Grantee Research Project Results

Our project tests the hypotheses that 1) polybrominated diphenyl ethers (PBDEs), perfluorinated chemicals (PFCs), and select environmental organic acids (EOAs) accumulate differently in human fetuses than in pregnant women during the 2nd trimester of pregnancy, and 2) these chemical exposures negatively impact placental development and function, an important outcome that can adversely affect in utero growth, prenatal development, birth weight, and thus, childhood and adult health. Compelling scientific data show that human in utero development is highly susceptible to disruption by exposure to chemicals, in particular endocrine disrupting agents, but a major impediment to understanding these risks is the lack of human data on fetal exposures and on the mechanisms by which chemicals can adversely impact in utero development. Through our work as a Formative Center, we provided some of the first human data characterizing midgestation chemical exposures and mechanisms of adverse effects.

Specifically, we detected PBDEs, PFCs and EOAs such as Bisphenol-A (BPA) in 99-100% of the pregnant women and 95- 100% of fetuses (liver and umbilical cord blood) in our study population. In addition, in vitro experiments that employed our novel human trophoblast progenitor cell (TBPC) model of placental development demonstrated that environmentally relevant doses of PBDE-47 (a PBDE congener) compromised TBPC self-renewal in an undifferentiated state while promoting differentiation (see Project 1, Aim 1). Both effects are hallmarks of poor placental development and have been shown to be risk factors for reduced fetal growth, preterm delivery and preeclampsia. Our project will significantly advance this promising line of research to include a larger sample size, a more diverse set of biological specimens and a wider range of measured chemicals. We will also test the impact of these chemicals on in utero development by assessing the relationship between direct measures of chemical exposure and morphological and molecular markers that enable an assessment of placentation. Accordingly, our specific aims are to:

• Compare maternal serum, placenta and fetal liver levels of a broad range of chemicals through traditional and novel biomonitoring approaches. We will collect matched biological specimens from 130 pregnant women obtaining elective 2nd trimester pregnancy terminations (excluding cases in which pregnancies are complicated by fetal anomalies or maternal smoking). We will analyze the samples for 17 PBDEs and 12 PFCs. We will also use a novel biomonitoring approach based on Time-of- Flight Liquid Chromatography-Mass Spectrometry (LC-QTOF/MS) combined with traditional analytic methods to identify and quantify exposure to the 5 most frequently detected EOAs in our study population. We will compare measurements of these 34 chemicals in maternal serum to those in the placenta and fetal liver. Thus, at the conclusion of these experiments we will have a much broader understanding of the most significant environmental chemical exposures during pregnancy and the relationships between levels detected in the maternal blood, placenta and fetus.
• Evaluate the relationship between placental disruption and exposure to PBDEs, PFCs and select EOAs. We will measure known morphological and molecular markers of placenta development in the 2nd trimester placentas that are collected in Aim 1. Then we will evaluate the relationship of these markers to individual and cumulative metrics of PBDE, PFC and EOA exposure measured in the same placental samples. Additionally, the goal of Project 1 is to identify novel PBDE and PFC placental targets (specifically, PBDE-47 and PFOA, respectively). In Aim 3 of that Project, we will also evaluate the relationship between these endpoints, which will be analyzed in the same placental samples, and individual and cumulative metrics of PBDE, PFC and EOA exposure. Thus, at the conclusion of these experiments we will have important new information about the placental effects of endocrine disrupting chemicals.

This project will examine the hypothesis that prenatal exposures to endocrine disrupting chemicals (EDCs) affect fetal growth outcomes such as birth weight and small for gestational age (SGA), and whether these adverse effects are magnified by chronic psychosocial stress. We will conduct a prospective study in an economically and ethnically diverse group of pregnant women in order to evaluate two important types of environmental exposures: 1) EDCs - polybrominated diphenyl ethers (PBDEs) and perfluorinated chemicals (PFCs); and 2) chronic psychosocial stress, using objective and subjective measures of participants’ perceived stress, social standing, physical and social environments, and biomarkers of chronic stress response. We hypothesize that exposure to both EDCs and stress may have synergistic effects on fetal growth. Accordingly, this project has three specific aims:

Research Project I: Modeling the Effects of EDCs on Early Stages of Human Placental Development

