Grantee Research Project Results
2012 Progress Report: Formative Center for the Evaluation of Environmental Impacts on Fetal Development
EPA Grant Number: R834594Center: Formative Center for the Evaluation of Environmental Impacts on Fetal Development
Center Director: Boekelheide, Kim
Title: Formative Center for the Evaluation of Environmental Impacts on Fetal Development
Investigators: Boekelheide, Kim , De Paepe, Monique E , Thanos, Christopher , Phipps, Maureen , Brown, Phil , Gruppuso, Phillip , Kostadinov, Stephan
Current Investigators: Boekelheide, Kim , De Paepe, Monique E , Thanos, Christopher , Phipps, Maureen , Brown, Phil , Gruppuso, Phillip
Institution: Brown University , Women & Infants Hospital of Rhode Island
EPA Project Officer: Hahn, Intaek
Project Period: February 15, 2010 through February 14, 2014
Project Period Covered by this Report: February 15, 2012 through February 14,2013
Project Amount: $1,189,575
RFA: Children's Environmental Health and Disease Prevention Research Centers: Formative Centers (with NIEHS) (2009) RFA Text | Recipients Lists
Research Category: Children's Health , Human Health
Objective:
RD834594C001: Liver and the Metabolic Syndrome
Study the effect of environmental toxicants on human fetal development and the fetal origins of metabolic dysfunction in the offspring.
RD834594C002: The Effects of Endocrine Disruption on the Developing Human Fetal Prostate
Project 2 has been guided by the following working hypothesis: Exposure of the human fetal prostate to estrogenic chemicals during development results in epigenetic modifications that alter differentiation and predispose to carcinogenesis. This hypothesis is being tested by performing the following Specific Aims:
Specific Aim 1. Characterize the differentiation of human fetal prostate xenotransplants using immunohistochemical markers and gene expression following a time-course study.
Specific Aim 2. Examine the differentiation and carcinogenesis of human fetal prostate xenotransplants following estrogenic exposures.
Specific Aim 3. Investigate the epigenetic alterations induced by estradiol, genistein and bisphenol A exposure during human fetal prostate development using a genome-wide approach.
RD834594C003: Lung, Arsenic Exposure and Tissue Remodeling
This project is guided by the hypothesis that arsenic exposure of human fetal lung xenotransplants results in disrupted lung growth mediated by molecular and epigenetic alterations. The Specific Aims of the original application have remained unchanged. They are to: (1) develop and characterize a human fetal lung xenograft model of in utero arsenic exposure; (2) determine the effects of arsenic exposure on growth dynamics and gene expression in the developing human lungs; and (3) determine arsenic-induced epigenetic changes in the developing human lung.
Progress Summary:
RD834594C001: Liver and the Metabolic Syndrome
Our research project is still focused on the same overall goal—to assess the effect of environmental factors on the fetus with an emphasis on fetal origins of metabolic syndrome in the offspring. During the first year of the proposal, it became apparent that fetal liver transplantations were not feasible because of hepatocyte loss. We therefore initiated studies on a white adipose tissue (WAT) xenograft model.
The WAT model has proved successful. Implants are viable for up to 4 months. They display the normal histological phenotype of adipose tissue at all the time points beyond 2 weeks, which is the earliest date at which the transplants become established. Immunohistochemical characterization of the transplants indicates ongoing adipogenesis that declines slightly with time but is nonetheless persistent. We have also observed very active angiogenesis in these transplants.
It is well established that prenatal factors predispose the offspring to development of obesity, metabolic syndrome and type 2 diabetes. The significance of this project lies in its potential to develop a model system to study the relevant pathophysiological mechanisms. This is a goal that is not possible with available methodologies. Achieving this goal would permit us to study the effect of environmental toxicants on fetal development relating to postnatal metabolic syndrome and obesity. In addition, we hope to succeed in establishing a well-characterized model that can be used by other investigators.
RD834594C002: The Effects of Endocrine Disruption on the Developing Human Fetal Prostate
The basic characterization of the human fetal prostate xenotransplant model has been completed (Specific Aim 1). Using a wide variety of immunohistochemical markers, we have shown the progressive differentiation and proliferation of the stromal and epithelial compartments after xenotransplantation. Examining the implants at 7, 30, 90 and 200 days post-transplantation, epithelial and stromal compartments demonstrate differentiation as indicated by the epithelial marker prostate specific antigen (PSA), as well as the stromal marker caldesmon. The differentiation markers detected by immunohistochemistry are associated with complementary DNA methylation epigenetic modifications. For example, the promoter regions of proteins highly expressed in the stroma are relatively hypomethylated compared to these same regions in epithelium, and vice versa. In summary, our characterization of this model demonstrates the utility of the xenotransplant approach for studying human prostate development and possible disruptions of development resulting from endocrine disrupting chemical exposure.
