Source: CORNELL UNIVERSITY submitted to NRP
HUMAN NUTRIENT REQUIREMENTS IN OBESITY, PREGNANCY AND CHILD DEVELOPMENT
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
SPECIAL GRANT
Reporting Frequency
Annual
Accession No.
0221596
Grant No.
2010-34324-20769
Cumulative Award Amt.
$350,862.00
Proposal No.
2010-01470
Multistate No.
(N/A)
Project Start Date
Jul 1, 2010
Project End Date
Jun 30, 2012
Grant Year
2010
Program Code
[DJ]- Human Nutrition, NY
Recipient Organization
CORNELL UNIVERSITY
(N/A)
ITHACA,NY 14853
Performing Department
Division of Nutritional Sciences
Non Technical Summary
Restructuring the wholesomeness of the food supply, and promoting healthy dietary habits and physical activity, increasingly are identified by the USDA as major public health objectives to secure longer lives and better quality of living throughout the life cycle. To achieve these goals, dietary recommendations must be increasingly grounded in an understanding of human nutrient dynamics and thereby result in more accurate and scientifically informed nutrition and agriculture policies for public health benefit. Targeted manipulation of the food supply will benefit western societies that seek to optimize human nutrition throughout the life cycle and potentially replace mass fortification initiatives which may place some individuals at risk. The overarching objective of this proposal is to increase our fundamental knowledge of human nutrition. These objectives support and advance the national goals of NIFA. Cornell University is uniquely positioned to address these research priorities because of its ongoing commitment of resources (facilities, faculty recruitments) that support a university-wide life-sciences initiative. This unique and targeted allocation of university resources and the associated academic environment are value-added to funds provided by this proposal, and together enable the development of nutritional genomics and human metabolic research facilities and permits Cornell scientists to address research questions in the emerging field of Human Nutrition and Human Genomics that will directly impact food and nutrition policy.
Animal Health Component
90%
Research Effort Categories
Basic
10%
Applied
90%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
7026010101070%
7026010108010%
7246010101010%
7246010108010%
Knowledge Area
724 - Healthy Lifestyle; 702 - Requirements and Function of Nutrients and Other Food Components;

Subject Of Investigation
6010 - Individuals;

Field Of Science
1010 - Nutrition and metabolism; 1080 - Genetics;
Goals / Objectives
The goals of this project are to initiate new human metabolic studies that address the role of iron, vitamin K and choline nutrition, as well as endocrine-related obesity, in human health. The proposed studies use of state-of-the-art molecular biology, imaging and stable isotope approaches to understand human nutrient dynamics at the whole body and cellular level in healthy humans. The research projects outlined in this proposal will: 1) investigate the effects of obesity on iron status in vulnerable pregnant populations ; 2) elucidate the metabolic signals that regulate abnormal follicle growth kinetics and anovulation in overweight/obese women ; 3) characteriza a new and novel deprenylation pathway of vitamin K metabolism and its stimulation by warfarin; and 4) determine the functional effects of varied choline intake during pregnancy on markers of DNA integrity/damage in lymphocytes and placentas from pregnant and nonpregnant women. The experiments will be conducted Cornell's Human Metabolic Research Facility (HMRU) at Cornell University. Results from these human metabolic studies are anticipated to support the determination of dietary recommendations. These objectives support and advance the national goals of CSREES that include sustaining a: 1) healthy, well-nourished population, 2) safe, secure food and fiber system, 3) agricultural production system that is highly competitive in the global economy, including the training of future professionals and scientists.
Project Methods
Project 1: The pilot studies will focus on 2 vulnerable populations; pregnant adolescents and women carrying multiple fetuses. Preliminary data will be obtained on novel approaches to measure plasma volume using new innovative stable isotopic mass spectrometric approaches and clinical studies will be undertaken to assess the impact of maternal obesity on iron status and absorption in these groups. Data from these studies will be used for subsequent grant applications in this area of research and results generated will impact public health recommendations and nutritional guidelines for pregnant women and adolescents. Project 2: The overall objective of this study is to elucidate the metabolic signals that regulate abnormal follicle growth kinetics and anovulation in overweight/obese women. The following Hypotheses will be tested in the pilot studies proposed: 1. Obesity-related amenorrhea reflects abnormal follicle wave development in women of reproductive age. 2. Overweight/obese women with abnormal follicle dynamics demonstrate a distinct metabolic profile compared to age- and body weight-matched controls with normal follicle waves and regular ovulatory menstrual cycles. 3. Resumption of follicle wave development occurs in overweight/obese women engaged in diet and exercise regimens that normalize metabolic signals. The long-term project goals are to develop nutritional, lifestyle and pharmaceutical regimens that normalize follicle wave dynamics and improve health outcomes in women of reproductive age. Project 3: Characterize the vitamin K deprenylation pathway, including the gene/gene product involved. and the mechanism by which warfarin stimulates vitamin K deprenylation. Project 4: Markers of DNA integrity/damage will be assessed in lymphocytes from pregnant and nonpregnant women consuming two doses of choline intake as well as in placental tissue from the pregnant study participants. It is hypothesized that higher choline intake will result in lower DNA damage. The following Hypotheses will be assessed in the pilot studies proposed: (i) that varied choline intake will influence circulating concentrations of pro-inflammatory markers in third trimester pregnant women and in nonpregnant control women. (ii) that varied choline intake will result in differential gene expression in placental tissue and/or circulating lymphocytes. (iii) that varied choline intake will alter markers of DNA damage in placental tissue and/or circulating lymphocytes. This hypothesis will be tested by comparing DNA damage, as measured by the Comet Assay, in placental tissue and circulating lymphocytes between women consuming 450 versus 900 mg/d choline.