During the fifth project period, we continued to make progress and summarize/finalize results related to Aims 1-3. In this project, we are using a primary human cytotrophoblast (CTB) model system in which we have over 20 years of experience to study placentation in normal pregnancy and in pregnancy complications. We also selected this primary placental cell model to leverage recently generated transcriptomic and epigenomic data due to our participation in the NIH-funded Roadmap Epigenome Mapping Center. From these studies, we gained a unique understanding of the transcriptomic and epigenetic landscape of CTBs and other extraembryonic tissues in vivo under normal physiological conditions. This detailed information provides us a baseline for understanding molecular perturbations that result from exposure to environmental chemicals, the goal of this project.

With regard to Aims 1 and 2, we conducted transcriptomic and methylation analyses of the effects of BDE-47 exposure in our CTB model system. Based on our dose/time response investigations completed in Year 1 and 2 which established relevant timing and dosage parameters in our model, we conducted transcriptomic and epigenomic analyses in Year 3, which included: 1) a sub-cytotoxic concentration of BDE-47 (1µM) which was biologically-effective in our model system and relevant to human exposures; and 2) two exposure windows of interest in cultured CTBs, which paralleled significant dynamic changes in cell migratory activity and expression of genes linked to morphogenesis, cell motility, and vascular remodeling pathways. These fundamental cellular and molecular processes are hypothesized to underlie the critical functions of CTBs in vivo, such as uterine invasion, vascular remodeling, and establishing robust physical and physiological connections between the placenta and uterus.

In CTBs, we examined responses to BDE-47 exposures after 24h due to reports suggesting PBDEs do alter gene/protein expression in trophoblast cell lines at equivalent exposure durations (2) and our initial dose-response investigations in Years 1 and 2. We collected RNA/DNA samples simultaneously from 3 independent cultures of 2nd trimester human CTBs which were exposed to either BDE-47 (1 µM) or vehicle control (1% DMSO; 12 total samples for each analysis). To evaluate gene expression, we used Affymetrix GeneChip Human Gene 2.0 ST Arrays, which contains probes for over 30,000 coding transcripts. For epigenomic analyses, we measured global methylation using Illumina Infinium HumanMethylation450 BeadChip arrays. This approach enabled us to investigate methylation levels for ~99% of genes and ~96% of CpG islands in the human genome. In Years 4-5, we finalized the analysis of these datasets and experimental studies aimed at confirming changes on the RNA/protein level of specific targets. Following data normalization, we applied a fixed effects linear model (ANOVA) to determine differentially expressed (DE) genes due to BDE-47 exposure, culture time, and gestational age. Our previous experiments in (unexposed) cultured CTBs and in vivo suggest that culture-time and gestational age influences the expression of hundreds of genes important for placental development and function, therefore, we used a statistical approach that controls for these variables. In total, we identified 276 genes to be significantly differentially expressed (DE) due to BDE-47 exposure (ANOVA: p ≤ 0.025, absolute FC > 1.25). In general, expression changes due to exposure of BDE-47 were similar between the two 24h- exposure intervals despite CTBs being exposed at two different timepoints (3 vs. 15h post-plating), with 100% of DE genes displaying common trends in response to BDE-47 (y = 0.98x, R2 = 0.81). In total, 159 (58%) genes were upregulated and 117(42%) genes were downregulated due to BDE-47 exposure. The majority of enriched biological processes were driven by upregulated transcripts. They included morphogenesis, vasculature development, cell differentiation, cell migration, signal transduction, inflammatory response, protein metabolism, and regulation of biosynthetic process- related pathways. GO terms related to lipid and steroid metabolism were overrepresented by downregulated targets. We identified several novel genes dysregulated associated with PBDE exposure. Using qRT-PCR, we confirmed targets across a BDE-47 concentration range of 0.1-10µM. We observed significant upregulation of IL6, MMP1, GREM1, FABP4, and PLAC4; and downregulation of GPR34, SCD, HMGCS1 and FABP7 with BDE-47 (p < 0.05). These validated targets represent key members of diverse functional pathways involved in placental development, including: trophoblast differentiation, inflammation, fatty acid metabolism, steroid metabolism and/or morphogenesis. Minimal changes were observed at concentrations ≤ 0.1µM. In summary, our transcriptomic results suggest BDE-47, in a concentration dependent manner, to perturb known and undefined pathways linked with environmental toxicity and placental development.

