Grantee Research Project Results
2011 Progress Report: Children's Environmental Health and Disease Prevention Research Centers: Formative Centers
EPA Grant Number: R834678Center: Center for Integrative Research on Childhood Leukemia and the Environment - 2015
Center Director: Metayer, Catherine
Title: Children's Environmental Health and Disease Prevention Research Centers: Formative Centers
Investigators: Woodruff, Tracey J. , Fisher, Susan J. , Steinauer, Jody , McMaster, Michael , Stotland, Naomi
Current Investigators: Woodruff, Tracey J. , Fisher, Susan J. , McMaster, Michael , Stotland, Naomi , Sutton, Patrice , Gerona, Roy
Institution: University of California - San Francisco
EPA Project Officer: Hahn, Intaek
Project Period: May 5, 2010 through May 4, 2014
Project Period Covered by this Report: May 5, 2011 through May 4,2012
Project Amount: $762,983
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:
R834678C001: Assessing Maternal and Fetal Exposures to Chemicals During the Second Trimester
The objectives of this research remain the same—to generate unprecedented data on chemical exposures during the second trimester of pregnancy, when certain aspects of fetal development are particularly vulnerable to disruption. The specific aims of the project are to (1) compare maternal and fetal exposures to BPA during the second trimester, and (2) explore the feasibility and accuracy of using maternal biological biomonitoring results to predict fetal exposures.
R834678C002: Assessing the Effects of BPA Exposure on Early Human Development
The objective of this research project is to gain an understanding of the effects of BPA exposure on early human development using human embryonic stem cells (hESCs) as a model system. The specific aims are to (1) analyze the impact of BPA exposure on hESC gene expression, and (2) validate the candidate targets and pathways we identify by differentiating BPA-treated hESCs and studying their composition with regard to the progeny of the three definitive germ layers. The specific aims of the project have not been modified.
R834678C003: Predictors of Maternal Exposure to BPA During the Second Trimester of Pregnancy
The objectives of this research remain the same—to understand potential sources of exposure to BPA, and to collect data on the biological half-life of BPA. The specific aims of the project are to (1) investigate the determinants of maternal exposures to BPA during the second trimester of pregnancy, and (2) characterize the change in BPA levels in pregnant women associated with fasting.
R834678C004: Community Outreach and Translation Core (COTC)
- Communicate the meaning of the Pregnancy Exposure to Environmental Contaminants (PEEC) Formative Centers research broadly.
- Create and disseminate a science-based foundation for clinical care and policy change endorsed by key leaders.
- Sustain and expand a collaborative network of partners.
- Advance public policy in the field of reproductive environmental health.
Progress Summary:
R834678C001: Assessing Maternal and Fetal Exposures to Chemicals During the Second Trimester
Over the past year, we have completed study recruitment, worked with our analytical chemist to develop a method for measuring BPA in our serum and urine samples, started preliminary data analysis, and are working on several manuscripts for publication. We also uncovered a source of BPA contamination in the clinic where we are collecting our serum samples, and have identified a BPA-free substitute.
In total, we have recruited 170 women from September 2010 to April 2012. Women eligible for the study attend the clinic over 2 days, and we have collected 125 urines from women for the first day of their visit and 111 urines from women the 2nd day of the visit, which is the day of their medical procedure. From the medical procedure, we have collected 82 umbilical cord bloods, 162 placentas and 153 fetal livers.
Our collaborator in the Department of Laboratory Medicine at San Francisco General Hospital, Dr. Roy Gerona, has developed an improved method for analyzing unconjugated BPA, BPA sulfate and BPA glucuronide. Additionally, during the method development phase, he discovered a source of BPA contamination in our maternal serum samples. He determined the contamination source to be from the BD Vacutainer Push Button Blood Collection Set used in the hospital. We identified a BPA-free needle and started using it on November 29, 2011. We have collected 13 serum samples using the BPA-free needles.
