of embryonic development. The proposed studies will examine the effects of obesity on folate pharmacokinetics, both short- and long-term, as well as on folate-related biomarkers. The specific objectives of this work are 1) To determine short-term response in serum folate levels after a single dose of folic acid in normal weight and obese women; 2) To evaluate the response to a controlled dietary folate depletion-repletion protocol in normal and obese women; 3) To investigate the associations between the total adipose tissue, as well as body fat distribution, and folate status and clearance from serum; 4) To explore epigenetic biomarkers of folate status. In the first study, we expect to see differences between the non-obese and obese women in short-term metabolic responses to a 400 mcg folic acid dose. We expect the pharmacokinetic differences in folate response to between non-obese and obese women to be attenuated when the dose administered is 800 mcg. In the second study, we expect to observe different responses between normal weight and obese women to the dietary folate depletion-repletion protocol. We expect to show that obese women become depleted to a greater extent and/or at a faster rate than normal weight women. It is also expected that the 400 mcg/d DFE provided in the repletion diet will not bring serum folate levels in the obese group to the same levels as those in the normal weight group and that serum folate at week 14 will be lower in the obese women, compared to the normal weight group. In deepening our investigation of folate-obesity interactions, we intend to determine whether excess adiposity or increased lean mass have a greater influence on folate pharmacokinetic differences between normal weight and obese women of reproductive age. Furthermore, we are interested in whether the site of the adiposity (central or peripheral) influences any observed effect. At present very little is known about folate metabolism in adipose tissue. The metabolic activities of adipose tissue may vary according to location of adiposity; consequently, the role of folate may be different in visceral than subcutaneous adipose. We also expect to gain information on contributions of dietary folate intake to folate status in normal weight and obese women. We expect to observe lower serum folate levels in the obese group despite similarities in folate intake, supporting our hypothesis of a higher folate requirement in obesity. A novel outcome of the proposed research is to identify any effects of obesity on the low folate-associated decrease in methylation. We hope to identify genes that are more prone to hypomethylation under low folate conditions. The methylation status of these genes may serve as biomarkers for early identification of impairment of folate metabolism. Overall, this research may provide evidence for a revision of the current folate requirements, which do not take BMI into consideration.

Project Methods
Response to a single, acute dose of folic acid, as well as to a long-term dietary folate depletion and repletion, will be compared in normal weight and obese women of reproductive age. Anthropometric measurements of height, weight, and waist circumference will be determined for calculation of BMI and as an indication of degree of central adiposity. Height will be measured using a vertical standing stadiometer to the nearest 0.1 cm. Weight will be measured to the nearest 0.1 kg by an electronic scale. These parameters will be measured before the first blood draw, to verify eligibility based on body mass index. Height and weight measurements will be used to calculate the participant's BMI using the Center for Disease Control's website BMI calculator tool. Body fat mass (g), fat-free mass (g), and percent body fat will be measured using the BOD POD body composition tracking system (COSMED, Concord, CA). The BOD POD technology uses air-displacement plethysmography applying whole body densitometry to measure body composition. Participants will be asked to sit still and breath as normal as possible while seated in the testing chamber for the body volume measurements. BOD POD data will be complemented with measurements collected using DXA, which will also provide information on the participants' bone health including total bone area, bone mineral content and bone density. In addition, the DXA will provide data on fat distribution. The participants in our studies will also complete 24-hr dietary recalls for three non-consecutive days including one weekend day, from their home or other location, using a web-based automated self-administered program (the multi-pass Automated Self-administered 24-hour Recall (ASA24) system, hosted through the National Cancer Institute website). The recorded intakes are automatically analyzed using the USDA Food and Nutrient Database for Dietary Studies, which provides an accurate assessment of individual caloric and nutrient intake. The folate concentrations of the blood specimens will be determined using the Lactobacillus casei microbiological assay in a 96-well microplate system. The serum folate concentrations will be used to generate time-concentration curves, which will be used to calculate pharmacokinetic parameters such as clearance rates and volume of distribution. A two-way analysis of variance will be carried out to test the main effects of folate dose and obesity stratum as well as their interaction on the pharmacokinetic measures. The effects of confounding variables (i.e. lean body mass, dietary intake, other health/nutritional indicators, folate-related polymorphisms) will be explored using two-way analysis of covariance. Unpaired t-tests will be used to evaluate differences among treatment groups with respect to folate-related metabolites and masses of folate pools. Pearson's correlations will be employed to examine associations between adiposity and kinetic parameters.

Progress 10/01/16 to 07/01/17

Outputs
Target Audience:National and international scientific community, national and international colleagues in folate field, medical and allied professionals, non-pregnant and pregnant women of reproductive age. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Research activities during this project period provided opportunities for professional development for a masters/dietetic intern student and several undergraduate majors in nutrition science and dietetics. During this period, the students continued their involvement in the folic acid supplementation trial in pregnant women conducted in collaboration with Athens Regional Midwifery Practice. Specifically, they assisted in sample and data analysis related to cell-type associated DNA methylation in normal and obese women of reproductive age, whole blood global DNA methylation response to folic acid supplementation during pregnancy, and response of one carbon biomarkers in maternal and cord blood to folic acid dose during pregnancy. All trainees attended journal club discussions of current literature related to folate status, folic acid supplementation, nutrient requirements during pregnancy and other topics pertinent to the research project. They also assisted in the preparation of publications for submission to peer-reviewed journals and/or prepared abstracts and research posters which they presented at local or state-wide conferences. All students involved in the project presented at professional meetings and/or research symposia. Research data from the project provided the basis for one student thesis. How have the results been disseminated to communities of interest?The results have been disseminated through peer review publications and presentations at local and state-wide scientific conferences such as Experimental Biology (annual meeting of American Society for Nutrition) and the University of Georgia CURO undergraduate research symposium. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Recent data indicates that almost 60% of women of reproductive age are overweight or obese and that obese women are at higher risk for having a neural tube defect (NTD) affected pregnancy. Current recommendations for optimal intake of folate for women of this age group are based on periconceptional levels shown to be effective for NTD risk reduction, but do not take body weight into consideration. The purpose of this project is to evaluate the impact of obesity on folate metabolism and requirements. The project has four objectives, two of which were addressed during this project period: 1) To determine response to folic acid supplementation in normal weight and obese women and 2) To explore epigenetic biomarkers of folate status. Research during this project period focused on completion of and data analysis for the folic acid supplementation dose response study in pregnant women conducted in collaboration with Athens Regional Midwifery Practice. This was a double-blind randomized controlled trial in healthy pregnant women (BMI 18.5 - 29.9 kg/m2, age 18 - 40 yr) receiving prenatal care at Athens Regional Midwifery clinic (ARMC). The primary goal of the study was to determine relative changes in blood folate and DNA methylation levels in response to two different supplemental doses of folic acid one of which provides the Institute of Medicine's recommended folate intake (400 mcg/d folic acid) for pregnant women and the second which provides a higher dose (800 mcg/d folic acid) as routinely taken by pregnant women in over-the-counter prenatal supplements. Pregnant women were recruited at their first prenatal visit (< 12 weeks) and randomly assigned to take supplements containing either 400 or 800 mcg/d folic acid for the remainder of the pregnancy. Blood samples were taken at baseline, 28 and 36 weeks gestation and from the mother and infant (cord blood) at delivery for determination of folate status and DNA methylation. Recruitment for the study began in mid-July 2014 with 49 women initially enrolled in the study and 28 completed the intervention. As the intervention period ended with the final delivery in August 2015, the primary focus of this project period was on analysis of blood samples for folate biomarkers and DNA methylation response and statistical analysis of the resultant data. The first delivery samples were collected in mid-February, and the last in mid-August. Biological samples were stored at -80?C and sent upon completion of sample collection to collaborating investigators at the University of Florida (for RBC and serum folate analysis), Cornell University (global DNA methylation, choline, unmetabolized folic acid), and Emory University (genome-wide and gene-specific DNA methylation analysis). It is anticipated that this investigation will provide new evidence that will link and compare observed changes in DNA methylation of specific genes associated with birth defects and chronic disease in response to these two prenatal folic acid doses and characterize the impact of maternal obesity on the epigenomic response.