In parallel with global mRNA analyses, we profiled the CpG methylome of BDE-47 (1µM) and vehicle exposed CTBs following a 24h duration (also initiated at 3h or 15h post-plating). All data were processed using a state-of-the-art pipeline includes methylumi background correction and normalization operations. We identified 758 CpGs as differentially methylated (DM) with BDE-47 exposure (p ≤ 0.005; change ≥ 2.5%). In general, BDE-47 increased global CpG methylation (0.8 ± 0.2%), heightened when the analysis was constrained to BDE-47 DM CpGs (3.4 ± 0.3%). Ninety-three percent of BDE-47 DM CpGs had increased methylation (708/758 total). On average, methylation changes due to BDE-47 were similar whether exposures were initiated at 3h or 15h (y=0.90x, R2=0.69). Within the BDE-47 DM CpG subset, we identified underrepresentation of CpGs proximal to promoter regions (e.g., TSS1500, TS200, 1st Exon) and enriched for intergenic regions. GO analyses of genes near BDE-47 DM CpGs indicated enrichment of pathways involved in hormone response, cellular projection, signaling, response to oxygen-containing compounds, morphogenesis, and vesicle transport/localization. We identified an enrichment of DM CpGs on chromosomes 10 and 11 and a relative absence on chromosome 18. These analyses pointed us towards finding an interesting DM CpG cluster on Ch. 11p15.5 known to play a critical role in gestational development and poised to be sensitive to environmental exposures. Interestingly, in general, BDE-47- induced changes in DNA methylation did not correlate with mRNA expression levels. In total, only four genes (3 unique mRNAs) whose promoters were DM had mRNA levels that were also responsive to BDE-47.  These initial -omic studies provide us information regarding our experimental design and provide clues to prepare for more resource-intensive investigations that will be initiated in the near future as we expand this model for placental toxicological investigations.

As related to Aim 3, we developed tests to evaluate 1) migratory activity or 2) the ability of cells to invade using enhanced workflows that include fixation, nuclear staining/immunofluorescence, multiple image capture (Leica), and semi-automated data content analysis (Volocity). We added automated components to classical methodologies to reduce human error and increase efficiency in assessing functional endpoints across multiple conditions. We determined key timepoints in culture by conducting time-course evaluations of migratory activity and cell invasion. Using the standard 2-D culture system described above, in unexposed cultures, we quantified migratory activity/aggregation of CTBs within the first 40h of culture by measuring the average distance between > 20,000 cells per well as a parameter of activity. To assess cell invasion, we cultured cells in a suspended transwell (Corning) insert covered with Matrigel and auto-counted (Volocity) the total number of cellular projections penetrating the substrate that appear on the underside of the permeable filter. Similar to migration assays, we observed significant invasion within the first 40h of culture, suggesting that the invasive/migratory programming that these cells undergo in vivo is conserved in culture. We are currently finalizing the data in which we assessed BDE-47 or -99 exposed vs. control CTBs. In our initial experiments, cell invasion was inhibited at concentrations of 5μM BDE-47 or -99. These assays will enable us to link mechanistic relationships between environmental exposures and morphological phenotypic behaviors relevant to the in vivo condition. Furthermore, these endpoints will be combined with the other morphological (time-lapse imaging) and molecular (immunolocalization, immunoblot, qRT-PCR) assessments that we have used to characterize environmental (i.e., BDE-47 and PFOA) effects in CTBs. Aim 3 experiments are designed to analyze the functions of molecules that are targets of the ECs of interest and which we suspect play important roles in human placental development. The deep understanding we are developing of the normal transcriptomic and epigenomic landscape of CTBs will help put the data we generate in the exposure experiments in the proper context and inform future targeted analyses.