We have completed the collection of samples and will soon begin to determine the effects of estrogen exposure on human fetal prostate xenografted into both rat and athymic nude mouse hosts (Specific Aim 2). The dosing paradigm evaluates the effects of exposure to estradiol at both early (7 and 90 days) and later-life time points (200 and 250 days) of human implants following xenotransplantation. After implantation, xenografts receive an “early-life” exposure to either corn oil or estradiol every other day for a total of three injections. Some xenografts are collected at 7 and 90 days, while those receiving a “later-life” exposure receive an additional dose of estrogen beginning at day 90 for 110 days (until 200 days postimplantation). Some xenografts are immediately collected at day 200, while some are allowed a recovery period and are collected at day 250 (Figure 1).
Figure 1.
Additionally, the endogenous prostates of the rodent hosts have been examined to address the species effects of hormonal manipulations. The results to date show estrogen-induced basal cell hyperplasia of the human fetal prostate transplants, a response that is modest in comparison to the exuberant hyperplasia of the endogenous rodent prostates.
Currently underway is a detailed immunohistochemical and molecular analysis of these hyperplastic responses.
RD834594C003: Lung, Arsenic Exposure and Tissue Remodeling
Our work has focused on continued optimization and validation of the human fetal lung xenograft model. In addition, studies were initiated to determine the effects of arsenic exposure on growth dynamics and gene expression in the developing human lung. Our results indicated that exposure of human lung xenografts to 100 ppb arsenic results in morphologic alterations (including to the microvasculature) as well as differential gene expression, including upregulated expression of several angiogenesis-regulating genes (ANGPT2 and ESM1). Interestingly, arsenic exposure has been implicated in altered angiogenesis in several other organ systems. It is tempting to speculate that the apparent effects of arsenic exposure on lung development may be mediated, in part, by its effects on microvascular remodeling. Initial microarray analyses have shown trends towards altered expression of genes involved in regulation of extracellular matrix remodeling, angiogenesis and immune regulation, but need to be confirmed in larger studies.
These studies will give insight into the mechanisms of arsenic-induced respiratory disease and may lead to the elucidation of molecular targets and biomarkers of inorganic arsenic exposure during lung development. Furthermore, this study provides a framework for future studies investigating other environmental toxicants.
Community Outreach and Ttranslation Core Project Summary/Accomplishments
The Community Outreach and Translation Core (COTC) served three important functions for the Center: (1) established partnerships with health care, government and community organizations; (2) facilitated educational and outreach opportunities to build capacity among faculty, health care practitioners and Rhode Islanders; and (3) investigated ethical issues regarding informed consent in fetal tissue research as well as health care practitioners’ knowledge of the (fetal) developmental origins of health and disease (DOHaD).
The COTC employed a dynamic, multitiered community engagement strategy to target key audiences. The COTC formed two active coalitions: the Community Advisory Board (CAB) to engage Rhode Island organizations and agencies, and Hospitals for a Healthy Environment in RI (H2ERI) to engage the health care community. Additionally, COTC staff served as active board members for a number of coalitions and organizations that address children’s health issues. The COTC has organized successful conferences, workshops and presentations on environmental health topics and encouraged environmentally sustainable practices to reduce environmental exposures detrimental to children’s health. In addition to outreach, COTC has played a key role in setting rigorous ethical standards for fetal tissue research and interacting with study participants.
The COTC has been shaped by its highly involved CAB and community-based partners. The CAB met monthly during the planning phase of the Center’s formation and then quarterly to guide this Core. The CAB is comprised of 10 members from Rhode Island community-based organizations, disease-based charities, state agencies and education who are actively involved in children’s health and welfare: Environmental Justice League of Rhode Island, Childhood Lead Action Project, Meeting Street, Ready to Learn, Rhode Island Department of Health (HEALTH), March of Dimes, American Lung Association, Lincoln School and Project HEALTH. CAB members were invited to the Center’s monthly seminars and its annual retreat. They received notifications of all Center-related activities. In turn, some CAB members presented at the Center’s annual retreat and epigenetics conferences. The CAB provided one avenue by which the COTC assessed the effectiveness of their developed products.
The COTC was involved in pharmaceutical waste reduction and medical waste management. The COTC joined the Rhode Island Department of Environmental Management’s (RIDEM) task force, developed related undergraduate student projects in nursing and conducted master’s thesis research in environmental studies. These activities have provided groundwork for new RIDEM-related policies and practices. Additionally, Dr. Thompson (Health Specialist) briefed members of the Rhode Island State Senate on these issues at their request.
