Proteomic analyses of primary human villous trophoblasts exposed to flame retardant BDE-47 using SWATH-MS
Hao Chen1, Katherine E Williams1, Elaine Y Kwan1
1Center for Reproductive Sciences and Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California, San Francisco (UCSF), San Francisco, CA, USA.
Insights
Polybrominated diphenyl ethers (PBDEs) disrupt placental cell development. This study reveals BDE-47 alters protein expression in human cytotrophoblasts, impacting crucial placental functions and potentially leading to adverse birth outcomes.
Area of Science:
- Environmental Toxicology
- Reproductive Biology
- Proteomics
Background:
- Polybrominated diphenyl ethers (PBDEs) are flame retardants with known developmental toxicity.
- PBDEs are detected in placental tissues and linked to adverse birth outcomes.
- Placental cytotrophoblasts (CTBs) are vital for maternal-fetal interface formation and development.
Purpose of the Study:
- To investigate the proteomic changes in human CTBs following exposure to the PBDE BDE-47.
- To identify potential toxicological mechanisms of PBDEs during early placental development.
Main Methods:
- Quantitative proteomic analysis using sequential window acquisition of all theoretical fragment-ion spectra (SWATH-MS).
- Exposure of primary human CTBs to BDE-47 at varying concentrations (1 µM, 5 µM) and time points (15, 24, 39 h).
- Bioinformatic analysis including pathway enrichment and network analysis.
Main Results:
- Over 200 proteins were differentially expressed in CTBs upon BDE-47 exposure, showing time- and concentration-dependent effects.
- Affected proteins were enriched in pathways related to cell aggregation and adhesion.
- The protein CYFIP1 was identified as dysregulated by BDE-47, with potential implications for CTB function.
Conclusions:
- BDE-47 exposure significantly impacts the global proteome of differentiating human CTBs.
- These proteomic alterations suggest mechanisms by which PBDEs may interfere with placental development and function.
- The findings provide a valuable proteomic dataset for understanding environmental chemical impacts on pregnancy outcomes.
Abstract:
Polybrominated diphenyl ethers (PBDEs) are a class of brominated flame retardants and recognized developmental toxicants that are detectable in placental tissues. Higher levels of in utero PBDE exposure have been associated with an increased risk of adverse birth outcomes. During pregnancy, cytotrophoblasts (CTBs) from the placenta play critical roles in the formation of the maternal-fetal interface via uterine invasion and vascular remodeling. The differentiation of these cells towards an invasive phenotype is crucial for proper placental development. We previously have shown that BDE-47 can impact CTB viability and hinder the ability of these cells to migrate and invade. To expand on potential toxicological mechanisms, we utilized quantitative proteomic approaches to identify changes in the global proteome of mid-gestation primary human CTBs after exposure to BDE-47. Using sequential window acquisition of all theoretical fragment-ion spectra (SWATH), we identified 3024 proteins in our CTB model of differentiation/invasion. Over 200 proteins were impacted as a function of BDE-47 exposure (1 μM and 5 μM) across the treatment period (15, 24, and 39 h). The differentially expressed molecules displayed time- and concentration-dependent changes in expression and were enriched in pathways associated with aggregatory and adhesive processes. Network analysis identified CYFIP1, a molecule previously unexplored in a placental context, to be dysregulated at BDE-47 concentrations previously seen to impact CTB migration/invasion. Our SWATH-MS dataset thus demonstrates BDE-47 impacts the global proteome of differentiating CTBs and serves as a valuable resource for further understanding of the relationship between environmental chemical exposures and placental development and function. AVAILABILITY OF DATA AND MATERIAL: Raw chromatograms are deposited on the MassIVE proteomic database (https://massive.ucsd.edu) under accession number MSV000087870. Normalized relative abundances are also available as Table S1.