Progress 07/01/10 to 06/30/12

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? All of the goals have been completed sucessfully as outlined in the progress reports and the results published in scientific, peer-reviewed journals.

Publications

  • Type: Journal Articles Status: Published Year Published: 2013 Citation: " Lujan ME, Peppin AK, Brooks ED, Reines JK, Jarrett BY, Pierson RA, Muhn N, Haider E, Chizen DR. Revised ultrasound criteria for polycystic ovary syndrome: reliable thresholds for elevated follicle population and ovarian volume. Hum Reprod. 2013; 28(5):1361-8.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2013 Citation: " Clark NM, Podolski AJ, Chizen DR, Pierson RA, Lehotay DC, Lujan ME. Prevalence of polycystic ovary syndrome (PCOS) phenotypes using updated criteria for polycystic ovarian morphology. Under review.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2013 Citation: " Christ J, Brooks ED, Reines J, Lujan ME. Heterogeneity in antral follicle populations relate to degree of reproductive and metabolic disturbance in women with polycystic ovary syndrome. Under Review.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2013 Citation: " Christ J, Brooks ED, Jarrett BY, Lujan ME. Assessments of the ovarian stroma do not improve diagnostic potential of follicle counts to predict polycystic ovary syndrome. Under review.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2013 Citation: " Jarrett BY, Pawlak AC, Lujan ME. Characterization of follicle waves during natural cycles using 3D ultrasonography and semi-automated follicle counting software. In preparation
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: " West AA, Yan J, Perry CA, Jiang X, Malysheva OV, Innis SE, Caudill MA. Choline intake influences phosphatidylcholine docosahexaenoic acid enrichment in nonpregnant women but not third-trimester pregnant women. Am J Clin Nutr. 2013 97(4):718-27. PMID:23446897[PubMed - in process]
  • Type: Journal Articles Status: Accepted Year Published: 2013 Citation: " Jiang X, Bar HY, Yan J, Jones S, Brannon PM, West AA, Perry CA, Malysheva OV, Pressman E, Devapatla S, Vermeylen F, Wells MT, Caudill MA. A higher maternal choline intake decreases the anti-angiogenic factor fms-like tyrosine kinase-1 (sFLT1) in both the placenta and maternal circulation. FASEB J. 2013 27(3):1245-53. PMID:23195033[PubMed - in process]
  • Type: Journal Articles Status: Accepted Year Published: 2012 Citation: " Jiang X, Bar HY, Yan J, West AA, Perry CA, Malysheva OV, Devapatla S, Pressman E, Vermeylen F, Wells MT, Caudill MA. Human pregnancy induces transcriptional activation of the peripheral innate immune system and increases oxidative DNA damage. PLoS ONE 7 2012 (11): e46736. doi:10.1371/journal.pone.0046736. PMID:23133592 PubMed - in process] PMCID:PMC3487782
  • Type: Journal Articles Status: Accepted Year Published: 2012 Citation: " West AA, Jian J, Perry CA, Jiang X, Malysheva OV, Caudill MA. Folate status response to a controlled folate intake among nonpregnant, pregnant, and lactating women. Am J Clin Nutr 2012;96:789800. PMID:22932279
  • Type: Journal Articles Status: Published Year Published: 2012 Citation: " Jiang X, Yan J, West AA, Perry CA, Malysheva OV, Devapatla S, Pressman E, Vermeylen F, Caudill MA. Maternal choline intake alters the epigenetic state of fetal cortisol regulating genes in humans. FASEB J. 2012 Aug;26(8):3563-74. PMID:22549509
  • Type: Journal Articles Status: Accepted Year Published: 2012 Citation: " Yan J, Jiang X, West AA, Perry CA, Malysheva OV, Devapatla S, Pressman E, Vermeylen F, Stabler SP, Allen RH, Caudill MA. Maternal choline intake modulates maternal and fetal biomarkers of choline metabolism in humans. Am J Clin Nutr. 2012;95:106071.