To understand the relevancy of concentrations tested and determine the degree of bioaccumulation and metabolism of BDE-47 in our CTB model system, we quantified BDE-47 and hydroxylated metabolites of the parent species, i.e., 5-OH-BDE-47, 6-OH-BDE-47, in CTBs (cell and media fractions) exposed to BDE- 47 via liquid chromatography–mass spectrometry (LC-MS). The results indicated that BDE-47 significantly bioaccumulated in the cellular fraction, in a concentration-dependent manner. After 24h, BDE-47 concentrations were 0.7µM or 1.2µM in the supernatant of cells initially exposed to 1µM or 5µM, respectively. Additionally, we computed the total µg recovered of BDE-47 in the media and cellular fractions. BDE-47 media (µg) to cell (µg) ratios differed between the two dose groups (1 vs 5µM). In cultures exposed to 1µM BDE-47, we observed a cell:media ratio of 0.6:1µg. In cultures exposed to 5µM, BDE-47 partitioned between the cell and media fractions at 3.1:1µg. These results suggest bioaccumulation of BDE-47 in the cellular fraction, and the rate of cellular uptake to be dependent on the magnitude of the initial testing concentration. Total recovery (media + cellular fractions) were within 10% of expected estimates. Limited metabolism of BDE-47 was observed in CTBs exposed to 1 or 5µM, with only ~0.0003% of the total BDE-47 recovered (media and cellular fractions) present in the hydroxylated form. Furthermore, only the 6-OH-BDE-47 metabolite (not the 5-OH), was identified in the cellular fraction in one of the three CTB samples evaluated. In addition to BDE-47, four of the eighteen PBDE congeners evaluated, i.e., -17, 28, -85, -99, were detected at low levels in the majority of all samples, representing < 0.4% of the ∑PBDEs. No form of BDE-47 was present in any samples exposed to the vehicle control. Overall, these results indicate that exposures in this study consisted predominately of the BDE-47 parent compound and only limited hydroxylated metabolism occurred in CTBs within the first 24h of exposure.

We re-submitted a manuscript entitled “Genomic Profiling of BDE-47 Effects on Human Placental Cytotrophoblasts.” This manuscript describes a global transcriptional analysis of time-dependent changes in gene expression occurring in primary CTBs as they differentiate in vitro. This study serves as a baseline for our environmental investigations. We will present our transcriptomic analyses of BDE-47 and complementary results in an abstract: "Toxicogenomic Profiling of BDE-47 Effects in Human Primary Villous Cytotrophoblasts”, at the Teratology Society Annual Meeting in Denver, CO in June, 2017. Dr. Robinson has also presented results from these analyses in several forums at UCSF, including for the Department of Obstetrics, Gynecology & Reproductive Sciences, “Grand Rounds”, and Center for Reproductive Sciences, “Annual Retreat”.

Building on our accomplishments from FY1–FY4, we also achieved a number of accomplishments for Project 2 in FY5:

• Methods for PBDE analysis in serum and liver were developed by our collaborators at California Department of Toxic Substances (DTSC), Drs. Myrto Petreas and June-Soo Park. Methods development for PBDE analysis in placenta was also finalized. In December of 2016, we completed PBDE chemical analysis on all 130 matched sets of maternal serum, placenta, and fetal liver.

• Over the past year, our post-doc, Julia Varshavsky, completed a correlation analysis evaluating PBDE level across biological matrices, including maternal serum, the placenta, and fetal liver tissues. Statistical methods included linear regression, the non-parametric Kendall’s tau correlation, and methods that account for left-censored chemical exposure data. She presented this work at the joint annual meeting of the International Society of Exposure Science and the International Society of Environmental Epidemiology which took place August 26-30, 2018. Julia will be drafting a manuscript of the results from this analysis in the coming weeks that we expect to submit in October 2018. In addition, we have combined the maternal serum PBDE data from the N = 130 paired samples with previous maternal serum PBDE data from women who were also recruited at the Women’s Options Clinic in San Francisco General Hospital between 2007 and 2012 in order to evaluate geographical, temporal, and racial/ethnic differences across a larger study population.