Results:
Among our study, 99 percent of participants have detectable BPA levels in nonfasting urine. The geometric mean concentration is 11.61 ng/mL for BPA glucuronide and 0.98 for BPA sulfate. These levels are slightly higher than levels among pregnant women in previous NHANES cohorts from 2003–2004, possibly due to either our population being primarily low income and more racially diverse, or because we use a method that can directly measure the conjugated BPA. From preliminary data, maternal urine BPA levels (BPA glucuronide and BPA sulfate) have been found to be associated with touching receipts, receipt-related occupations, and consuming more purchased foods outside of the home. Our initial analysis also indicates that receipt-related BPA exposure can come from both occupational and nonoccupational receipt exposure. Further analysis is warranted, but if confirmed, it would provide scientific evidence for an important, yet often neglected, route of exposure. Several other demographic characteristics, such as ethnicity and education, are also found to be associated with maternal BPA levels in our study. As our study population mainly consists of low-income, underserved pregnant women, high BPA levels detected suggest disparities of BPA exposure and likely further differences by other demographic characteristics, such as ethnicity.
For the first time, we provide preliminary measurements of unconjugated and conjugated BPA in umbilical cord serum. We detected unconjugated BPA in 47 percent of umbilical cord serum and 24.4 percent had more unconjugated BPA than conjugated. We find the geometric mean for BPA glucuronide is 0.11 ng/mL and 0.18 ng/mL for BPA sulfate. No other study has provided data on fetal cord blood levels during 2nd trimester, and we have shown the importance of the sulfation metabolic pathway as an alternative to glucornidation.
R834678C002: Assessing the Effects of BPA Exposure on Early Human Development
During the first year of the project, we performed experiments designed to determine that the doses of BPA were effective in modulating gene expression in human embryonic stem cells in the range of exposures documented to occur in pregnant women, and to examine BPA effects on nuclear receptor genes expression. For these experiments, we analysed the UCSF4 cell line, which is included on the Federal Registry (NIH designation 0044). We are adding BPA to hESC cultures in amounts that are comparable to the concentrations that have been measured in pregnancy (~10 ng/mL). We are also including lower (1 ng/mL) and higher (50–100 ng/mL) concentrations. As controls, we add 0.5 nM estradiol (E2) and 0.001 percent EtOH, the vehicle in which BPA was dissolved prior to addition to the medium.
Initial experiments failed to detect any effects of acute BPA exposure for 3 days on hESC self-renewal as monitored by the expression of a set of stage-specific antigens, which we routinely use to monitor pluripotency (e.g., SSEA4, Oct3/4, Nanog, GATA4) and differentiation to the major germ layers—ectoderm, mesoderm and endoderm. Accordingly, we adopted a chronic exposure model in which the cells were cultured for three passages in control and experimental conditions, which is designed to mimic the in vivo situation in which individuals are continuously exposed to environmental chemicals. We monitored the results in terms of mRNA levels for classical steroid receptors (ERα, ERβ, AR), orphan estrogen receptors (ERRα, β, γ), PPARγ, alphafetoprotein (an endoderm marker), Brachyury (a mesodermal marker) and PAX6, which controls the proliferation of neuroepithelial precursors, and stemness markers, including SSEA4 andOct3/4.
qRT-PCR showed that BPA upregulated transcription of genes encoding steroid receptors and steroid responsive genes. These data are in agreement with observed effects of BPA on modulation of nuclear receptors and their target genes in other systems. Markers of hESC differentiation were also modulated by BPA treatment. For example, the brachyury gene encoding a factor important for establishment of the mesodermal lineage in early human embryos was upregulated by BPA treatment. This gene was also highly estrogen responsive. PAX6, a gene important for ectoderm specification and differentiation of neural and epidermal tissues, was downregulated by all doses of BPA tested. In contrast, estrogen had no effect. These and other data suggest that one of the effects of BPA is to disrupt the balance between self-renewal and differentiation is specific populations of cells in the early embryo. Specifically, the results suggest that BPA promotes hESC differentiation toward mesodermal lineages and away from neuroectoderm. In some cases, the effects seem to be mediated through classical steroid response pathways; for other genes, this is not the case.