Progress 07/01/10 to 06/30/11

Outputs
OUTPUTS: Project 1: We assessed iron status, inflammatory markers and the iron regulatory hormone, hepcidin, in maternal and cord blood from a cohort of pregnant teens and are collecting data on relationships in women carrying multiples. Results were discussed in our undergrad course Human Anatomy and Physiology. Presentations and tours were provided to minority biology students. Analysis of data provided opportunities for undergrad and grad students to engage in our research. Several students are using study samples to undertake undergrad honors projects. Outreach included seminars at the University of Georgia and the International Trace Elements in Man and Animals meeting in China (TEMA 11). Two doctoral student's dissertations include using study samples. This research has strengthened our Human Metabolic Research Unit as iron status indicators and inflammatory markers are analyzed in house. Project 2: We completed recruitment and data collection from half of the subjects. Ultrasonographic endpoints for follicle wave kinetics were generated for most participants and serum is being analyzed for reproductive and metabolic hormones. We've made strides in developing/validating methods for the automated 3-dimensional (3D) assessment of sonographic images of normal and polycystic ovaries. These projects included participation from undergradstudents completing independent research studies. Preliminary findings of ovarian follicle growth kinetics in anovulatory women were presented at the annual symposia for the College of Agriculture and Life Sciences Research Honor's Thesis Program. Abstracts related to the establishment of refined criteria for the sonographic evaluation of ovarian morphology by 2D and 3D ultrasonography were accepted for presentation at the annual meetings of the Androgen Excess and Polycystic Ovary Syndrome Society and Canadian Fertility and Andrology Society. Project 3: We produced the first knockout mouse model designed to manipulate vitamin E, and possibly vitamin K, metabolism. Mice were bred, genotyped and placed on standard chow or soybean oil diets. We are evaluating how disruption of this gene, which encodes an enzyme with vitamin E catabolism activity (vitamin E-omega-hydroxylase), and which represents the only known vitamin E-metabolizing enzyme in mice, effects vitamin E status. We are using this mouse model to study the role of this enzyme in vitamin K metabolism and status, following up on observations in vitro that the analogous human enzyme (CYP4F2) also metabolizes vitamin K. We completed a study demonstrating that common genetic variants of this analogous human gene exhibit substrate-dependent alterations in the vitamin E-metabolizing activity of the enzyme. Project 4: In a controlled trial of choline supplementation, measurements of DNA strand breaks, plasma IL-6 and TNF-alpha in 20 pregnant and 20 nonpregnant women were performed, as well as micro-array analysis of placental tissue and circulating lymphocytes, and measurements of the epigenetic state of genes regulating fetal hypothalamic-pituitary-adrenal (HPA) activity and cortisol production. PARTICIPANTS: Kimberly O'Brien, PhD: Role: Co-investigator Partner Organizations: University of Rochester School of Medicine: Role: Provided access to patient population and regulatory infrastructure necessary to oversee human research regulations. Provided office space for health project coordinator who oversaw the implementation of the study. Collaborators and Contacts: Eva Pressman, MD - University of Rochester School of Medicine; Elizabeth Cooper, CNM, EdD - University of Rochester School of Medicine; Ronnie Guillet, MD, PhD - University of Rochester School of Medicine Training or professional development: Project 2: Marla Lujan, PhD, Role -Co-Investigator: Development and maintenance of Institutional Review Board (IRB) protocols; Participant recruitment and interviews; Data Collection (perform ultrasound scans, venipuncture, etc.); Manage processing biological samples, data analysis and daily operations of clinical research suite, computer imaging lab and wet lab; Train and mentor research personnel and students; Preparation of manuscripts and dissemination of findings. Anna Sear (Diagnostic Medical Sonographer); Participant recruitment and interviews; Data Collection (perform ultrasound scans, venipuncture, etc.); Process biological samples and participate in daily operations and maintenance of the clinical research suite. Magnolia Ariza (Research Associate); Process biological samples and participate in daily operations of the wet lab; Train and mentor research personnel and students; Analyze data, prepare of manuscripts and disseminate of findings. Project 3: Robert S. Parker - Co-Investigator Partner Organizations - N/A Collaborators and Contacts - D. Stec, Univ. Mississippi Medical Center: Provided viral variants of the human CYP4F2 gene. D. Manor, Case Western Univ.: Assisted with