• Placenta samples were fixed in formalin for future immunohistochemistry analyses. Based on findings in Project I, which evaluated the effects of BDE-47 on functional and transcriptomic levels in 2nd trimester human primary cytotrophoblasts, potential biomarkers of PBDE were identified. We selected four of these targets to further interrogate as potential biomarkers of PBDE exposure in vivo, which included, MMP11-3 and PLAC44, molecules with suspected roles in placental development; and HMGCS15 and SCD6, members of the cholesterol/fatty acid biosynthesis pathway. We also added proposed biomarkers of pre-eclampsia (GH2, PLAC1) and negative controls/molecules not altered by PBDEs in vitro (NOTUM, EFEMP1) to our analyses. Using antigen-specific antibodies, we independently evaluated the localized expression of each of these targets within a subset of placentas (n = 12), pre-identified to be in the upper (> 10%) or bottom (> 90%) percentiles of ∑PBDE burden within our sample set. Two sections from each placenta were probed and scored blindly by two investigators to determine the relative expression of each marker using a semi-automated method that we developed to quantify relative immunofluorescence intensity within regions of the maternal-fetal interface. We analyzed six regions in total: 1) CTBs invading into the decidua (interstitial invasion); 2) decidual cells; 3) CTBs of the proximal and distal column, 4) villous trophoblasts (STBs and CTBs), 5) mesenchymal cells within the floating villous, and 6) CTBs invading the maternal artery (endovascular invasion). All sections were co-stained with cytokeratin (CK), a marker of CTB populations, and 4', 6-diamidino-2-phenylindole (Dapi), a DNA nuclear binding molecule. Specific patterns were observed. For example, SCD was expressed highest in the floating villous CTBs, whereas PLAC4 was predominately expressed in the villous core mesenchyme, interstitial CTBs, and decidua. Relative intensity values for each of the eight biomarkers were summarized. We observed good agreement between investigators (r=0.8) and across technical replicates (r=0.7). Staining patterns varied across individual placentas.

In fund Year 5 (FY) we finished recruiting participants at Zuckerberg San Francisco General Hospital (ZSFG), and the Betty Irene Moore Women’s Health Hospital at Mission Bay (MB). We exceeded our recruitment goal by achieving a final recruitment number of 510 participants in August of 2017.

PBDE methods development in serum has been completed by our collaborators at California Department of Toxic Substances Drs. Myrto Petreas and June-Soo Park; all maternal serum samples have been transported to the DTSC for PBDEs analysis The first batch of PDBEs analysis (n = 237) was completed in September of 2017. The remaining 276 samples will be completed in the fall of 2018.

The Fisher lab completed CRH analysis on 434 maternal plasma samples in 2018. Also, the Blackburn Lab completed telomere analysis on approximately 300 cord whole blood samples and in September 2018, we will receive the telomere results for our final ~30 cord blood samples.

We hired two post-doctoral fellows, Drs. Monika Izano, an epidemiologist from UC Berkeley’s School of Public Health, and Elizabeth Hom, from the University of Washington’s School of Public Health, both of whom have been collaborating with our team to conduct data analysis and manuscript preparation during this project year. This year we had two abstracts accepted for presentation at the 2018 ISEE/ISES conference in Ottawa, Canada. One abstract, to be presented as a poster, examined effects of maternal psycho-social stress on newborn telomere length (TL) and gestational age. This preliminary analysis applied three statistical approaches, traditional regression analysis, combined with data driven, agnostic models including variable importance analysis, and TMLE, to examine these relationships. VIM analysis found that stressors predicted more verifiability in telomere length and gestational age, compared to biological factors and measures of maternal socioeconomic status, TMLE results suggested relationships between maternal perceived stress exposure depressions associated with shorter TL, while job strain and discrimination were associated with longer TL. We will assess the robustness of these results once receive the rest of the telomere data among our participants. The second abstract, accepted as an oral presentation, examined associations between PFAS and biomarkers of chronic stress response applying similar traditional and agnostic statistical techniques. We found significant associations between PFNA and total PFAS levels and maternal CRH levels, with chemicals explaining more of the variance in CRH than either SES or maternal biological factors in the VIM analysis. However, we did not see any association between maternal PFAS levels and telomere length in any of our statistical models. With support from an Administrative Supplement, we collected 50 placentas from women in our CIOB2 cohort. In Project 1 of our UCSF PEEC Children's Center, we have developed an in vitro model of human placentation to study the effects of PBDEs and PFCs on trophoblasts—the basic building blocks of this organ—at transcriptomic, epigenomic and functional levels. In Project 2, we are integrating exposure science with placental biology by using targeted and non-targeted biomonitoring methods to measure human maternal and fetal exposures to PBDEs, PFCs and

Aim 2: Harness the Evolving Science to Healthcare: Inquiry: In August 2017 we were invited to create and implement an environmental health course as part of the “Inquiry” deep-dive component of the Bridges curriculum – a mandatory education for all 2nd year medical students. Our team was subsequently asked to create another Inquiry course for August 2018, which will include Dr. Woodruff providing a foundational lecture on the importance of Environmental Health and a case study on the impacts of environmental exposures on infertility, adapted from The Story of Health created by the Pediatric Environmental Health Specialty Unit (PEHSU).