Publications

• Type: Journal Articles Status: Published Year Published: 2017 Citation: Park, H. J., Bailey, L. B., Shade, D. C., Hausman, D. B., Hohos, N. M., Meagher, R. B., Kauwell, G. P., Lewis, R. D.,Smith, A. K. Distinctions in gene-specific changes in DNA methylation in response to folic acid supplementation between women of normal weight and obesity. Obes. Res. Clin. Prac. Jul 18. pii: S1871-403X(17)30062-5. doi: 10.1016/j.orcp.2017.06.004
• Type: Journal Articles Status: Awaiting Publication Year Published: 2017 Citation: Shade, D. C., Park, H. J., Hausman, D. B., Hohos, N. M., Meagher, R. B., Kauwell, G. P., Kilaru, V., Lewis, R. D.,Smith, A. K., Bailey, L. B. DNA methylation changes in whole blood and CD16+ neutrophils in response to chronic folic acid supplementation in women of childbearing age.. Int. J. Vitam. Nutr. Res. (In Press)
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Gordon, J. D., Hausman, D. B., Bailey, L. B., Lewis, R. D., Laing, E. M., Park, H. J. Impact of gluten-containing foods on folate intake of women of childbearing age. The Digest, 51: 2-7
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Park, H. J., Knight, A. K., Fleming, J. M., Hausman, D. B., Caudill, M. A., Malysheva, O. V., Kauwell, G. P.A., Sokolow, A., Fisher, S., Smith, A. K., Bailey, L. Association Between One-carbon Metabolism Indices and DNA Methylation Status in Maternal and Cord Blood FASEB J 31:802.28.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Hohos, N., Smith, A. K., Kilaru, V., Park, H. J., Hausman, D., Bailey, L. B., Lewis, R. D., Meagher, R. B., Differential DNA Methylation between Obese and Normal Weight Women in CD4+ and CD8+ T cells, and CD16+ neutrophils FASEB J 30:431.1
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Park, H. J., Hausman, D., Fleming, J., Andersen, V., Rosa, G., Kauwell, G. P. A., Sokolow, A., Caudill, M. A., Malysheva, O., Fisher, S., Bailey, L. B., Folate and Choline Metabolite Changes in Maternal and Cord Blood in Response to Folic Acid Supplementation Dose During Pregnancy FASEB J 30:1172.10
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Fleming, J., Park, H. J., Hausman, D., Rosa, G., Andersen, V., Kauwell, G. P. A., Caudill, M. A., Malysheva, O., Fisher, S., Bailey, L. B., Unmetabolized Folic Acid Response to Folic Acid Supplementation Dose During Pregnancy FASEB J 30:1172.13
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Hausman, D., Holder, A., Park, H. J., Rosa, G., Andersen, V., Fleming, J., Kauwell, G. P. A., Caudill, M. A., Malysheva, O., Fisher, S., Bailey, L. B., Vitamin B12 Status and Infant Outcome in a Community-Based Folic Acid Intervention Study FASEB J 30:1172.7
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: CURO Symposium. UGA Center for Undergraduate Research Opportunities. Neutrophil and granulocyte methylation in normal weight and obese individuals. Jenissa Gordon, Natalie Hohos, Deanna C. Shade, Dorothy B. Hausman, Gail P. A. Kauwell, Hea Jin Park Lynn B. Bailey, Athens, GA, April 4, 2016.
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: CURO Symposium. UGA Center for Undergraduate Research Opportunities. Blood Folate and Whole Blood Global Methylation Response to Folic Acid Supplementation Dose During Pregnancy. Joanna Yang, Dorothy B. Hausman, Jennifer Fleming, Gail P. A. Kauwell, Marie A. Caudill, Hea Jin Park, Lynn B. Bailey. Athens, GA, March 30, 2015.
• Type: Theses/Dissertations Status: Published Year Published: 2016 Citation: Jennifer Fleming. Response of One-Carbon Biomarkers in Maternal and Cord Blood to Folic Acid Dose during Pregnancy. Masters Thesis. University of Georgia

Progress 07/01/12 to 07/01/17

Outputs
Target Audience:National and international scientific community, national and international colleagues in folate field, medical and allied professionals, non-pregnant and pregnant women of reproductive age. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Over the five-year project term, research activities of this project provided opportunities for professional development for a temporary assistant professor, a postdoctoral associate, four masters/dietetic intern students, a medical student and numerous undergraduate majors in biology, nutrition science and dietetics. The assistant professor and postdoctoral associate assisted in the development, implementation and coordination of three studies of the acute or chronic effects of folic acid supplementation in normal weight and obese non-pregnant and pregnant women. They, as well as the students at all levels, gained experience in subject recruitment and retention, collaboration and coordination with clinical staff, compliance monitoring, sample collection and processing, sample preparation for analysis, data entry, statistical analysis and many other skills involved in successful conduct of clinical research studies. All trainees attended journal club discussions of current literature related to folate status, folic acid supplementation, nutrient requirements during pregnancy and other topics pertinent to the research project. They also assisted in the preparation of publications for submission to peer-reviewed journals and/or prepared abstracts and research posters. All presented research findings at scientific or clinical conferences at national, local or state-wide level. How have the results been disseminated to communities of interest?The results have been disseminated through peer review publications and presentations at local and state-wide scientific conferences such as Experimental Biology (annual meeting of American Society for Nutrition), Obesity Week (annual meeting of The Obesity Society, the Medical College of Georgia/University of Georgia Medical Partnership Student Research Symposium, the Georgia Academy of Nutrition and Dietetics Annual Conference and Exhibition, the SEC Symposium: Prevention of Obesity: Overcoming a 21st Century Public Health Challenge. The Academic Initiative of the SEC, the Southern Translational Education and Research, STaR Conference and the University of Georgia CURO undergraduate research symposium. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Recent data indicates that almost 60% of women of reproductive age are overweight or obese and that obese women are at higher risk for having a neural tube defect (NTD) affected pregnancy. Current recommendations for optimal intake of folate for women of this age group are based on levels shown to be effective for maintaining sufficient folate status during the periconceptional period for NTD risk reduction. These recommendations do not take body weight into consideration. The purpose of this project is to evaluate the impact of obesity on folate metabolism and requirements. Three clinical studies addressing project goals were performed during this five-year project. Two project goals, to determine short-term response in serum folate levels to a single dose of folic acid in normal weight and obese women and to investigate associations between adipose tissue/body fat distribution and folate status and clearance from serum, were addressed in a folic acid pharmacokinetics study. Results of the study indicated that the overall serum response to a single 400 ug dose of folic acid was lower in obese as compared to normal-weight women of reproductive age which suggests that body distribution of folate is affected by obesity. In addition, the serum folate response differed according to percent body fat, with a higher post dose peak observed for women with normal (<32%) body fat. Together, these findings indicate that the rate of absorption and distribution of folate differs between normal weight and obese women of child bearing age which is specifically impacted by adiposity and may reduce the amount of folate available to the developing embryo should pregnancy occur. Epigenetic biomarkers of folate status were explored in two clinical trials in women of reproductive age. In the first, nonpregnant women with normal weight and obesity were supplemented with 800 ug/d folic acid for eight weeks. Results indicated a distinctive pattern of DNA methylation response to the supplementation in normal weight and obese women, including differences in changes in DNA methylation in genes related to NTD risk and folate metabolism between the two groups. The second study investigated the impact of one-carbon metabolism indices during pregnancy on DNA methylation of >485,000 CpG sites in maternal and cord blood. Results indicated a change in both DNA methylation and one-carbon metabolism indices over pregnancy (<12 week baseline to delivery) and a correlation between maternal and cord blood indices of both DNA methylation status and one-carbon metabolism. In addition, specific CpG sites that change over pregnancy associated with specific maternal or cord blood one carbon metabolism indices were identified, including an association between 5-methyltetrahydrofolate in cord blood and methylation changes in genes linked to vital neuronal and developmental processes. Overall, results of these studies identify specific changes in DNA methylation associated with obesity, pregnancy and alterations in folate status associated with these physiological conditions and provide evidence between potential links between alterations in maternal folate status and increased NTD risk.

