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Integration of Biological Cortisol Concentrations and Stress Questionnaires As Markers of Maternal Stress Doi:10.1016/J.Ntt.2014.04.012 Platform presentations 79 data. Our aims were to integrate high dimensional placental multi'- placenta at delivery. A process for collecting, storing, shipping, and omics data sets with measured environmental contaminants and analyzing placentae from 11 US counties in CA, MN, NC, NY, PA, pregnancy outcome data. Study Design: As part of the NCS Main Study, SD, UT, and WI was developed, standardized, validated, and im- 210 placentas from 10 regional centers were uniformly collected and plemented for assessment of morphology/pathology, genetics, sys- stored with a priori protocols designed to preserve biologic material tems biology, and environmental exposures in the same placentae. for future 'omics research. Subsequent DNA/RNA/miRNA/methylated All tissues were sent to a central placental processing center (UR). DNA analysis, analytic chemistry for environmental exposures, and 3-D Methods: Placentae (N = 253) were studied. Following delivery, placental imaging and mapping were conducted by established experts placental tissue was frozen and stored at -80 °C. Mass Spectrometry from over 20 collaborating centers. High dimensional data sets were (MS) was used for all environmental assessments: 45 metals (ICP- reduced and integrative analysis to determine the interrelationships MS); Bisphenol A (BPA) (LC-MS(quadrapole)); and 10 Polybro- among biologic measures was undertaken. RNA levels for all annotated minated diphenyl ethers (PBDEs)/ 32 polychlorinated biphenyls(PCBs)/ transcripts (UCSC & Ensembl) were normalized to construct a weighted dichlorodiphenyldichloroethylene (DDE) (GC–MS/MS). Results: Asub- coexpression gene network. Leveraged variant information derived study of 43 placentae refrigerated over 96 hrs demonstrated that all from placental RNA sequencing enabled construction of a weighted environmental analyses were stable over at least 72 h following delivery. gene network of 20,000 most variable gene expression traits. 1 million For all 253 placentae, geographical differences in placenta PBDE congener SNPs and SNPs from 1000 genomes (14 million SNP genotypes per concentrations were noted but not for BPA, PCBs or DDE. Interestingly, sample) were imputed across all samples, and placental eQTL were approximately six percent of all placentae had levels of gadolinium + 3 mapped to identify enriched loci at p b 0.00001. Results: A heretofore (Gd) that were 800–1000× higher than the other placentae. These unparalleled data set was generated and analyzed to yield novel elevated Gd levels were distributed throughout all geographical regions. placental interaction networks (Figure). The placental co-expression Discussion: These investigations demonstrated that placentae can be network indicates strong structurally segregated regions of the collected throughout the US and analyzed centrally under strict and placental molecular interactome, generating 20 modules, each con- controlled analytical conditions for monitoring environmental agents. taining between 30 and 4000 gene members. In testing for pathway Using uniform and validated methods for collection, storage and analyses, enrichments across all modules, we uncovered not only thousands of these studies demonstrated that for some environmental agents that loci modulating the expression of genes in the placenta, but one regional differences can be monitored, while other contaminants dem- subnetwork (yellow nodes/red edges) that was enriched for pathways, onstrate that subsets of the population can have elevated levels of the SNPs, and toxins previously associated with autism and associated selected contaminants (Gd) or have uniformly, very low levels (BPA). spectrum disorders. Conclusion: We demonstrate for the first time that (Supported in part by NIH LOI 2-BIO-18.) population-derived high dimensional placental multi'omics data can be integrated with systems biology approaches. These comprehensive doi:10.1016/j.ntt.2014.04.013 molecular profiles, when taken in the context of other high-dimensional phenotypic data, promise to enhance our understanding of a variety of biological processes that are modulated in the placenta and in turn associate with later human disease. NBTS 11 Integration of biological cortisol concentrations and stress questionnaires as markers of maternal stress doi:10.1016/j.ntt.2014.04.012 M.N. Smitha,B.Thompsonb,S.A.Beresforda,b,E.Vigorena,E.M.Faustmana aUniversity of Washington, Seattle, WA, United States NBTS 10 bFred Hutchinson Cancer Research Center, Seattle, WA, United States Environmental exposures during pregnancy: US National Children's Study formative research investigation High levels of maternal stress during the critical prenatal window of susceptibility are associated