identification of the mouse ortholog of the CYP4F2 gene. Training or Professional Development: Sabrina Bardowell, PhD candidate. Conducted studies with CYP4F2 variants, and with the new mouse model described above. Project 4: Marie Caudill, Co-Investigator: Supervised the analytical measurements and participated in the data analysis, data interpretation and abstract development. TARGET AUDIENCES: Project 1: Adequate iron supply during pregnancy is critical for health and optimal development of the neonate. Insufficient iron transfer to the fetus across pregnancy can change the development trajectory of organs and pre-dispose the baby to chronic diseases later in life. Low iron stores at birth have also been linked to delayed infant motor and mental development. Few data at present exist on the impact of maternal anemia and maternal obesity during pregnancy on neonatal Fe stores or its potential to impact functional outcomes associated with Fe status. Moreover, little is known about Fe transport mechanisms in the placenta. Pregnant adolescents and women carrying multiples are key target groups to assess these relationships. Adolescents have additional competition for iron due to the maternal needs of growth coupled with fetal Fe demands and women carrying twins or triplets have substantially increased fetal iron demands and often have excessive gestational weight gains. The proposed studies target several research recommendations that were raised by the 2001 Institute of Medicine's Dietary Reference Intakes including; "Study of the effect of limited iron intake during pregnancy on infant iron status during the first 6 months of life" and "Concurrence of valid indicators for assessing the effect of iron deficiency anemia on cognitive development and function" and "integrative mechanisms of iron transporter proteins that influence absorption in various dietary conditions and physiological states". Results from these studies will have relevance to the larger population of pregnant women and neonates in the United States and will further our understanding of the functions and mechanisms of regulation of Fe during the reproductive period and the impact of maternal Fe status on neonatal Fe status at birth. Our findings from this racially diverse, pediatric and adult population provide novel data relevant to minority health and maternal /neonatal outcomes that can be used to better inform policy for public health benefit. Project 2: Women participating as research subjects receive valuable education on Female Reproductive Physiology, Nutrition and Metabolism and Reproductive Health. Participants include women from diverse racial and ethnic groups and females under the age of 21. This project has provided experiential learning opportunities for students in Nutrition, Engineering and Biological Sciences at Cornell through courses NS4010 Empirical Research and BIOG2990/4990 Independent Research. Project 3: Findings will be broadly applicable to human populations, shedding light on potential health outcomes of various forms of vitamin E that are rich in some types of vegetable oils. Certain findings may be of specific relevance to individuals carrying certain of the CYP4F2 genetic variants we have characterized. Project 4: Pregnant women and their offspring; Policy makers including the Food and Nutrition Board of Institute of Medicine; Health care providers; Research scientists working in the area of maternal and child health; Research scientists working in the area of one-carbon metabolism. PROJECT MODIFICATIONS: Project 1: No human subject concerns or major changes in the implementation of this research project have occurred. Project 2: The project has been significantly delayed because of the maternity leave of the PI who is directly involved in data collection from human subjects. The new construction and move of the Division's Clinical Chemistry Lab has delayed completion of assays for endocrine and metabolic markers. Project 3: We have deemphasized studies of a novel vitamin K metabolite, as a non-trivial amount of this metabolite occurs as an artifact of thermal breakdown of vitamin K epoxide in the gas chromatography-mass spectrometry method we employ. Emphasis was therefore shifted to the interaction between vitamin K and vitamin E, the metabolic basis of which remains a mystery. Project 4: The dramatic effects of maternal choline intake on placental gene expression motivated us to expand the project to include an investigation of the epigenetic state of the following genes: corticotrophin releasing hormone (CRH), glucocorticoid receptor (NR3C1), insulin-like growth factor 2 (IGF2), leptin (LEP), guanine nucleotide binding protein, alpha stimulating, antisense transcript (GNAS-AS1), and interleukin 10 (IL10). Selection of these genes was based on the gene expression results and on prior evidence of methylation regulation, susceptibility to nutritional exposures, and associations with chronic disease risk.

