ESRRB Facilitates the Conversion of Trophoblast-Like Stem Cells From Induced Pluripotent Stem Cells by Directly Regulating CDX2

Abstract

Porcine-induced pluripotent stem cells (piPSCs) could serve as a great model system for human stem cell preclinical research. However, the pluripotency gene network of piPSCs, especially the function for the core transcription factor estrogen-related receptor beta (ESRRB), was poorly understood. Here, we constructed ESRRB-overexpressing piPSCs (ESRRB-piPSCs). Compared with the control piPSCs (CON-piPSCs), the ESRRB-piPSCs showed flat, monolayered colony morphology. Moreover, the ESRRB-piPSCs showed greater chimeric capacity into trophectoderm than CON-piPSCs. We found that ESRRB could directly regulate the expressions of trophoblast stem cell (TSC)-specific markers, including KRT8, KRT18 and CDX2, through binding to their promoter regions. Mutational analysis proved that the N-terminus zinc finger domain is indispensable for ESRRB to regulate the TSC markers. Furthermore, this regulation needs the participation of OCT4. Accordingly, the cooperation between ESRRB and OCT4 facilitates the conversion from pluripotent state to the trophoblast-like state. Our results demonstrated a unique and crucial role of ESRRB in determining piPSCs fate, and shed new light on the molecular mechanism underlying the segregation of embryonic and extra-embryonic lineages.

AXIN2 Reduces the Survival of Porcine Induced Pluripotent Stem Cells (piPSCs)

“…Another huge drawback of the present piPSCs is that it was still hard to contribute to efficient chimeras and was extremely sensitive to perturbation in culture conditions. Previous studies showed that overexpression of BCL2 enhanced piPSCs’ survival under stress and contributed to chimera formation, and this effect was mediated mainly by the increased resistance to apoptosis [ 28 , 29 , 30 ]. In our study, we also confirmed that the caspase family CASPASE3 , CASPASE9 , and apoptotic protein BAX were down-regulated after AXIN2 knockdown.…”

“…To investigate the function of AXIN, the genes, AXIN1 and AXIN2, were PCR-amplified from piPSCs and then were subcloned into EF1-3FLAG-MCS-T2A-puromycin Lentiviral vectors [ 30 ].…”

KDM4C Contributes to Trophoblast-like Stem Cell Conversion from Porcine-Induced Pluripotent Stem Cells (piPSCs) via Regulating CDX2

“…In our recent study, we found that ESRRB could trigger piPSC differentiation into TLSCs through activating CDX2 and KRT8 expression [ 5 ]. However, the mechanisms of histone modifications underlying the conversion process are yet to be established.…”

“…Recently, Io et al established an in vitro model of the trophoblast lineage from human iPSCs [ 4 ]. We also found that Estrogen-Related Receptor Beta (ESRRB) overexpression in pig iPSCs (piPSCs) could cause the trans-differentiation of piPSCs into TLSCs [ 5 ]. This result reflects the powerful function of the transcription factor ESRRB.…”

Enhancement of Chromatin and Epigenetic Reprogramming in Porcine SCNT Embryos—Progresses and Perspectives

“…Overexpression of the iPSC inducing factors OCT-4 , SOX2 , KLF4 and c-MYC (OSKM) in porcine cells prior to nuclear transfer improved blastocyst rates and quality of SCNT embryos ( Song Z. et al, 2014 ; Kim et al, 2019 ). The co-expression of OKSM and the estrogen-related receptor B ( ESRRB ), which is abundantly expressed in pig embryos ( Yu et al, 2021 ), improved iPSCs production by regulating pluripotency factors ( Shi et al, 2020 ). In addition, pig iPSCs overexpressing ESRRB showed higher potential for trophectoderm differentiation when injected into 8-cell stage embryos ( Yu et al, 2021 ).…”