Publications

• Type: Journal Articles Status: Published Year Published: 2017 Citation: Park, H. J., Bailey, L. B., Shade, D. C., Hausman, D. B., Hohos, N. M., Meagher, R. B., Kauwell, G. P., Lewis, R. D.,Smith, A. K. Distinctions in gene-specific changes in DNA methylation in response to folic acid supplementation between women of normal weight and obesity. Obes. Res. Clin. Prac. Jul 18. pii: S1871-403X(17)30062-5. doi: 10.1016/j.orcp.2017.06.004
• Type: Journal Articles Status: Awaiting Publication Year Published: 2017 Citation: Shade, D. C., Park, H. J., Hausman, D. B., Hohos, N. M., Meagher, R. B., Kauwell, G. P., Kilaru, V., Lewis, R. D.,Smith, A. K., Bailey, L. B. DNA methylation changes in whole blood and CD16+ neutrophils in response to chronic folic acid supplementation in women of childbearing age.. Int. J. Vitam. Nutr. Res. (In Press)
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Park, H. J., Knight, A. K., Fleming, J. M., Hausman, D. B., Caudill, M. A., Malysheva, O. V., Kauwell, G. P.A., Sokolow, A., Fisher, S., Smith, A. K., Bailey, L. Association Between One-carbon Metabolism Indices and DNA Methylation Status in Maternal and Cord Blood FASEB J 31:802.28.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Hohos, N., Smith, A. K., Kilaru, V., Park, H. J., Hausman, D., Bailey, L. B., Lewis, R. D., Meagher, R. B., Differential DNA Methylation between Obese and Normal Weight Women in CD4+ and CD8+ T cells, and CD16+ neutrophils FASEB J 30:431.1
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Park, H. J., Hausman, D., Fleming, J., Andersen, V., Rosa, G., Kauwell, G. P. A., Sokolow, A., Caudill, M. A., Malysheva, O., Fisher, S., Bailey, L. B., Folate and Choline Metabolite Changes in Maternal and Cord Blood in Response to Folic Acid Supplementation Dose During Pregnancy FASEB J 30:1172.10
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Fleming, J., Park, H. J., Hausman, D., Rosa, G., Andersen, V., Kauwell, G. P. A., Caudill, M. A., Malysheva, O., Fisher, S., Bailey, L. B., Unmetabolized Folic Acid Response to Folic Acid Supplementation Dose During Pregnancy FASEB J 30:1172.13
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Hausman, D., Holder, A., Park, H. J., Rosa, G., Andersen, V., Fleming, J., Kauwell, G. P. A., Caudill, M. A., Malysheva, O., Fisher, S., Bailey, L. B., Vitamin B12 Status and Infant Outcome in a Community-Based Folic Acid Intervention Study FASEB J 30:1172.7
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Bailey, L. B., Stover, P. J., McNulty, H., Fenech, M. F., Gregory, J. F. 3rd, Mills, J. L., Pfeiffer, C. M., Fazili, Z., Zhang, M.,Ueland, P. M., Molloy, A. M., Caudill, M. A., Shane, B., Berry, R. J., Bailey, R. L., Hausman, D. B., Raghavan, R., Raiten,D. J. (2015) Biomarkers of nutrition and development: Folate review. J. Nutr. 145:1636S-1680S.
• Type: Journal Articles Status: Published Year Published: 2012 Citation: Tinker, S.C., Hamner, H.C., Berry, R.J., Bailey, L.B., Pfeiffer, C.M. Does obesity modify the association of supplemental folic acid with folate status among nonpregnant women of childbearing age in the United States? Birth Defects Res A Clin Mol Teratol. 2012 Oct;94(10):749-55. doi: 10.1002/bdra.23024.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: da Silva, V. R., Hausman, D. B., Kauwell, G. P., Sokolow, A., Tackett, R. L., Bailey, L. B. Obesity affects short-term pharmacokinetics in women of childbearing age. Int. J. Obes. 37:1608-10.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Park, H. J., Shade, D., Hausman, D., Hohos, N., Meagher, R. B., Kauwell, G. P.A., Smith, A. K., Bailey, L. Gene-specific changes in DNA methylation in response to chronic folic acid supplementation in normal weight and obese women of child-bearing age FASEB J. 28: 817.3.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Shade, D., Park, H. J., Hausman, D., Hohos, N., Meagher, R. B., Kauwell, G. P.A., Smith, A. K., Bailey, L. Genome wide DNA methylation in whole blood and CD16+ cells in response to chronic folic acid supplementation in women of child-bearing age. FASEB J. 28:817.1
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Hohos, N., Park, H. J., Shade, D. C., Hausman, D., Meagher, R. B., Smith, A. K., Dellafera, M., Bailey, L., Baile, C. A. DNA methylation differences between normal weight and obese women of childbearing age. FASEB J. 28:1120.11
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Kauwell, G. P.A., Caudill, M., Hausman, D., Park, H. J., Shade, D., Malysheva, O. V., Bailey, L. Serum folate and whole blood global DNA methylation response to chronic folic acid supplementation in normal weight and obese women of child-bearing age FASEB J. 28: 817.2.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Anderson, A. K., Da Silva, V., Lewis, R. D., Rathbun, S. L., Hausman, D., Bailey, L. Response to short-term folic acid intake in relation to the body composition of women of reproductive age. FASEB J 27:lb249.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Da Silva, V., Hausman, D., Kauwell, G. P., Tackett, R. L., Rathbun, S. L., Bailey, L. Obesity affects short-term folic acid pharmacokinetics in women of childbearing age. FASEB J 27:246.2.
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: CURO Symposium. UGA Center for Undergraduate Research Opportunities. Neutrophil and granulocyte methylation in normal weight and obese individuals. Jenissa Gordon, Natalie Hohos, Deanna C. Shade, Dorothy B. Hausman, Gail P. A. Kauwell, Hea Jin Park Lynn B. Bailey, Athens, GA, April 4, 2016.
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: CURO Symposium. UGA Center for Undergraduate Research Opportunities. Blood Folate and Whole Blood Global Methylation Response to Folic Acid Supplementation Dose During Pregnancy. Joanna Yang, Dorothy B. Hausman, Jennifer Fleming, Gail P. A. Kauwell, Marie A. Caudill, Hea Jin Park, Lynn B. Bailey. Athens, GA, March 30, 2015.
• Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: CURO Symposium. UGA Center for Undergraduate Research Opportunities. Folic acid knowledge and use among participants of a community-based intervention study in pregnant women. Meagan Patterson, Lynn B. Bailey, Hea Jin Park and Dorothy B. Hausman. Athens, GA, March 30, 2015.
• Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: CURO Summer Symposium. UGA Center for Undergraduate Research Opportunities. Impact of gluten containing foods on folate intake of women of child bearing age. Jenissa Gordon, Dorothy B. Hausman, Lynn B. Bailey, Richard D. Lewis, Emma Laing, and Hea Jin Park. Athens, GA. July 31, 2015.
• Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Medical College of Georgia/University of Georgia Medical Partnership Student Research Symposium. Vitamin B12 status during pregnancy and infant outcome. Angela Holder, Hea Jin Park, Dorothy B. Hausman, Victoria Andersen, Gisselle Rosa, Jennifer Fleming, Susan Fisher and Lynn B. Bailey. Athens, GA. September 28, 2015.
• Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Georgia Academy of Nutrition and Dietetics Annual Conference and Exhibition. Folic acid supplementation during pregnancy: feasibility and implementation. Gisselle M. Rosa, Hea Jin Park, Dorothy B. Hausman, Victoria Andersen, Jennifer Fleming, Susan Fisher and Lynn B. Bailey. Athens, GA., March 18, 2015.
• Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Georgia Academy of Nutrition and Dietetics Annual Conference and Exhibition. Folic acid supplementation during pregnancy: baseline characteristics. Victoria Andersen, Hea Jin Park, Dorothy B. Hausman, Gisselle Rosa, Jennifer Fleming, Susan Fisher and Lynn B. Bailey. Athens, GA., March 18, 2015.
• Type: Conference Papers and Presentations Status: Other Year Published: 2014 Citation: CURO Symposium. UGA Center for Undergraduate Research Opportunities, Associations between body composition and serum folate concentrations in women of childbearing age: secondary analysis across three studies. Patterson, Meagan, Hausman, Dorothy. Athens, GA. March 31, 2014.
• Type: Theses/Dissertations Status: Published Year Published: 2016 Citation: Jennifer Fleming. Response of One-Carbon Biomarkers in Maternal and Cord Blood to Folic Acid Dose during Pregnancy. Masters Thesis. University of Georgia
• Type: Conference Papers and Presentations Status: Other Year Published: 2014 Citation: SEC Symposium: Prevention of Obesity: Overcoming a 21st Century Public Health Challenge. The Academic Initiative of the SEC, Atlanta, GA DNA Methylation Differences in CD16+ Neutrophils Between Normal Weight and Obese Women. Hohos, Natalie, Meagher, Richard B., Park, Hea Jin, Smith, Alicia K., Shade, Deanna, Hausman, Dorothy, Bailey. Atlanta, GA. September 22, 2014.
• Type: Conference Papers and Presentations Status: Other Year Published: 2014 Citation: SEC Symposium: Prevention of Obesity: Overcoming a 21st Century Public Health Challenge. The Academic Initiative of the SEC, Atlanta, GA Impact of obesity on folate metabolism and requirements of women of childbearing age. Hausman, Dorothy, Park, Hea Jin, Kauwell, Gail P A., Caudill, Marie, Meagher, Richard B., Smith, Alicia K., Anderson, Alex K., Lewis, Richard D., Birch, Leann, Rathbun, Stephen L., Fisher, Susan, Bailey, Lynn. Atlanta, GA. September 22, 2014.
• Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Obesity Week. Obesity Impacts the Serum Response to Chronic Folic Supplementation in Women of Child-Bearing Age. Deanna Shade, Hea Jin Park, Dorothy B. Hausman, D., Gail P. A. Kauwell, Andrew Sosolow, Stephen L. Rathbun, Lynn B. Bailey. Atlanta, GA. November, 2013.
• Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: CURO Symposium. UGA Center for Undergraduate Research Opportunities. Associations Between BMI and Folate Status of Normal Weight and Obese Women of Childbearing Age. Arielle Weekley, Dorothy B. Hausman and Lynn B. Bailey. Athens, GA. March 30, 2013.
• Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Southern Translational Education and Research, STaR Conference. Hea Jin Park, Dorothy B. Hausman, Vanessa da Silva, Gail P. A. Kauwell, Joseph Kindler, Richard D. Lewis, Lynn B. Bailey. Body composition impacts the serum response to acute folic acid supplementation in women of child bearing age. Augusta, GA September 26-27, 2013
• Type: Theses/Dissertations Status: Published Year Published: 2015 Citation: Victoria Andersen. Folic Acid Supplementation in Pregnancy: Maternal Outcomes. Masters Thesis. University of Georgia
• Type: Theses/Dissertations Status: Published Year Published: 2015 Citation: Gisselle Rosa. Folic Acid Supplementation during Pregnancy: Feasibility and Infant Outcomes. Masters Thesis. University of Georgia
• Type: Theses/Dissertations Status: Published Year Published: 2014 Citation: Deanna Shade. 4 DNA Methylation Changes in Whole Blood and CD16+ Neutrophils in Response to Chronic Folic Acid Supplementation in Women of Child-bearing Age. Masters Thesis. University of Georgia

Progress 10/01/15 to 09/30/16

Outputs
Target Audience:National and international folate research community, medical and allied health professionals, non-pregnant and pregnant women of child bearing age Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The research activites of this project provided professional development for a temporary assistant professor, three masters/dietetic intern students and several undergraduate majors in biology, nutrition science and dietetics. These individuals participated in data analysis and manuscript writing for the folate acid supplementation trial in women of childbearing age conducted in Year 2 of the project. They also participated in preparation of recruitment materials, supplement packages, and sampling packets for the recently launched folic acid supplementation trial in pregnant women conducted in collaboration with Athens Regional Midwifery Practice. The students were also responsible for contacting participants for obtaining Health Behavior Questionnaire data and gained expertise in the many other areas of clinical research such as processing of biological samples for biomarker and epigenetic analysis, supplement distribution and compliance monitoring and diet analysis using the ASA24 system, and data entry and statistical analysis. All trainees also attended bi-weekly journal club discussions of current literature related to folate status, folic acid supplementation, nutrient requirements during pregnancy and other topics pertinent to the research project. They also assisted in the preparation of publications for submission to peer-reviewed journals and/or prepared abstracts and research posters which they presented at local, regional or national conferences. How have the results been disseminated to communities of interest?The results have been disseminated through peer review publications and presentations at local and national svcientific conferences such as the CURO undergraduate research symposium and the FASEB conference. What do you plan to do during the next reporting period to accomplish the goals?Additonal epigenetic analyses will be performed to determined cell-specific changes in response to folic acid in maternal and infant blood DNA.

