Comparative Study of PPARγ Targets in Human Extravillous and Villous Cytotrophoblasts

Abstract

Trophoblasts, as the cells that make up the main part of the placenta, undergo cell differentiation processes such as invasion, migration, and fusion. Abnormalities in these processes can lead to a series of gestational diseases whose underlying mechanisms are still unclear. One protein that has proven to be essential in placentation is the peroxisome proliferator-activated receptor γ (PPARγ), which is expressed in the nuclei of extravillous cytotrophoblasts (EVCTs) in the first trimester and villous cytotrophoblasts (VCTs) throughout pregnancy. Here, we aimed to explore the genome-wide effects of PPARγ on EVCTs and VCTs via treatment with the PPARγ-agonist rosiglitazone. EVCTs and VCTs were purified from human chorionic villi, cultured in vitro, and treated with rosiglitazone. The transcriptomes of both types of cells were then quantified using microarray profiling. Differentially expressed genes (DEGs) were filtered and submitted for gene ontology (GO) annotation and pathway analysis with ClueGO. The online tool STRING was used to predict PPARγ and DEG protein interactions, while iRegulon was used to predict the binding sites for PPARγ and DEG promoters. GO and pathway terms were compared between EVCTs and VCTs with clusterProfiler. Visualizations were prepared in Cytoscape. From our microarray data, 139 DEGs were detected in rosiglitazone-treated EVCTs (RT-EVCTs) and 197 DEGs in rosiglitazone-treated VCTs (RT-VCTs). Downstream annotation analysis revealed the similarities and differences between RT-EVCTs and RT-VCTs with respect to the biological processes, molecular functions, cellular components, and KEGG pathways affected by the treatment, as well as predicted binding sites for both protein-protein interactions and transcription factor-target gene interactions. These results provide a broad perspective of PPARγ-activated processes in trophoblasts; further analysis of the transcriptomic signatures of RT-EVCTs and RT-VCTs should open new avenues for future research and contribute to the discovery of possible drug-targeted genes or pathways in the human placenta.

Prostaglandin E2 Receptor 4 (EP4) Affects Trophoblast Functions via Activating the cAMP-PKA-pCREB Signaling Pathway at the Maternal-Fetal Interface in Unexplained Recurrent Miscarriage

“…In our project, we explored the molecular mechanisms behind the regulation of the PGE2 receptor EP4 in trophoblasts in first trimester placentas. Liu et al suggested that PGE2 modulates biochemical and morphological placental trophoblast differentiation during implantation and placentation [17]. Previous research demonstrated that placental hypoxia activated cAMP production, proliferation, and invasion by mediating cAMP-PKA-pCREB signaling in trophoblast cells, predominantly through EP1 and EP4 [18,19].…”

“…Supernatants were collected from 24-well plates of HTR-8/SVneo, JEG-3, and BeWo cells that had been incubated in defined medium for 24 and 36 h, respectively, and refluxed at 10,000× g for 10 min for filtration of cell sections. β-hCG and progesterone were gauged using the recommended guidelines with an ADVIA Centaur XP auto analyzer (Siemens Medical Solution Diagnostics, Erlangen, Germany), described in previous studies [17].…”

Understanding bovine embryo elongation: a transcriptomic study of trophoblastic vesicles

“…Different effects were analyzed to characterize their variability including tissue types and gestational days. Data processing further used the methods detailed in Liu et al (2020) . In brief, the threshold to identify the up- and downregulated genes per statistical comparison was set to a fold change of 1.5, and the Gene Ontology (GO) terms (or KEGG pathways) associated with the differential expressions (all DEGs, as well as up- or downregulated DEGs) were recovered using ClueGO.…”