Cross-sectional associations between prenatal maternal per- and poly-fluoroalkyl substances and bioactive lipids in three Environmental influences on Child Health Outcomes (ECHO) cohorts

Background: Per- and poly-fluoroalkyl substances (PFAS) exposure can occur through ingestion of contaminated food and water, and inhalation of indoor air contaminated with these chemicals from consumer and industrial products. Prenatal PFAS exposures may confer risk for pregnancy-related outcomes such as hypertensive and metabolic disorders, preterm birth, and impaired fetal development through intermediate metabolic and inflammation pathways. Objective: Estimate associations between maternal pregnancy PFAS exposure (individually and as a mixture) and bioactive lipids. Methods: Our study included pregnant women in the Environmental influences on Child Health Outcomes Program: Chemicals in our Bodies cohort (CiOB, n=73), Illinois Kids Developmental Study (IKIDS, n=287), and the ECHO-PROTECT cohort (n=54). We measured twelve PFAS in serum and 50 plasma bioactive lipids (parent fatty acids and eicosanoids derived from cytochrome p450, lipoxygenase, and cyclooxygenase) during pregnancy (median 17 gestational weeks). Pairwise associations across cohorts were estimated using linear mixed models and meta-analysis. Associations between the PFAS mixture and individual bioactive lipids were estimated using quantile g-computation. Results: PFDeA, PFOA, and PFUdA were associated (p<0.05) with changes in bioactive lipid levels in all three enzymatic pathways (cyclooxygenase [n=6 signatures]; cytochrome p450 [n=5 signatures]; lipoxygenase [n=7 signatures]) in at least one combined cohort analysis. The strongest signature indicated that a doubling in PFOA corresponded with a 24.3% increase (95% CI [7.3%, 43.9%]) in PGD2 (cyclooxygenase pathway) in the combined cohort. In the mixtures analysis, we observed nine positive signals across all pathways associated with the PFAS mixture. The strongest signature indicated that a quartile increase in the PFAS mixture was associated with a 34% increase in PGD2 (95% CI [8%, 66%]), with PFOS contributing most to the increase. Conclusions: Bioactive lipids were revealed as biomarkers of PFAS exposure and could provide mechanistic insights into PFAS' influence on pregnancy outcomes, informing more precise risk estimation and prevention strategies. ### Competing Interest Statement The authors declare they have no competing financial or non-financial interests as defined by Environment International, or other interests that might be perceived to influence the results and/or discussion reported in this paper. ### Funding Statement Research reported in this publication was supported by the Environmental Influences on Child Health Outcomes (ECHO) Program, Office of the Director, National Institutes of Health (NIH), under award numbers U2COD023375 (Coordinating Center), U24OD023382 (Data Analysis Center), U24OD023319 with co-funding from the Office of Behavioral and Social Science Research (Pro-Core), U2COD023375 (Opportunities and Infrastructure Fund), UH3OD023251 (Alshawabkeh), UH3OD023272 (Schantz and Woodruff). This work was additionally supported by the Children's Environmental Health and Disease Prevention Research Center (grant numbers ES022848 and RD83543401), the Molecular Phenotyping and Metabolomics Core, Michigan Nutrition and Obesity Center (5P30DK089503), US Environmental Protection Agency (grant number RD83543301), National Institute of Environmental Health Sciences (NIEHS; grant numbers P42ES017198, P50ES026049, P01ES022841, P30ES019776, P30ES030284, and P30ES007048). M.T.A and S.M.E. were also supported in part by the JPB Foundation Environmental Health Fellows Program. The content of this study is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: Properly constituted Institutional Review Boards, either the ECHO single IRB or the ECHO cohort's local IRB, are accountable for compliance with regulatory requirements for the ECHO-wide Cohort Data Collection Protocol at participating cohort sites. Governing IRBs review ECHO protocols and all informed consent, assent forms, HIPAA authorization forms, recruitment materials, and other relevant information prior to the initiation of any ECHO wide Cohort Data Collection Protocol related procedures or activities. ECHO Cohort Investigators (or their designated study personnel) obtain written informed consent or parent's or guardian's permission along with child assent as appropriate, for ECHO wide Cohort Data Collection Protocol participation and for participation in their specific cohorts. The work of the ECHO Data Analysis Center is approved through the Johns Hopkins Bloomberg School of Public Health Institutional Review Board. I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes R-script used for the present study are found in supplemental materials. Select de-identified data from the ECHO Program are available through NICHD's Data and Specimen Hub (DASH). Information on study data not available on DASH, such as some Indigenous datasets, can be found on the ECHO study DASH webpage.