I am an environmental epidemiologist with a primary research interest in the role of developmental windows, joint impact of multiple exposures and molecular mechanisms through which prenatal/early-life exposure to chemical and non-chemical stressors influence children's lung development. The goal of this proposal is to obtain training in the skills needed to continue to build my academic career by linking ambient air pollution and stress to lung development and to identify biomarkers of early biological effects of these exposures. Formal coursework and the expert mentorship of Drs. Rosalind Wright, Andrea Baccarelli, Robert Wright, Maria Martha Tellez-Rojo, Brent Coull, Alfin Vicencio and Allan Just will enable me to acquire the knowledge and skills necessary to become an independent transdisciplinary researcher. This training will allow me to achieve my long-term career goals: taking the lead in studying respiratory health in the ongoing Mexico City PROGRESS cohort in order to examine these and other environmental exposures, their biological mechanisms, and the subsequent effects on lung function and asthma risk longitudinally as these children grow up. All relevant exposure and covariate data exist in an existing well-phenotyped pregnancy cohort [Programming Research in Obesity, Growth, Environment and Social Stressors (PROGRESS) study leveraged herein. I will add respiratory phenotyping to the cohort. Specifically I will 1) train in state of the art stress assessment as well as fetal and early childhood lung development and physiology with Dr. RJ Wright, 2) train in early biologic markers of oxidative stress in Dr. Baccarelli's lab, 3) train in advanced statistical methods that temporal resolved daily to weekly assessed exposure data to definitively identify susceptibility windows to enhance our ability to find effects and identify vulnerable groups with Dr. Brent Coull, 4) integrate research on stress, air pollution and molecular epidemiology through interactions with my mentorship team to become an interdisciplinary scientist and 5) translate the research and administrative training I receive to establish myself as a independent investigator with a tenure track faculty position and to position myself for a future R01. This proposal will address gaps in the understanding of timing and mechanism through which air pollution exposure and stress affect respiratory health in early childhood. This proposed research and training plan builds the foundation for an independent research career that aims to clarify the mechanisms through which ambient air pollution and stress are associated with respiratory morbidity and ultimately inform intervention/prevention efforts. This study is also highly cost effective as we leverage the resources of the PROGRESS study with existing biospecimens, environmental and psychosocial stress data.

Mount Sinai will partner with the National Institute of Public Health, Mexico and Columbia University to continue to follow 700 children at ages 8-11 in the PROGRESS birth cohort established in 2006 in Mexico City to study perinatal programming of child development. PROGRESS is a state of the art birth cohort that has developed innovative new methods to estimate past exposure to environmental chemicals, both with biomarkers of internal dose (e.g. teeth based chemical assays) and with geospatial modeling of the external environment via remote sensing(e.g. air pollution, green space etc). This application will also leverage Mount Sinai's recently established Environmental Health Sciences Laboratory to build upon our internationally recognized research expertise in children's environmental health. Our mission is to measure environmental exposures across pregnancy and childhood to determine how these factors affect neurodevelopment and growth. We are especially excited to extend this work in PROGRESS as our overarching goal is to prepare the cohort the coming era of exposomic research. In this R24 we will test methods for data harmonization that will be critical to future work. By piloting pooling methods with researchers with similarly designed cohorts we set the stage to build new consortia to study the health impacts of chemicals, nutrition, social factors, and the gene-environment interactions that underlie childhood disease. Our team has all the requisite expertise in exposure science, biostatistics, child health phenotyping and in cohort management. We have already developed several new methodologies to objectively reconstruct past chemical exposures that precisely estimate exposure timing, allowing researchers to identify susceptibility windows as they relate to child health. Over the last 10 years, PROGRESS has supported 16 NIH grants led by 10 different researchers (including 7 K awardees), demonstrating our ability to serve as an incubator for the next generation of Environmental Health scientists. To accommodate divergent research needs among this group, the PI has formed the equivalent of an Administrative Core that will plan, organize, streamline and prioritize field work by coordinating the associated administrative work (IRB, protocol development, finances etc). We will also develop new programs to assess participant needs in order to improve retention, adapt to advances in social media, assess new health outcomes and exposures, and implement new exposure/response assays as we prepare our cohort for future big data consortia research. This proposal links highly experienced environmental health scientists with statisticians, social epidemiologists, child psychologists, chemists, pediatricians, and epigeneticists to support the infrastructure of a highly functional, state of the longitudinal birth cohort that objectively measures the human environments that program child health. We will follow PROGRESS children up to the onset of puberty, linking fetal exposures to health changes inherent to this critical life stage. Finally, despite our many successes, without this R24, field work will cease and this application is critical to our future success.