Impacts
What was accomplished under these goals? The epigenetic effect of supplemental folic acid on maternal and cord blood DNA was determined in a double-blind controlled intervention study. The impact of obesity on the epigneetic response to supploemental folic acid was determined.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Hea Jin Park, Dorothy B Hausman, Jennifer Fleming, Victoria Andersen, Gisselle Rosa, Gail Kauwell, Andrew Sokolow, Marie Caudill, Olga Malysheva, Susan Fisher, Lynn B Bailey. Folate and Choline Metabolite Changes in Maternal and Cord Blood in Response to Folic Acid Supplementation Dose During Pregnancy FASEB J April 2016 30:1172.10
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Jennifer Fleming, Hea Jin Park, Dorothy B Hausman, Gisselle Rosa, Victoria Andersen, Gail Kauwell, Marie Caudill, Olga Malysheva, Susan Fisher, and Lynn B Bailey. Unmetabolized Folic Acid Response to Folic Acid Supplementation Dose During Pregnancy FASEB J April 2016 30:1172.13
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Dorothy B Hausman, Angela Holder, Hea Jin Park, Gisselle Rosa, Victoria Andersen, Jennifer Fleming, Gail Kauwell, Marie Caudill, Olga Malysheva, Susan Fisher, and Lynn B Bailey. Unmetabolized Folic Acid Response to Folic Acid Supplementation Dose During Pregnancy FASEB J April 2016 30:1172.13
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Natalie M Hohos, Alicia K Smith, Varun Kilaru, Hea Jin Park, Dorothy B Hausman, Lynn B Bailey, Richard D Lewis, and Richard B Meagher. Differential DNA Methylation between Obese and Normal Weight Women in CD4+ and CD8+ T cells, and CD16+ neutrophils FASEB J April 2016 30:431.1
• Type: Journal Articles Status: Published Year Published: 2016 Citation: An epigenetic clock for gestational age at birth based on blood methylation data. Knight AK et al. Genome Biol. 2016 Oct 7;17(1):206.

Progress 10/01/14 to 09/30/15

Outputs
Target Audience:National and international folate research community, medical and allied health professionals, non-pregnant and pregnant women of child bearing age ? Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The research activities of this project provided opportunities for professional development for a temporary assistant professor, three masters/dietetic intern students and several undergraduate majors in biology, nutrition science and dietetics. In addition, a first-year medical student from the How have the results been disseminated to communities of interest?The results have been disseminated through peer review publications and presentations at local and state-wide scientific conferences such as the CURO undergraduate research symposium, MCG/UGA Medical Partnership Student Research Symposium and the Georgia Academy of Nutrition and Dietetics Annual Conference. ? What do you plan to do during the next reporting period to accomplish the goals?Data and statistical analysis for the folic acid supplementation trial in pregnant women is on-going. Specifically, analysis of genome-wide and gene-specific response to the folic acid supplementation is being conducted in collaboration with Dr. Alicia Smith at Emory University. Several abstracts related to the project were submitted late 2015 (to be reported in next reporting period) or early in 2016 and will be presented at the upcoming CURO2016 and EB2016 conferences. It is anticipated that several manuscripts based on these results of this trial will be drafted and submitted in 2016. ?

Impacts
What was accomplished under these goals? Recent data indicates that almost 60% of women of reproductive age are overweight or obese and that obese women are at higher risk for having a neural tube defect (NTD) affected pregnancy. Current recommendations for optimal intake of folate for women of this age group are based on periconceptional levels shown to be effective for NTD risk reduction, but do not take body weight into consideration. The purpose of this project is to evaluate the impact of obesity on folate metabolism and requirements. The project has four objectives, two of which were addressed during this project period: 1) To determine response to folic acid supplementation in normal weight and obese women and 2) To explore epigenetic biomarkers of folate status. Research during this project period focused on the folic acid supplementation dose response study in pregnant women conducted in collaboration with Athens Regional Midwifery Practice. This is a double-blind randomized controlled trial in healthy pregnant women (BMI 18.5 - 29.9 kg/m2, age 18 - 40 yr) receiving prenatal care at Athens Regional Midwifery clinic (ARMC). The primary goal of the study is to determine relative changes in blood folate and DNA methylation levels in response to two different supplemental doses of folic acid one of which provides the Institute of Medicine's recommended folate intake (400 mcg/d folic acid) for pregnant women and the second which provides a higher dose (800 mcg/d folic acid) as routinely taken by pregnant women in over-the-counter prenatal supplements.

Publications

• Type: Journal Articles Status: Published Year Published: 2015 Citation: Bailey, L. B., Stover, P. J., McNulty, H., Fenech, M. F., Gregory, J. F. 3rd, Mills, J. L., Pfeiffer, C. M., Fazili, Z., Zhang, M., Ueland, P. M., Molloy, A. M., Caudill, M. A., Shane, B., Berry, R. J., Bailey, R. L., Hausman, D. B., Raghavan, R., Raiten, D. J. (2015) Biomarkers of nutrition and development: Folate review. J. Nutr. 1636S-1680S.
• Type: Journal Articles Status: Under Review Year Published: 2016 Citation: Park, H. J., Bailey, L. B., Shade, D. C., Hausman, D. B., Hohos, N. M., Meagher, R. B., Kauwell, G. P., Lewis, R. D., Smith, A. K. Distinctions in gene-specific changes in DNA methylation in response to chronic folic acid supplementation between normal weight and obese women of childbearing age. Clinical Epigenetics (under review)
• Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Georgia Academy of Nutrition and Dietetics Annual Conference and Exhibition. Folic acid supplementation during pregnancy: feasibility and implementation. Gisselle M. Rosa, Hea Jin Park, Dorothy B. Hausman, Victoria Andersen, Jennifer Fleming, Susan Fisher and Lynn B. Bailey. Athens, GA., March 18, 2015.
• Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Georgia Academy of Nutrition and Dietetics Annual Conference and Exhibition. Folic acid supplementation during pregnancy: baseline characteristics. Victoria Andersen, Hea Jin Park, Dorothy B. Hausman, Gisselle Rosa, Jennifer Fleming, Susan Fisher and Lynn B. Bailey. Athens, GA., March 18, 2015.
• Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: CURO Summer Symposium. UGA Center for Undergraduate Research Opportunities. Impact of gluten containing foods on folate intake of women of child bearing age. Jenissa Gordon, Dorothy B. Hausman, Lynn B. Bailey, Richard D. Lewis, Emma Laing, Hea Jin Park. Athens, GA. July 31, 2015.
• Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Medical College of Georgia/University of Georgia Medical Partnership Student Research Symposium. Vitamin B12 status during pregnancy and infant outcome. Angela Holder, Hea Jin Park, Dorothy B. Hausman, Victoria Andersen, Gisselle Rosa, Jennifer Fleming, Susan Fisher and Lynn B. Bailey. Athens, GA. September 28, 2015.
• Type: Theses/Dissertations Status: Published Year Published: 2015 Citation: Gisselle Rosa. 2015. Folic Acid Supplementation during Pregnancy: Feasibility and Infant Outcomes. Masters Thesis. University of Georgia
• Type: Theses/Dissertations Status: Published Year Published: 2015 Citation: Victoria Andersen. 2015. Folic Acid Supplementation in Pregnancy: Maternal Outcomes. Masters Thesis. University of Georgia
• Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: CURO Symposium. UGA Center for Undergraduate Research Opportunities. Factors influencing folate status in Hispanic women. Courtney Alvis, Dorothy B. Hausman, Hea Jin Park and Lynn B. Bailey. Athens, GA, March 30, 2015.
• Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: CURO Symposium. UGA Center for Undergraduate Research Opportunities. Folic acid knowledge and use among participants of a community-based intervention study in pregnant women. Meagan Patterson, Lynn B. Bailey, Hea Jin ark and Dorothy B. Hausman. Athens, GA, March 30, 2015.