Recently, we have been focusing on analysis of the effects of BPA on expression of families of transcriptional regulators known to play key roles in hESC self-renewal and differentiation. Hox family members (e.g., HOX-C6, -C9, -B13, -C12, -C10 and DLX2) involved in early morphogenesis are particularly sensitive to regulation by BPA. These and other data support the hypothesis that BPA induces premature differentiation of particular lineages in early embryos. We are currently examining expression of the downstream targets of these transcriptional regulators at the protein level to correlate our morphological observations with the molecular data. We also recently completed the experiments for global transcriptional profiling of hESCs cultured under conditions that induce their differentiation into embryoid bodies. Analysis of this large data set is ongoing. We are discriminating between genes that are affected by BPA and estrogen and those that are differentially regulated, determining the major pathways affected, developing a "molecular fingerprint" for exposure, and analyzing protein expression of regulated targets in the context of hESC self-renewal and differentiation to the major embryonic lineages.
R834678C003: Predictors of Maternal Exposure to BPA During the Second Trimester of Pregnancy
In total, we have collected questionnaire data from 170 study participants. Over the last year, however, we identified that the length of the questionnaire was interfering with study recruitment. Subsequently, we evaluated the responses to the questionnaire and identified questions that were duplicative, or had less than a 10–20 percent response rate. For example, we are no longer asking study participants if they are vegetarian, since none of our participants adhere to a vegetarian diet. We subsequently revised the questionnaire, going from 375 to 197 questions. We collected data on the newer version of the questionnaire from an additional 68 participants. In addition to the questionnaire data, we have also collected 80 matched pairs of fasting and nonfasting maternal urine.
We have also developed a database using REDCap to enter the questionnaire data, and have completed data entry and data validation in accordance with our Quality Management Plan. We have also begun the data analysis phase of this project, producing descriptive statistics on the questionnaire as well as participant demographics collected from clinical chart abstraction records. From the charts, we are collecting information from the counseling records, the pregnancy evaluation, the peri-operative record, and the clinic ultrasound.
Results:
We have collected questionnaire data on 170 participants. Preliminary analysis of questionnaire data was done on a subset of 74 participants. The questionnaire data reflects the socioeconomic status of our study population: 54 percent have high school level education or lower, 25 percent are current smokers, and 54 percent of the participants receive food stamps. We have also found that this study population consumes high quantities of fast foods and other paper-packaged take-out food, with a mean of 7 times/week. Average canned food and canned drink consumption are 5 times/week and 11 times/week, respectively. In terms of knowledge of BPA, 34 percent of the participants have heard about BPA, and 24 percent percent avoid BPA or purchase BPA products.
The questionnaire data along with the BPA levels in maternal urine are being written up as a peer-reviewed publication entitled Dietary, Non-Dietary, and Sociodemographic Predictors of Serum Bisphenol A Body Burden in Pregnant Women. This will be submitted in the summer of 2012. Also, a paper examining the levels in fasting versus non-fasting maternal urine will be completed in the fall of 2012.
R834678C004: COTC
- Hired Jessica Trowbridge at 100 percent to advance our COTC's communication strategy and other COTC activities (June 2011).
- Strengthened our network of collaborators, including convening two meetings of PRHE's From Advancing Science to Ensuring Prevention (FASTEP) Advisory Council (September 2011 and February 2012).
- Developed and implemented a communication strategy to effectively translate the meaning of the science broadly. To this end, we were covered in 45 media stories, including in the San Francisco Chronicle, USA Today online, LA Times, and interviews by KCBS and KCBS Radio; redesigned our website to include better graphics, compatibility with mobile devices, and integration and usability for our collaborators; published four blogs about our scientists, research and related topics in Mission Local, a local online and print newspaper for the San Francisco Mission District; contributed to NIEHS news article "Advising Parents in the Face of Scientific Uncertainty: An Environmental Health Dilemma"; and regularly posted on Twitter and Facebook.