Impacts
Project 1: Studies to date have identified pregnant adolescents as particularly vulnerable group to iron deficiency. Our data indicate that the obesity often evident in this group is not inappropriately elevating hepcidin to limit Fe absorption. Our studies have also found that neonatal hepcidin is independently regulated which may speak to the ability of the fetus to control placental iron transport in response to fetal demands. The high prevalence of anemia found in these pregnant teens and the low iron stores often evident in neonates born to this group indicate that additional screening and targeted supplementation interventions are needed in this high risk group. Project 2: Preliminary data show clear fluctuations in ovarian follicle populations over time which challenges the notion that follicles in polycystic ovaries are "arrested" in development. Ultrasonographic criteria for normal and polycystic ovarian morphology have also been revised and proposed for use in clinical practice. Since longitudinal assessments of follicle growth kinetics are incredibly labor intensive and require a high degree of experience from the operator, we have implemented use of 3D ultrasound imaging. We have validated the use of automated follicle counting software for the longitudinal assessment of normal and polycystic ovaries. The application of this technology in our laboratory is critical for the timely completion of the proposed studies. Project 3: RT-PCT revealed that RNA expression of the targeted gene was completed absent in the knockout mice, and reduced by 50 percent in heterozygous mice. Knockout mice exhibited a 92 percent reduction in gamma-tocopherol catabolism capacity in the liver, but only 70 percent reduction in alpha-tocopherol catabolism capacity. This suggests the presence of an alternative enzyme, of unknown identify, possessing this activity. Vitamin E metabolites were substantially reduced, and accompanied by a hyper-accumulation of gamma-tocopherol in mice fed soybean oil. Knockout mice exhibited a more than proportional response to an increase in dietary vitamin E intake, in contrast to wild type mice. This model should prove useful in identifying novel biological activities of gamma tocopherol in vivo. Project 4: The higher maternal choline intake led to down-regulation (P<0.01) of genes involved in stress response, inflammation and preeclampsia. The higher maternal choline intake also yielded: (i) higher placental promoter methylation of the cortisol regulating genes, corticotropin releasing hormone (CRH; P = 0.06) and glucocorticoid receptor (NR3C1; P < 0.01); (ii) lower (P < 0.05) placental CRH transcript abundance; and (iii) lower (P ≤ 0.05) promoter methylation of both genes (CRH and NR3C1) in cord blood leukocytes. These data suggest for the first time in humans that varied maternal choline intake modulates the epigenomic status of the placenta as well as the epigenetic state of genes that regulate fetal HPA axis activity.