The COTC formed the H2ERI Coalition. This Coalition began as a COTC partnership with nurses at Women and Infants Hospital (WIH) to remove DEHP- and PVC-based medical equipment from its Neonatal Intensive Care Unit (NICU). The Coalition’s formation was deeply influenced by the expressed needs and suggestions of Rhode Island-based hospital managers, health care professionals, union leaders, and members of food policy groups and community-based health organizations. Under the guidance of the CAB with assistance from Health Care Without Harm, Practice Greenhealth and Maryland Hospitals for a Healthy Environment, efforts were expanded to encompass a comprehensive approach to environmental sustainability in health care settings. This initiative was widely successful. Two annual conferences on best practices for environmental excellence in health care have been held (80 and 100 attendees, respectively). Ten Brown students were recruited to help organize both conferences as a way to educate them and to spread word of the Center’s work throughout the Brown campus. Today, the coalition has 50 members from 31 different organizations representing hospitals, hospital associations, state and federal agencies, and nongovernmental organizations (NGOs). The Coalition meets bimonthly to plan activities and showcase successful programs and initiatives. COTC postdoctoral fellow Dr. Panikkar represented the H2ERI coalition at the national CleanMed conference and helped Miriam Hospital in Providence appoint an environmental sustainability coordinator. Dr. Panikkar gave presentations on topics including DEHP/PVC in medical devices, green team building and environmental sustainability practices in health care. At the 2012 annual CEHC/Partnerships in Environmental Public Health meeting, Dr. Brown led a workshop to help other Centers build similar healthy hospital coalitions.
The COTC was actively involved in community partnerships. The COTC participated in environmental health policy and public health campaign initiatives through the Environmental Justice League of Rhode Island and Childhood Lead Action Project. Additionally, Core staff became members of Alliance for a Healthier Rhode Island (AHRI); LAUNCH, a federal mental health and substance abuse program; and the HUD-funded Green and Healthy Housing Initiative. The COTC was very involved with AHRI’s campaign to impose an excise tax on sugar-sweetened beverages and curtail distribution in schools, hospitals and public facilities. Dr. Panikkar and other AHRI members briefed Rhode Island Attorney General Peter Kilmartin on this issue. Additionally, Dr. Panikkar established a healthy beverage work group. COTC members advised RIDOH’s Maternal, Infant and Early Childhood Home Visiting Program and the Local Wellness Council (LWC), an organization that addresses the needs of developmentally disabled children ages 0 to 8. Dr. Thompson served as an advisor to Rhode Island’s Strategic Plan for Preconception Health to address preconception health through clinical practice, health policy and public health initiatives. The COTC cosponsored the state’s Preconception Health Summit.
The COTC held a workshop on Epigenetics and Fetal Origins of Health and Disease, a 2-hour workshop held for 83 health care professionals, faculty and students from Rhode Island and Massachusetts. Through the efforts of Dr. Thompson and the Rhode Island State Nurses Association, registered nurse (RN) attendees received continuing education credit not only for this workshop but the second H2ERI conference as well.
The COTC provided Environmental Health Training for Home Visitors. Drs. Brown and Panikkar, and CAB members, developed a training program and conducted two statewide workshops on children’s environmental health for Meeting Street early intervention staff and Bradley Research Center home visitors who work with developmentally disabled children and their families. The workshop highlighted how children are uniquely susceptible to environmental hazards during their earliest years. The workshop illustrated ways to avoid or reduce residential exposures.
The COTC organized Science Cafés. The COTC organized a series of Science Cafés in local coffee shops and restaurants on relevant environmental health topics in conjunction with the Silent Spring Institute.
The COTC cosponsored Community Environmental College and ECO-Youth. These summer and after school train-the-trainer programs allowed 65 high school students to teach others about environmental health issues, including asthma, obesity and reproductive health. Since 2009, the COTC has cosponsored these programs with Brown’s Superfund Research Program and the Environmental Justice League of Rhode Island.
The COTC conducted ethics research. Ethics is an integral part of the Center’s work. The Center’s team acknowledged that fetal tissue xenotransplantation research could be a publicly contentious issue. To ensure that ethical guidelines would be well integrated with the Center’s research projects, Dr. Brown held a series of meetings with the former Health Specialist and the Tissue Procurement Core Leader to discuss specific project-related ethical issues. These meetings shaped the ethical issues that the Center addresses. Many ethical issues arose in designing these research projects. Dr. Brown led the Center’s team in evaluating the informed consent process through participant interviews. This preliminary work resulted in a manuscript that contributed to the literature on bioethics and advanced forms of informed consent (Panikkar et al., 2012).
The COTC assisted with research on evidence-based practices. Dr. Gruppuso (the Center’s former Health Specialist) and Dr. Brown initiated a project to assess physicians’ knowledge of DOHaD. Alisha Lakhani, M.P.H., conducted structured interviews with six OB/GYN, family practice and pediatric physicians. These interviews revealed several unexpected findings. The physicians questioned the validity of DOHaD. They were hesitant to incorporate DOHaD-related elements into their current practice of counseling prenatal clients. Also, they were concerned about generating parental guilt and stress. However, some of these physicians identified environmental toxins as the “next thalidomide.”