The overarching goal of the proposed research is to investigate the association of prenatal exposure to EDCs: phthalates and BPA) with DNA methylation patterns and childhood obesity using novel epigenomic approaches. The program provides mentored training (K99) and a research phase (R00) for Allan Just PhD, an environmental epidemiologist, to become an independent academic investigator in epigenetic epidemiology. Dr. Just will receive training and guidance from a mentoring team of internationally recognized scientists with specializations in environmental and epigenetic epidemiology, pediatrics, and biostatistics, including Dr. Andrea Baccarelli, Dr. Robert Wright and Dr. Xihong Lin. Prior research has demonstrated that human prenatal exposures can alter the fetal epigenome, and animal models have shown that phthalates and BPA impact methylation and obesity risk in the offspring. The proposed research uses methylation microarrays and new sequencing-based technologies to investigate methylation patterns in the umbilical cord blood of two cohorts of human children. An epigenome-wide methylation microarray (Illumina Human Methylation 450K BeadChip), already being conducted on umbilical cord blood DNA from 120 children, will measure methylation in 480K individual sites spanning almost all known genes. Statistical models will relate these methylation measures to prenatal EDC exposures and to measures of obesity at age 4. These models use new statistical approaches that account for multiple comparisons and reflect the dependence between methylation sites within the genome. Replication in an independent cohort (n=150), using the same design, assures that findings are consistent and generalizable. All samples and obesity measures are already collected or ongoing in the two cohorts (called PROGRESS and PRISM) under existing grants led by members of the mentoring team. In the R00 phase, a novel technique, targeted enrichment, which has not yet been applied in large-scale epigenetic epidemiologic studies, will be used in conjunction with bisulfite treatment and next generation sequencing; this approach will generate new high resolution measures of all methylation sites in identified regions of interest. This research should provide greater power to detect associations between EDC exposures, methylation patterns, and obesity in children. This approach may also contribute to future epigenetic epidemiological studies by comparing the contributions of the sparser, genome- wide platform with patterns found by the novel high density sequencing-based measures. This research is important because it quantifies the extent to which prenatal exposures to EDCs are associated with methylation patterns in human fetal tissue, and then links these findings to childhood obesity. Another innovation is its techniques to conduct epidemiologic studies with two different methylation technologies. The proposed work trains Dr. Just in state-of-the-art epigenomic approaches and in this way contributes to understanding of the impact of endocrine disruptors on children's health.

Obesity and related cardio-metabolic health disorders are leading causes of morbidity and mortality among women. The rapidly increasing worldwide prevalence of these disorders suggests that risk factors other than genetics, diet, and physical activity are involved. Environmental factors, such as endocrine disrupting chemicals (EDC), likely play an important role in modifying the response to dietary nutrients. Bisphenol-A (BPA) and phthalates are examples of EDCs that are ubiquitous in our environment and may contribute to the development of metabolic disorders by interfering with glucose and lipid metabolism and energy balance. Much of the current research focuses on exposures during stages of early life growth and development. However, pregnancy is a relatively unappreciated sensitive life stage in adults that is associated with numerous physiologic changes, which may have lasting consequences for women's health. Associations of exposures to BPA and phthalates during pregnancy and the development of cardio-metabolic health indicators in women during the long-term postpartum period have not yet been investigated. The candidate has a strong foundation in the methodological and analytical skills related to nutritional and reproductive epidemiology. The intent of this application is in seeking a Pathway to Independence Award to gain the additional training needed to accomplish the following long-term career goal: to establish an independent research program to investigate the intersection of environmental and nutritional factors related to disease development, progression, and treatment. The training component of this project will be based in the Department of Preventive Medicine (DPM) at Mount Sinai School of Medicine. The DPM is internationally renowned for excellence in innovative, interdisciplinary research in public health, epidemiology, environmental health, and social sciences. The mentors, Drs. Susan Teitelbaum and Robert Wright, are experts in the field of environmental health and the advisory team, Drs. Mary Wolff, Andrea