Elective: In fall 2017 we developed and implemented the “Women, the Environment, and Physician Activism: Encouraging Activism through Education” elective for students in the schools of medicine, nursing, pharmacy, and dentistry, and will again implement the elective in fall 2018. We have offered 6 elective courses since 2013, including 4 in the School of Medicine and 2 in collaboration with the School of Pharmacy. Since its inception, between 300-400 students have taken this course and 14 medical students have lead its planning.

LifeStages syllabus: In summer 2017 we revised the LifeStages syllabus into a “Reader” format for the August 2017 lecture, which compiled the material in a more accessible manner for students. In fall 2017, we partnered with Mark Miller and Stephanie Holm from the Western States Pediatric Environmental Health Specialty Unit (PEHSU) to develop a clinically-focused model for integrating environmental health content into all topic areas of the LifeStages medical education curriculum reader. LifeStages is the course material of the new Bridges Medical Education Program that contains all of the lectures and related readings for the Reproductive health, Pediatric, and Geriatric subjects the 2nd year medical students are taught. Working with a 4th year medical student, we used the model to develop the environmental health content for the reader to generate content for 10 syllabus topics/lectures initially focused in reproductive health. Our collaboration with Washington State University resulted in an invited presentation about our medical education work at the 10th World Congress Alternatives and Animal Use in the Life Sciences conference in August 2017: http://wc10seattle.org/2017/General- Information.aspx

Building on many of the relationships we developed through the UC Carbon Neutrality Initiative, which produced a core group of academics and students across UCSF who are working to embed environmental health into their area of the curriculum, we are growing the Environmental Health Initiative and expanding collaborations across the Schools of Medicine, Nursing, Pharmacy, and Dentistry on the topic of environmental health in healthcare education and practice. Two of our UC Carbon Neutrality Initiative (CNI) fellows, Carolyn Rennels and Gabriela Weigel, were responsible for organizing and implementing the “Women, the Environment, and Physician Activism: Encouraging Activism through Education” elective referenced above.

Aim 3: Advance Prevention‐Based Public Policy: Our goal to improve the use of science in decision making to achieve safer building materials and consumer product has been significantly advanced by our collaboration with Touro University. Our plans to leverage current data on formaldehyde and asthma to a broader set of outcomes, especially cancer, will be influential in decision-making on common building materials used in affordable housing: composite wood products. The U.S. Environmental Protection Agency (EPA) considered cancer outcomes in its Formaldehyde Emissions Standards for Composite Wood Products, which has been subject to litigation and unnecessary controversy. Furthermore, how to address financial conflicts of interest in scientific research remains an unresolved issue. In addition to building the evidence base on formaldehyde, the ultimate product of this work will inform general systematic review methodology and incorporation of financial conflict of interest information in evaluating the quality of research studies. Our collaboration with a new partner, Touro University, has been a tremendous success, generating training opportunities in systematic review methods with faculty and students. Developing this new pipeline of scientists is instrumental to ensuring the implementation and evolution of systematic review methods. As we look forward to the completion of this project, we are in active discussion with Touro to further our work with new systematic review projects. This project is contributing to our long-term outcomes and overall objectives through the training of the future generation of scientists in systematic review methods, which will increase the uptake of systematic and transparent methods for reaching evidence-based decisions in environmental health. Our work on "Double Jeopardy," i.e., cumulative co-exposure to social stressors such as poverty and toxic environmental chemicals, has been presented by Dr. Rachel Morello-Frosch (“Cumulative prenatal exposures to exogenous chemicals and psychosocial stress”) speaking at the EPA/NIEHS Children’s Centers Annual Meeting in Oct 2017 and Dr. Tracey Woodruff presenting in Dec 2017 at an event convened in collaboration with UCSF Benioff Children’s Hospital: “Conversations on Children’s Health: Toxic Chemicals and Toxic Stress.” As the science on cumulative risk continues to develop, we also need evaluation methods that incorporate the most current science on cumulative exposures, stressors and impacts. To establish the state of the field on such methods, we published a paper reviewing modeling methods utilized to quantify the cumulative effects of multiple stressors in previous studies. We blogged about our results and promoted on social media, providing policy context and recommendations for a policy-focused audience. (References: Huang H, Wang A, Morello-Frosch R, Lam J, Sirota M, Padula A, et al. Cumulative Risk and Impact Modeling on Environmental Chemical and Social Stressors. Curr Environ Heal Reports. 2018 Feb 13;5(1):88–99. Blog: 1+1>2: Evaluating how risks of pollutants and stressors stack up https://prheucsf.blog/2018/03/05/112-evaluating-how-risks-of-pollutants-and-stressors-stack-up/).