Publications

  • Cao C, Bemis T, Young MF, Essley BV, Pressman EK, McNanley T, Westerman M, OBrien KO., 2011. Placental Expression of Proton Coupled Folate Transporter/Heme Carrier Protein 1 is Related to Neonatal Hepcidin, Maternal Iron Status and Placental Expression of Heme Oxygenase in Pregnant Adolescents. In preparation
  • Bemis T, Foehr M, Jaacks L, McNanley TJ, Cooper EM, Pressman EK, McIntyre AW, Abkowitz JL, Guillet R, OBrien KO. 2011. Placental heme oxygenase-2 in pregnant adolescents and FLVCR expression. FASEB J;25:607.13.
  • Cao C, Young M, Kent T, Cooper E, Pressman E, McIntyre A, McNanley T, Westerman M, Guillet R, OBrien K., 2011. Serum haptoglobin: a marker of maternal obesity and neonatal iron status. FASEB J; 25:607.11
  • Lujan ME, Brooks ED, Pierson RA, Peppin AK, Chizen DR. 2011 Revisiting ultrasonographic criteria for polycystic ovarian morphology: new thresholds for elevated follicle population. Proceedings of the 93rd Annual Meeting of the Endocrine Society (Accepted).
  • Pawlak AC, Lujan ME. 2011. Longitudinal characterization of follicle populations in women with PCOS. Proceedings of Cornell University College of Agriculture and Life Sciences Research Honors Thesis Program Symposia. Ithaca, NY, United States.
  • Bardowell SA, Stec DE and Parker RS. 2010. Common variants of cytochrome P450 4F2 exhibit altered vitamin E omega-hydroxylase specific activity. J Nutrition 140:1901-1906.
  • Jiang X, Yan J , West AA, Perry CA, Malysheva OV, Bar H, Wells M, Devapatla S, Pressman E, Caudill MA. 2011. A higher maternal choline intake favorably alters placental gene expression of biological pathways related to disease risk. Experimental Biology, Washington DC.
  • Yan J , Jiang X, West AA, Perry CA, Malysheva OV, Devapatla S, Pressman E, Caudill MA. 2011. Maternal choline intake alters fetal indicators of stress-responsive hypothalamic-pituitary-adrenal (HPA) axis activity. Experimental Biology, Washington DC.
  • OBrien KO, Cao C, Young MF, McNanley TJ, Cooper EM, Pressman EK, McIntyre AW, Westerman M, Guillet R., 2011 Serum hepcidin at mid-gestation is not associated with neonatal hepcidin or maternal obesity among pregnant adolescents. FASEB J; 25:238.4.
  • Lujan ME, Jarrett BY, Pawlak AC. 2011 Longitudinal characterization of follicle populations in women with polycystic ovary syndrome by 2- and 3- dimensional ultrasonography. Proceedings of the Annual 9th Meeting of the Androgen Excess and PCOS Society (Accepted).
  • Lujan ME, Brooks ED, Pierson RA, Peppin AK, Chizen DR. 2011 Validated criteria for the ultrasonographic detection of polycystic ovarian morphology: new thresholds for elevated follicle population and ovarian volume. Proceedings of the Annual 9th Meeting of the Androgen Excess and PCOS Society (Accepted). Winner of the 2011 Azziz Baumgartner Travel Award For Young Investigators.
  • Jarrett BY, Lujan ME. 2011 Automated 3D-ultrasonography for the characterization of follicle populations in women with polycystic ovary syndrome. Proceedings of the 57th Annual Meeting of the Canadian Fertility & Andrology Society (Accepted).