Progress 10/01/13 to 09/30/14

Outputs
Target Audience:National and international folate research community, medical and allied health professionals, non-pregnant and pregnant women of child bearing age Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The research activities of this project provided opportunities for professional development for a temporary assistant professor, three masters/dietetic intern students and several undergraduate majors in biology, nutrition science and dietetics. These individuals participated in data analysis and manuscript writing for the folate acid supplementation trial in women of childbearing age conducted in Year 2 of the project. They also participated in preparation of recruitment materials, supplement packages, and sampling packets for the recently launched folic acid supplementation trial in pregnant women conducted in collaboration with Athens Regional Midwifery Practice. The students were also responsible for contacting participants for obtaining Health Behavior Questionnaire data and gained expertise in the many other areas of clinical research such as processing of biological samples for biomarker and epigenetic analysis, supplement distribution and compliance monitoring and diet analysis using the ASA24 system, and data entry and statistical analysis. All trainees also attended bi-weekly journal club discussions of current literature related to folate status, folic acid supplementation, nutrient requirements during pregnancy and other topics pertinent to the research project. They also assisted in the preparation of publications for submission to peer-reviewed journals and/or prepared abstracts and research posters which they presented at local, regional or national conferences. How have the results been disseminated to communities of interest?The results have been disseminated through a peer review publications and presentations at local, regional and national scientific conferences such as Experimental Biology (annual meeting of American Society for Nutrition) and the SEC Symposium: Prevention of Obesity: Overcoming a 21st Century Public Health Challenge. What do you plan to do during the next reporting period to accomplish the goals?The folic acid supplementation trial in pregnant women is on-going. We anticipate closing recruitment at the end of 2014. Samples will continue to be collected at 28 and 36 weeks of pregnancy and from the mothers and infants at delivery. Stored samples will be shipped for biomarker and DNA methylation analysis immediately following the last delivery which is projected to be in late July/early August 2015.

Impacts
What was accomplished under these goals? Recent data indicates that almost 60% of women of reproductive age are overweight or obese and that obese women are at higher risk for having a neural tube defect (NTD) affected pregnancy. Current recommendations for optimal intake of folate for women of this age group are based on periconceptional levels shown to be effective for NTD risk reduction, but do not take body weight into consideration. The purpose of this project is to evaluate the impact of obesity on folate metabolism and requirements. The project has four objectives, two of which were addressed during this project period: 1) To determine response to folic acid supplementation in normal weight and obese women and 2) To explore epigenetic biomarkers of folate status. Research during this project period focused on two clinical trials (a) the folic acid supplementation study in normal weight and obese women of childbearing age [conducted in the previous project period] for which considerable efforts were devoted to sample and data analysis and manuscript writing and (b) a folic acid supplementation dose response study in pregnant women initiated in collaboration with Athens Regional Midwifery Practice. For the first of these studies, genome wide and gene-specific DNA methylation response to folic acid supplementation was determined in whole blood and CD16+ cells from 12 normal weight. DNA methylation patterns measured across 485,000 CpG sites throughout the genome indicated a change in response to folic acid supplementation in 7839 and 8317 CpG sites (p<0.05) in whole blood and CD16+ neutrophils, respectively and 6 obese women. This suggests that the genome-wide DNA methylation response to chronic folic acid supplementation is different between whole blood and CD16+ neutrophils and that a single white blood cell type may function as a more specific epigenetic reporter of folate status than whole blood. In additional analysis, changes in DNA methylation in response to the folic acid supplementation were compared in CD16+ cells isolated from 12 normal weight and 6 obese women. Examination of DNA methylation of 2,098 CpG sites in 91 genes related to NTD risk and folate metabolism indicated a substantial difference in response between the normal weight and obese women suggesting that differences in DNA methylation and response to supplementation may lead to increased NTD risk and abnormal folate metabolism in obesity. The second study, which was initiated during this project period, is a double-blind randomized controlled trial in healthy pregnant women (BMI 18.5 - 29.9 kg/m2, age 18 - 40 yr) receiving prenatal care at Athens Regional Midwifery clinic (ARMC). The primary goal of the study is to determine relative changes in blood folate and DNA methylation levels in response to two different supplemental doses of folic acid one of which provides the Institute of Medicine's recommended folate intake (400 mcg/d folic acid) for pregnant women and the second which provides a higher dose (800 mcg/d folic acid) as routinely taken by pregnant women in over-the-counter prenatal supplements. After establishing collaboration, developing protocols and receipt of IRB approval and both UGA and Athens Regional Medical Center, midwives and other project personnel were trained in study protocols. Pregnant women are being recruited at their first prenatal visit (< 12 weeks) and baseline blood samples for determination of folate status and DNA methylation are taken at that time. Women are then randomly assigned to take supplements containing either 400 or 800 mcg/d folic acid throughout pregnancy. Additional blood samples will be taken at 28 and 36 weeks gestation and from the mother and infant (cord blood) at delivery. Recruitment for the study began in mid-July 2014 and 24 participants have been recruited to-date (9/30/14). Twenty-eight week sampling will begin in late November and the first deliveries are expected in mid-February. Recruitment will continue through the end of 2014 with the hopes of enrolling at least 50 women. Biological samples are being stored at -80?C and will be analyzed upon completion of the project. It is anticipated that this investigation will provide new evidence that will link and compare observed changes in DNA methylation of specific genes associated with birth defects and chronic disease in response to these two prenatal folic acid doses and characterize the impact of maternal obesity on the epigenomic response.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: SEC Symposium: Prevention of Obesity: Overcoming a 21st Century Public Health Challenge. The Academic Initiative of the SEC, Atlanta, GA Impact of obesity on folate metabolism and requirements of women of childbearing age. Hausman, Dorothy, Park, Hea Jin, Kauwell, Gail P A., Caudill, Marie, Meagher, Richard B., Smith, Alicia K., Anderson, Alex K., Lewis, Richard D., Birch, Leann, Rathbun, Stephen L., Fisher, Susan, Bailey, Lynn. September 22, 2014.
• Type: Journal Articles Status: Under Review Year Published: 2015 Citation: Shade, D. C., Park, H. J., Hausman, D., Hohos, N., Meagher, R. B., Kauwell, G. P. A., Kilaru, V., Lewis, R. D., Smith, A. K., Bailey, L. (2014). DNA methylation changes in whole blood and CD16+ neutrophils in response to chronic folic acid supplementation in women of childbearing age. PLoS ONE. (under review)
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Park, H. J., Shade, D., Hausman, D., Hohos, N., Meagher, R. B., Kauwell, G. P.A., Smith, A. K., Bailey, L. (2014). Gene-specific changes in DNA methylation in response to chronic folic acid supplementation in normal weight and obese women of child-bearing age (vol. 28, pp. 817.3). FASEB J.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Shade, D., Park, H. J., Hausman, D., Hohos, N., Meagher, R. B., Kauwell, G. P.A., Smith, A. K., Bailey, L. (2014). Genome wide DNA methylation in whole blood and CD16+ cells in response to chronic folic acid supplementation in women of child-bearing age (vol. 28, pp. 817.1). FASEB J.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Hohos, N., Park, H. J., Shade, D. C., Hausman, D., Meagher, R. B., Smith, A. K., Dellafera, M., Bailey, L., Baile, C. A. (2014). DNA methylation differences between normal weight and obese women of childbearing age (vol. 28, pp. 1120.11). FASEB J.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Kauwell, G. P.A., Caudill, M., Hausman, D., Park, H. J., Shade, D., Malysheva, O. V., Bailey, L. (2014). Serum folate and whole blood global DNA methylation response to chronic folic acid supplementation in normal weight and obese women of child-bearing age (vol. 28, pp. 817.2). FASEB J.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: 2014 CURO Symposium. UGA Center for Undergraduate Research Opportunities, Athens, GA Associations between body composition and serum folate concentrations in women of childbearing age: secondary analysis across three studies. Patterson, Meagan, Hausman, Dorothy. March 31, 2014.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: SEC Symposium: Prevention of Obesity: Overcoming a 21st Century Public Health Challenge. The Academic Initiative of the SEC, Atlanta, GA DNA Methylation Differences in CD16+ Neutrophils Between Normal Weight and Obese Women. Hohos, Natalie, Meagher, Richard B., Park, Hea Jin, Smith, Alicia K., Shade, Deanna, Hausman, Dorothy, Bailey, Lynn, Bailey, Lynn. September 22, 2014.
• Type: Theses/Dissertations Status: Published Year Published: 2014 Citation: Deanna Shade. 2014. DNA Methylation Changes in Whole Blood and CD16+ Neutrophils in Response to Chronic Folic Acid Supplementation in Women of Child-bearing Age. Masters Thesis. University of Georgia