- Advanced the development of a novel methodology to translate the science linking the environment and health in clinical and policy spheres. We published the methodology, the Navigation Guide, in Health Affairs; convened the inaugural meeting of the Navigation Guide Working Group (March 2012); in collaboration with the U.S. Environmental Protection Agency (EPA), initiated the application of the Navigation Guide methodology on a case study of the relationship between exposure to perfluorinated octanoic acid and birth weight; and made numerous presentations to advance the methodology among decision makers in clinical and policy spheres.
- Produced and distributed a wide range of educational materials to support the integration of the science in clinical spheres, including five patient-centered brochures co-branded with FASTEP and other collaborators: Toxic Matters (low-literacy versions in English and Spanish), Pesticides Matter, Work Matters, and Food Matters; produced and distributed Pesticides Matter: A Primer for Reproductive Health Professionals.
- Worked to integrate the science into the purview and activities of the American Congress of Obstetricians & Gynecologists (ACOG) and other health professional societies and institutions. PRHE served as a member of ACOG District IX's (California) legislative committee and as science advisors on reproductive environmental health issues to ACOG District IX and national ACOG; PRHE helped to bring together ACOG and EPA leadership, who in turn identified potential areas of future collaboration; through a mixed methods approach, we gathered the first empirical evidence of knowledge, attitudes and practices related to environmental health among practicing obstetricians in the United States; PRHE served as a member of the California Preconception Care Council; we compiled and disseminated a searchable database of policy statements from health professional organizations related to environmental health: http://prhe.ucsf.edu/prhe/pdfs/Professional%20Statements%20Database.pdf; attended ACOGs national meeting (May 2012); and as listed below, we disseminated the science to health professionals through Grand Rounds and other presentations and publications.
- Collaborated on national efforts to strengthen and build a bridge between the Pediatric Environmental Health Specialty Units (PEHSUs) and obstetricians and their professional organizations in order to ensure a strong referral network for practicing obstetricians who identify a hazardous environmental or occupational exposure.
Future Activities:
R834678C001: Assessing Maternal and Fetal Exposures to Chemicals During the Second Trimester
This project is progressing as expected. We have received data from our urine and cord serum samples, and method development for placenta, fetal liver and amniotic fluid will take place in the coming months. We plan on publishing the results of the method development, as well as data derived from the maternal urine and umbilical cord blood, in the coming months.
R834678C002: Assessing the Effects of BPA Exposure on Early Human Development
The project is progressing as expected. With regard to Aim 1, performance of transcriptional microarrays to determine the effects of BPA exposure on hESC gene expression, we are currently analyzing data from experiments that analyze the effects on the trancriptome of embryoid bodies treated with BPA or estradiol. With regard to Aim 2 experiments, functional analyses of the products of the genes modulated in response to BPA exposure are ongoing.
R834678C003: Predictors of Maternal Exposure to BPA During the Second Trimester of Pregnancy
The questionnaire data, along with the BPA levels in maternal urine, are being written up as a peer-reviewed publication entitled "Dietary, Non-Dietary, and Sociodemographic Predictors of Serum Bisphenol A Body Burden in Pregnant Women." This will be submitted in the summer of 2012. Also, a paper examining the levels in fasting versus nonfasting maternal urine will be completed in the fall of 2012.
Our COTC will continue to collaborate with our wide range of partners through our FASTEP Alliance and Navigation Guide Work Group; communicate the meaning of the science in clinical and policy spheres and to the public though innovative online and social media strategies, presentations and publications; publish the findings of our first case study of the application of the Navigation Guide; publish our findings on knowledge, attitudes and practices related to reproductive environmental health among practicing obstetricians; evaluate the dissemination of our patient-centered brochures; conduct outreach and education activities to harness the voice of health professionals to call for improved environmental health policy; and serve on ACOG District IX's legislative committee and provide input to national ACOG on environmental health science and policy.