with lifelong adverse health outcomes R.K. Millera,b,T.H.Darraha,c,A.Friedmana,b,A.Lia,d,C.J.Stodgella,b, for the fetus. Consequences of maternal stress may include dis- P. Landrigana,e,C.Walkera,g,E.B.Clarka,i,N.Dolea,k,N.Thiexa,j,J.Swansona, ruption of the hypothalamic–pituitary–adrenal axis and increased h,J.Culhanea,l,S.Szaboa,f,J.Moyea,m risk of obesity and mental health disorders later in life. In order aNational Children's Study Placental Consortium, Bethesda, MD, United to better characterize these impacts it is necessary to understand States environmental factors impacting maternal stress. In this study we bUniversity of Rochester, Rochester, NY, United States demonstrate how biological measurement of cortisol (blood, saliva, cOhio State University, Columbus, OH, United States urine, and hair) can be integrated related to stress questionnaires dUniversity of Illinois at Chicago, Chicago, IL, United States to quantify both acute and chronic stress. Cortisol is released into eIcahn School of Medicine, New York City, NY, United States the blood in a pulsatile manner and quickly metabolized; therefore fMedical College of Wisconsin, Milwaukee, WI, United States instantaneous measurements in blood and saliva may not reflect gUniversity of California at Davis, Davis, CA, United States chronic stress. Urine cortisol concentrations reflect stress over recent hUniversity of California at Irvine, Irvine, CA, United States hours. Hair provides a retrospective calendar of cortisol concentra- iUniversity of Utah, Salt Lake City, UT, United States tions for up to six months. We have created a biokinetic model that jSouth Dakota State University, Brookings, SD, United States allows for these samples to be integrated across biological compart- kUniversity of North Carolina, Chapel Hill, NC, United States ments. This, in conjunction with saliva, blood, and urinary cortisol lThe Children's Hospital of Philadelphia, Philadelphia, PA, United States concentrations collected on consecutive days and across seasons mNational Institutes of Health, Bethesda, MD, United States allows for the characterization of both acute and chronic stress. Chronic stress is also commonly assessed by stress questionnaires. Introduction: Environmental exposures can potentially result in We have compared our saliva cortisol concentrations to two different toxicity, while in other instances; low levels of exposure may not stress questionnaire responses in two populations of women. In both produce a toxic impact. How can we monitor a human population the rural and urban cohorts, aspects of the stress questionnaires were for environmental exposures of concern? We investigate here the related to salivary cortisol concentrations. Neighborhood safety and feasibility of monitoring environmental toxicants in the human satisfaction were correlated with morning cortisol concentrations in the 80 Platform presentations rural cohort. In both cohorts the Mexican Farmworker questionnaire Developmental exposure to methylmercury (MeHg) may be was related to the Perceived Stress Scale. This study builds tools for unmasked later in life. In animal models, signs of poisoning may the integration and optimization of maternal stress assessment. Under- not appear for weeks or months after exposure, and in humans the standing the relationship between and within biological and ques- effects may be masked for months or years, perhaps even decades. tionnaire based stress measurements is important to the cohesive Mechanisms that govern the protracted unmasking of MeHg neuro- characterization of maternal stress exposure. This work was supported toxicity have yet to be fully uncovered. Potential mechanisms include the National Institute of Child Health and Human Development, (1) the slow production and accumulation of a toxic metabolite/s, National Institutes of Health, Department of Health and Human Services (2) compounding of the normal age-related cell loss by develop- (Contract No. HHSN267200700023C), by National Institute of Environ- mental MeHg exposure, (3) exhaustion of inherent compensatory mental Health Sciences (Award #5P01ES009601), by National Institute mechanisms, and (4) later-life exposure to Hg or other compounds of Diabetes and Digestive and Kidney Diseases (Award # R01 DK079042) may unmask the developmental effects of MeHg. The talk will discuss and by USEPA (grant #RD8345140). molecular mechanisms and genetic interactions associated with the unmasking of silent MeHg neurotoxicity drawing parallels from other diseases, which are known to affect the central nervous system, such doi:10.1016/j.ntt.2014.04.014 as Parkinson disease. doi:10.1016/j.ntt.2014.04.016 NBTS 12 Systems medicine and proactive P4 medicine: A revolution in healthcare NBTS 14 L. Hood Prenatal stress unmasks neurotoxic consequences of developmental Institute
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