Progress 01/01/13 to 09/30/13

Outputs
Target Audience:National and international folate research community, medical and allied health professionals, non-pregnant and pregnant women of child bearing age Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The research activities of this project provided opportunities for professional development for one post-doctoral associate, a temporary assistant professor, one masters/dietetic intern student and several undergraduate majors in biology, nutrition science and dietetics. These individuals participated in the conduct of a folate acid supplementation trial in women of childbearing age and gained expertise in the many areas such as subject recruitment and retention, processing of biological samples for biomarker and epigenetic analysis, supplement distribution and compliance monitoring, diet analysis using the ASA24 system, and anthropometric and body composition analysis. All trainees also attended bi-weekly journal club discussions of current literature related to folate status, folic acid supplementation, DNA methylation and other topics pertinent to the research project. They also assisted in the preparation of publications for submission to peer-reviewed journals and/or prepared abstracts and research posters which they presented at local, regional or national conferences. How have the results been disseminated to communities of interest?The results have been disseminated through a peer review publications and presentations at local, regional and national scientific conferences such as Obesity Week and Experimental Biology (annual meeting of American Society for Nutrition). What do you plan to do during the next reporting period to accomplish the goals?Beginning in Fall 2013, we hope to establish collaboration with the Athens Regional Midwifery Practice with the intent of initiating a folic acid supplementation trial in pregnant women in Spring 2014.

Impacts
What was accomplished under these goals? Recent data indicates that almost 60% of women of reproductive age are overweight or obese and that obese women are at higher risk for having a neural tube defect (NTD) affected pregnancy. Current recommendations for optimal intake of folate for women of this age group are based on periconceptional levels shown to be effective for NTD risk reduction, but do not take body weight into consideration. The purpose of this project is to evaluate the impact of obesity on folate metabolism and requirements. The project has four objectives, two of which were addressed during this project period: 1) To determine response to folic acid supplementation in normal weight and obese women and 2) To explore epigenetic biomarkers of folate status. Upon receipt of IRB approval, a folic acid supplementation study was initiated in normal weight and obese women recruited from the University of Georgia and surrounding community. In the initial phase of the study 50 nonpregnant women age 18-40 yr were screened for baseline folate status and MTHFR genotype. Fourteen normal weight (BMI 18.5-24.9 kg/m2) and eight obese (BMI > 30 kg/m2) women with serum folate <60 nmol/L and the CC or CT genotype at MTHFR 677 entered the second phase of the study, an eight week supplementation trial with 800 ug/day folic acid. Blood samples were collected at baseline and after 4 and 8 weeks supplementation for determination of biomarkers of folate status and DNA methylation response to the supplementation. The clinical phase of the study was initiated in April 2013 and completed by early August. Initial results indicate that serum folate concentration at baseline was lower in obese than in normal weight women (p=.015) and inversely related to BMI (r=.551, p=0.012). In the normal weight group, serum folate concentration consistently increased with folic acid supplementation from baseline (39.6±2.6 nmol/L) to four (57.4±3.8 nmol/L) and eight (71.5±2.8 nmol/L) weeks. In the obese group, serum folate concentration increased from baseline (28.4±2.9 nmol/L) to four weeks (57.5±7.1 nmol/L; non-significant vs. normal weight) but plateaued by eight weeks (59. 3±4.1 nmol/L; p=.021 vs. normal weight). The findings of lower baseline folate concentrations and a plateaued response to chronic folic acid supplementation in obese as compared with normal weight women provides additional evidence of a negative impact of obesity on folate metabolism and additional support for a BMI-adjusted folic acid intake recommendation. Global, genome-wide and gene specific DNA methylation response is currently being analyzed in whole blood and CD16+ cells to determine if there are differences in baseline methylation status and methylation response to folic acid supplementation between normal weight and obese women of childbearing age.