Journal Articles: 11 Displayed | Download in RIS Format
| Other center views: | All 73 publications | 15 publications in selected types | All 11 journal articles |
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Gerona RR, Woodruff TJ, Dickenson CA, Pan J, Schwartz JM, Sen S, Friesen MW, Fujimoto VY, Hunt PA. Bisphenol-A (BPA), BPA glucuronide, and BPA sulfate in midgestation umbilical cord serum in a northern and central California population. Environmental Science & Technology 2013;47(21):12477-12485. |
R834678C001 (Final) R834678C003 (Final) |
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Robinson J, Hamilton E, Lam J, Chen H, Woodruff T. Differences in cytochrome p450 enzyme expression and activity in fetal and adult tissues. PLACENTA 2020;100:35-44. |
R834678 (Final) R835433 (Final) |
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Sutton P, Giudice LC, Woodruff TJ. Reproductive environmental health. Current Opinion in Obstetrics and Gynecology 2010;22(6):517-524. |
R834678 (2012) |
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Sutton P, Woodruff TJ, Vogel S, Bero LA. Conrad and Becker's "10 Criteria" fall short of addressing conflicts of interest in chemical safety studies. Environmental Health Perspectives 2011;119(12):A506-507. |
R834678 (2010) R834678 (2011) |
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Sutton P, Wallinga D, Perron J, Gottlieb M, Sayre L, Woodruff T. Reproductive health and the industrialized food system: a point of intervention for health policy. Health Affairs 2011;30(5):888-897. |
R834678 (2010) R834678 (2011) R834678 (2012) |
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Sutton P, Woodruff TJ, Conry J, Giudice LC. Toxic matters and fertility: what can a doctor do? San Francisco Medicine: Journal of the San Francisco Medical Society 2012;85(5)20-22. |
R834678 (2012) |
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Sutton P, Woodruff TJ, Perron J, Stotland N, Conry JA, Miller MD, Giudice LC. Toxic environmental chemicals: the role of reproductive health professionals in preventing harmful exposures. American Journal of Obstetrics and Gynecology 2012;207(3)164-173. |
R834678 (2010) R834678 (2011) R834678 (2012) |
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Sutton P, Woodruff TJ. Risk communication and decision tools for children's health protection. Birth Defects Research Part C: Embryo Today: Reviews 2013;99(1):45-49. |
R834678 (2012) |
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Vandenberg LN, Gerona RR, Kannan K, Taylor JA, van Breemen RB, Dickenson CA, Liao C, Yuan Y, Newbold RR, Padmanabhan V, vom Saal FS, Woodruff TJ. A round robin approach to the analysis of bisphenol A (BPA) in human blood samples. Environmental Health 2014;13(1):25 (20 pp.). |
R834678C001 (Final) R835436 (2014) R835436 (2015) R835436 (2017) |
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Woodruff TJ, Sutton P, Navigation Guide Work Group. An evidence-based medicine methodology to bridge the gap between clinical and environmental health sciences. Health Affairs 2011;30(5):931-937. |
R834678 (2010) R834678 (2011) R834678 (2012) |
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Woodruff TJ, Burke TA, Zeise L. The need for better public health decisions on chemicals released into our environment. Health Affairs 2011;30(5):957-967. |
R834678 (2011) |
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Supplemental Keywords:
Risk assessment, metabolism, bioavailability, ethnic groups, endocrine disruptors, sensitive populations, decision making, measurement methods, RFA, Scientific Discipline, Health, Health Risk Assessment, Biochemistry, Children's Health, Biology, prenatal exposure, biological response, assessment of exposureRelevant Websites:
Advising Parents in the Face of Scientific Uncertainty: An Environmental Health Dilemma
MissionLocal - Clinic Exit Exit
An Ecological Approach to Healthy Aging Across the Lifespan Exit Exit
Navigating the Science: Evaluating Research Quality Exit Exit
Progress 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).
R834678C001 Assessing Maternal and Fetal Exposure to Chemicals
R834678C002 Assessing the Effects of BPA Exposure on Early Human Development
R834678C003 (Pilot Study): Predictors of Maternal Exposure to BPA During Pregnancy
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.