Baccarelli, Brent Coull, and Mara Tellez-Rojo, possesses complementary areas of expertise to address training and research goals. The proposed training plan includes formal coursework, directed readings, attendance to scholarly seminars and meetings, and mentored career development activities, as well as rotations in Dr. Wright's molecular biology and analytical chemistry laboratories. From this training, the idea is to gain knowledge and skills related to environmental health and EDCs, toxicology, epigenetics, and advanced statistical analysis, which will be applied during the research component of this project. The proposed research will determine whether exposures to BPA and phthalates during pregnancy and the postpartum are associated with the development of adiposity and cardio-metabolic health indicators in women. This research will be conducted by leveraging data regarding maternal exposures and health outcomes collected from the Programming Research in Obesity, Growth, Environment, and Social Stressors (PROGRESS) study (R01 ES013744 and R01 ES021357 BPA, Phthalates & Stress: Mechanisms and Interactions for Childhood Obesity), a prospective birth cohort in Mexico. The following aims will be accomplished: 1) Determine whether exposures to BPA and phthalates in pregnancy and the postpartum are associated with the development of adiposity and cardio-metabolic health indicators in adult women up to 7 years postpartum. 2) Determine whether exposure to BPA and phthalates modifies the association of dietary fat with the development of adiposity and cardio-metabolic health indicators. 3) Determine the role of DNA methylation as a mediator of the associations of BPA and phthalates and the development of adiposity and cardio-metabolic health indicators. DNA methylation of nuclear receptor genes that are involved in body weight homeostasis will be measured: glucocorticoid receptor, ERα, and PPARγ. The training and research activities will result in scientific presentations and publications, and will prepare me to successfully compete for R01 funding during the R00 phase.

Childhood obesity is a growing crisis in the U.S. and worldwide. While increased caloric intake plays an obvious role, the underlying causes of this epidemic are likely not as simple as just caloric imbalance. The increase in obesity rates has been accompanied by extensive changes in our society. Animal models indicate that two features of these changes, i.e., increased manufacturing and exposure to endocrine disrupting chemicals (EDC) and the ever increasing burden from psychosocial stress, may be linked to obesity. In particular, both EDCs and stress can program obesity risks starting as early as in fetal life. In animals, fetal exposure to environmental endocrine disrupting chemicals (EDC), such as bisphenol A (BPA) and phthalates, reprograms the individual metabolic set point toward obesity via altered hypothalamic-pituitary-adrenal (HPA) signaling. The HPA axis is a central system for metabolic programming and a primary mediator of stress responses through cortisol production. In animals, fetal exposure to increased maternal stress causes HPA reprogramming, cortisol dysregulation, and higher risk of obesity. Not only do stress and EDCs regulate cortisol metabolism, both stress and EDCs have been shown to alter DNA methylation, a potential mechanism of HPA axis programming. DNA methylation regulates the glucocorticoid receptor and is sensitive to the environment, suggesting that epigenetics may underlie links between obesity, stress and EDCs. To our knowledge, interactions of psychosocial stress with BPA/phthalates have never been studied in humans, nor have human studies assessed cortisol metabolism or epigenetic biomarkers in research on EDCs and obesity. We will leverage the infrastructure and resources of the PROGRESS study (Programming Research in Environment and Social Stressors) in Mexico, a prospective birth cohort focused on early childhood neurocognitive phenotypes. PROGRESS provides a unique opportunity to perform research on obesity and metabolic dysfunction at economy of scale. In this proposal, we will add to PROGRESS by: 1) phenotyping children for obesity and metabolic dysfunction; 2) measuring EDC chemical exposure from archived urine; and 3) examining DNA methylation measures from archived cord blood. We will determine whether prenatal exposure to BPA & phthalates predicts altered trajectories of BMI from birth till 7 years, as well as greater adiposity, estimated insulin resistance, blood pressure, adipokines, and fasting lipid profiles at ages 4 & 7 years. We will then test whether BPA and phthalate exposures disrupt the maternal-fetal HPA axis, as indexed by salivary cortisol rhythms. We will examine whether the effects of BPA and phthalates on the obesity-related phenotypes and HPA axis are worsened in the presence of conditions of maternal psychosocial stress during pregnancy. Finally, we will explore the role of DNA methylation as a molecular mechanisms contributing to programming of HPA responses as a factor contributing to childhood obesity and metabolic dysfunction.