Future Activities:
RD834594C001: Liver and the Metabolic Syndrome
We will carry out xenotransplantation of peri-renal adipose tissue from human fetuses and adult humans to immunodeficient mice. The goal will be to compare rates of angiogenesis and adipogenesis in fetal and adult tissues. We will be testing the hypothesis that fetal WAT is better able to sustain rapid expansion without pathologic changes owing to higher rates of angiogenesis.
We are also going to analyze pre-transplant fetal liver and WAT for DNA methylation status. Comparisons will be made to normal adult liver and WAT.
RD834594C002: The Effects of Endocrine Disruption on the Developing Human Fetal Prostate
The ongoing experiments with estrogenic exposure of the human fetal prostate model are reaching completion and are undergoing detailed analysis at this time. Based on the data obtained to date, this is a very useful model for examining endocrine disrupting chemical effects on the developing human prostate.
RD834594C003: Lung, Arsenic Exposure and Tissue Remodeling
We will complete the study of the effects of arsenic exposure on lung growth dynamics (morphometric and proliferation analyses) and gene expression (additional microarrays, additional confirmatory analyses). In addition, we will start to study of the effects of arsenic exposure on lung epigenetic marks using the material assembled during the previous years.
Journal Articles: 8 Displayed | Download in RIS Format
Other center views: | All 13 publications | 8 publications in selected types | All 8 journal articles |
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Boekelheide K, Blumberg B, Chapin RE, Cote I, Graziano JH, Janesick A, Lane R, Lillycrop K, Myatt L, States JC, Thayer KA, Waalkes MP, Rogers JM. Predicting later-life outcomes of early-life exposures. Environmental Health Perspectives 2012;120(10):1353-1361. |
R834594 (Final) R834594C002 (Final) |
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De Paepe ME, Chu S, Hall S, Heger NE, Thanos C, Mao Q. The human fetal lung xenograft: validation as model of microvascular remodeling in the postglandular lung. Pediatric Pulmonology 2012;47(12):1192-1203. |
R834594 (2012) R834594 (Final) R834594C003 (Final) |
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De Paepe ME, Chu S, Heger N, Hall S, Mao Q. Resilience of the human fetal lung following stillbirth: potential relevance for pulmonary regenerative medicine. Experimental Lung Research 2012;38(1):43-54. |
R834594 (2012) R834594 (Final) R834594C003 (Final) |
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Heger NE, Hall SJ, Sandrof MA, McDonnell EV, Hensley JB, McDowell EN, Martin KA, Gaido KW, Johnson KJ, Boekelheide K. Human fetal testis xenografts are resistant to phthalate-induced endocrine disruption. Environmental Health Perspectives 2012;120(8):1137-1143. |
R834594 (Final) R834594C002 (Final) |
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Panikkar B, Smith N, Brown P. Reflexive research ethics in fetal tissue xenotransplantation research. Accountability in Research 2012;19(6):344-369. |
R834594 (2012) R834594 (Final) |
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Saffarini CM, McDonnell EV, Amin A, Spade DJ, Huse SM, Kostadinov S, Hall SJ, Boekelheide K. Maturation of the developing human fetal prostate in a rodent xenograft model. The Prostate 2013;73(16):1761-1775. |
R834594 (2012) R834594 (Final) R834594C002 (Final) |
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Spade DJ, Hall SJ, Saffarini CM, Huse SM, McDonnell EV, Boekelheide K. Differential response to abiraterone acetate and di-n-butyl phthalate in an androgen-sensitive human fetal testis xenograft bioassay. Toxicological Sciences 2014;138(1):148-160. |
R834594 (Final) R834594C002 (Final) |
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Thompson MR, Boekelheide K. Multiple environmental chemical exposures to lead, mercury and polychlorinated biphenyls among childbearing-aged women (NHANES 1999-2004): body burden and risk factors. Environmental Research 2013;121:23-30. |
R834594 (Final) |
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Supplemental Keywords:
Adipogenesis, in utero exposure, mammalian, metals, developmental biology, perinatal programming, human fetal prostate, differentiation, endocrine disrupting chemicals, arsenic, children's environmental health;, Health, Scientific Discipline, ENVIRONMENTAL MANAGEMENT, Health Risk Assessment, Risk Assessments, Biology, Risk Assessment, biological pathways, children's health, fetal exposure, bioavailability, developmental effects, perinatal exposure, biochemical researchProgress and Final Reports:
Original Abstract Subprojects under this Center: (EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
R834594C001 Liver and the Metabolic Syndrome
R834594C002 Prostate and Endocrine Disruption
R834594C003 Lung, Arsenic Exposure, and Tissue Remodeling
The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.