Publications

• Type: Journal Articles Status: Published Year Published: 2013 Citation: Da Silva, V., Hausman, D., Kauwell, G., Sokolow, A., Tackett, R. L., Rathbun, S. L., Bailey, L. (2013). Obesity affects short-term folic acid pharmacokinetics in women of childbearing age. International Journal of Obesity. 37:1608-10
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2013 Citation: Shade, D., Park, H. J., Hausman, D., Kauwell, G. P.A., Sosolow, A., Rathbun, S. L., Bailey, L. (2013). Obesity Impacts the Serum Response to Chronic Folic Supplementation in Women of Child-Bearing Age. Obesity Week 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Anderson, A. K., Da Silva, V., Lewis, R. D., Rathbun, S. L., Hausman, D., Bailey, L. (2013). Response to short-term folic acid intake in relation to the body composition of women of reproductive age. FASEB J April 9,2013 27:lb249.
• Type: Conference Papers and Presentations Status: Awaiting Publication Year Published: 2013 Citation: Da Silva, V., Hausman, D., Kauwell, G. P., Tackett, R. L., Rathbun, S. L., Bailey, L. (2013). Obesity affects short-term folic acid pharmacokinetics in women of childbearing age. FASEB J April 9, 2013 27:246.2.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Southern Translational Education and Research STaR Conference 2013. UGA Pharmacy; GRU Medical College, Augusta, GA Body composition impacts the serum response to acute folic acid supplementation in women of child bearing age. Park, Hea Jin, Hausman, Dorothy, Da Silva, Vanessa, Kauwell, Gail P.A., Kindler, Joseph, Lewis, Richard D., Bailey, Lynn. September 27, 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Southern Translational Education and Research STaR Conference 2013. UGA Pharmacy; GRU Medical College, Augusta, GA Obesity impacts the serum response to chronic folic acid supplementation in women of child-bearing age. Shade, Deanna, Park, Hea Jin, Hausman, Dorothy, Kauwell, Gail P.A., Sokolow, Andrew, Bailey, Lynn. September 27, 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: CURO 2013. UGA Center for Undergraduate Research Opportunities, Athens, GA Associations between BMI and Folate Status of Normal Weight and Obese Women of Childbearing Age. Weekley, Arielle, Hausman, Dorothy, Bailey, Lynn. April 1, 2013.

Progress 07/01/12 to 12/31/12

Outputs
OUTPUTS: The purpose of this project is to evaluate the impact of obesity on folate metabolism and requirements. Specifically, the adequacy of the current RDA of 400 mcg/d DFE for obese women is being addressed. The project has four objectives, two of which were addressed in this initial project period: 1) To determine short-term response in serum folate levels after a single dose of folic acid in normal weight and obese women and 2) To investigate the associations between the total adipose tissue, as well as body fat distribution, and folate status and clearance from serum. This was accomplished by means of a folic acid pharmacokinetics study conducted in normal weight (BMI 18.5 - 24.9 kg/m2) and obese (BMI > 30.0 kg/m2) women of childbearing age (18-35 yr; n=16 each). Non-pregnant Caucasian women meeting specific inclusion/exclusion criteria with regard to age, body weight status, absence of chronic disease, acute illness or use of prescription drugs, antibiotics (recent) or dietary supplements were recruited into the study. Following a baseline blood draw for determination of serum and red blood folate, the women were administered a single oral 400 mcg dose of folic acid and blood samples were collected over the subsequent 10 hour period to evaluate the serum folate response. Serum folate response was calculated by subtracting the baseline concentrations from the folate value at each subsequent time point. Individual response-time curves were generated and area-under-the-curve (AUC) was calculated using the trapezoidal rule. Maximum concentration response (Cmax) and time to peak concentration (tmax) were derived from the individual response curves. To evaluate the association between folate status and clearance and adiposity, percent body fat and lean mass of the participants was determined on the day of the study using the Bod Pod body composition tracking system and verified by dual-energy X-ray absorptiometry. As an indicator of typical folate and total food intake, participants completed the Automated Self-Administered 24-hour dietary recall (ASA24) including recalls for three separate days (two weekdays and one weekend day). Statistical analyses were performed using SigmaPlot (Systat) and SAS. Differences between biochemical and pharmacokinetic parameters among the normal weight and the obese groups were analyzed using two-tailed t-tests. A linear mixed-effects model was used to describe differences between response-time curves between groups. Subjects were treated as random effects and an exponential correlation function was used to describe dependence among repeated observations within subjects. PARTICIPANTS: Lynn B. Bailey, Ph.D., Principal Investigator - Department of Foods and Nutrition, University of Georgia Vanessa DaSilva, Ph.D., Study Co-ordinator, Postdoctoral Associate - professional development in conduct of clinical studies -- Department of Foods and Nutrition, University of Georgia Dorothy B. Hausman, Ph.D., Co-Investigator - Department of Foods and Nutrition, University of Georgia Gail P. Kauwell, Ph.D., Collaborator -- Food Science and Human Nutrition Department, University of Florida TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
A total of 16 obese and 16 normal weight women participated in the folic acid pharmacokinetics study. The two groups did not differ in average age or height. Baseline serum folate concentrations were lower (29% ; P < 0.01) in the obese group compared to the normal weight women. Conversely, the obese women had higher (14%: P < 0.05) RBC folate concentrations. There were no observed differences between groups in either self-reported total folate intake or in total folic acid intake from fortified foods (n=14 obese, n=11 normal weight). Total serum folate area under the curve (AUC) in response to the folic acid dose (0 - 10 h) was similar in both groups. However, obese and normal weight women had different overall serum responses to the folic acid dose administered (p<0.001). The maximum concentration response (Cmax) was 34% lower in the obese women (23.4 + 7.1 nmol/L) compared to the normal weight group (35.4 + 12.2 nmol/L; p<0.005). The average time to peak concentration ( tmax) for the normal weight women was 2.3 h, with all 16 participants reaching Cmax within the first 3 h. On the other hand, the average tmax for the obese group was 3.7 h (range: 1.5 - 9 h). We therefore calculated AUC0 -3 h, and observed a significantly higher AUC in the normal weight women compared to the obese group. This suggests a more rapid cellular folate uptake in the obese group, potentially related to increased adiposity. Accordingly, we investigated the relationship between the short term folate pharmacokinetic response and adiposity, defined dichotomously as normal < 32% body fat (BF) or high > 32% BF. In these analyses, there was a significant difference between % BF and baseline RBC folate (p < 0.05) with participants with high % BF having higher folate levels. In addition, serum folate response differed between normal % BF and high % BF, with a higher post dose peak observed for participants with normal % BF. Together, our findings indicate that the rate of absorption and distribution of folate differs between normal weight and obese women of child bearing age and is specifically impacted by adiposity. Impact: Recent data indicates that almost 60% of women of reproductive age are overweight or obese and that obese women are at higher risk for having a neural tube defect (NTD) affected pregnancy. Current recommendations for optimal intake of folate for women of this age group are based on periconceptional levels shown to be effective for NTD risk reduction, but do not take body weight into consideration. The findings of this short-term pharmacokinetic study are the first to provide evidence of a negative impact of obesity on the metabolic response to the current recommended intake of folic acid for NTD risk reduction. Further studies are required to clarify the relationship between obesity, adiposity and long-term folate pharmacokinetics. Such investigations would provide definitive evidence of the metabolic fate of folate in obese women and provide additional support for revised BMI- or adiposity-adjusted folate/folic acid intake recommendations for women of reproductive age.

Publications

• Publications: Da Silva, V., Hausman, D., Kauwell, G., Sokolow, A., Tackett, R. L., Rathbun, S. L., Bailey, L. 2013. Obesity affects short-term folic acid pharmacokinetics in women of childbearing age. International Journal of Obesity (In Press).
• Anderson, A. K., DaSilva, V. R., Lewis, R. D., Rathbun, S., Hausman, D. B., Bailey, L. B. 2013. Response to short-term folic acid intake in relation to the body composition of women of reproductive age. FASEB J (In Press).