We have continued to implement our systematic review training curriculum with academic, government, NGO and industry scientists and researchers. We tested the training materials: 1) online to a group of 30 scientists who are currently doing systematic reviews for a World Health Organization (WHO)-International Labor Organization (ILO) joint project; 2) a group of 100 scientists at the Society of Toxicology (SOT) annual meeting in San Antonio, Texas; 3) a group of 75 scientists at the University of Michigan (online and in person); 4) a group of 40 US EPA employees representing virtually every Office within the Agency who are responsible for conducting and implementing systematic review methodology; 5) one scientist at Health Canada who the lead for a project to implement the Navigation Guide systematic review methodology for chemicals assessments at the Agency; and 6) one scientist at the Western Suburbs of Paris University Hospital working on a novel application of Navigation Guide --evaluating occupational health case reports. We have found that the demand for training in systematic review methods is quite high, as is the value of improving capabilities to assess the scientific evidence and come to evidence-based conclusions, which will lead to substantial benefits to protecting the public’s health. In addition to training the pipeline of scientists directly with our curriculum, we are providing expertise, insight, and feedback on the conduct of several systematic review projects. This includes: 1) a case study conducted at Texas A&M University on air pollution and adverse birth outcomes; 2) 12 case studies implemented in a joint project between WHO/ILO on occupational health-related topics; 3) a case study at the Western Suburbs of Paris University Hospital using Navigation Guide at the intersection of medical science and environmental health to evaluate occupational health case reports; and 4) a case study conducted at Touro University on the impact of financial conflicts of interest on study quality and author conclusions regarding toxicity. These collaborations and projects contribute to our long-term outcomes and overall objectives through the increasing the uptake of systematic and transparent methods for reaching evidence-based decisions among environmental health scientists, in both the academic and government arena.

Core B: Administrative Core

External Committee and EAC Meeting

The PEEC External Advisory Committee (EAC) held three project-based meetings in 2018. .

Project 1 & 2 held a joint meeting with three EAC members on May 29, 2018. The goal of this meeting was to discuss progress and next steps for Projects 1 & 2 in the final year of funding, specifically pertaining to evaluating biological pathways and endpoints of maternal/fetal chemical exposures.

The COTC met with EAC members on February 27, 2018 with two EAC members. The goal of the COTC meeting was two-fold: 1) to share accomplishments (and challenges) of the Clinical Outreach and Translation Core from these past 5 years and 2) to benefit from expertise and feedback in solidifying our “niche” and goals for the next 5-10 years.

During each EAC meetings, the projects and the COTC delivered a comprehensive presentation to bring attention to project accomplishments, current obstacles, and future directions, which was followed by a lengthy discussion with EAC members to advise project leads on changes that could be made in the future, and suggestions for possible follow-up grants based on the results of this work.

Our next step is to implement the content we have developed for the 10 syllabus topics, which we plan to begin in fall 2018. To be successful we will need the support and endorsement of both the faculty overseeing their respective area of LifeStages (Preconception/reproductive health, Pediatrics, and Geriatrics) as well as the lecturers of each course. To this end, we have the support and endorsement of Dr. Naomi Stotland, our colleague who oversees the Preconception/Reproductive health section of LifeStages medical school curriculum, as she has also been our champion and advocate for greater integration of environmental health in the medical education curriculum. We have also recently garnered the support of Dr. Patrick Newman, the faculty member overseeing the Pediatrics element of LifeStages.