The microRNA cluster C19MC confers differentiation potential into trophoblast lineages upon human pluripotent stem cells

Abstract

The first cell fate commitment during mammalian development is the specification of the inner cell mass and trophectoderm. This irreversible cell fate commitment should be epigenetically regulated, but the precise mechanism is largely unknown in humans. Here, we show that naïve human embryonic stem (hES) cells can transdifferentiate into trophoblast stem (hTS) cells, but primed hES cells cannot. Our transcriptome and methylome analyses reveal that a primate-specific miRNA cluster on chromosome 19 (C19MC) is active in naïve hES cells but epigenetically silenced in primed ones. Moreover, genome and epigenome editing using CRISPR/Cas systems demonstrate that C19MC is essential for hTS cell maintenance and C19MC-reactivated primed hES cells can give rise to hTS cells. Thus, we reveal that C19MC activation confers differentiation potential into trophoblast lineages on hES cells. Our findings are fundamental to understanding the epigenetic regulation of human early development and pluripotency.

Partial Disturbance of Microprocessor Function in Human Stem Cells Carrying a Heterozygous Mutation in the DGCR8 Gene

“…However, it is currently unknown why this heterozygous mutation did not show a 100% penetrance in our experiments. One explanation could lie in the naïve versus primed stage of the used human embryonic stem cell line: in a recent study, it was revealed that the C19MC is relatively highly expressed in naïve hESCs, whereas its transcriptional activity is declined in primed stem cells [ 53 ]. In our culturing conditions, the HuES9 ESC line rather contains primed cell populations, and the lower expression of the miRNA cluster in those cell types might hinder the detection of the position effect in pri-miRNA processing.…”

Accessing the human trophoblast stem cell state from pluripotent and somatic cells

“…Naïve hPSCs give rise to a transient pre-implantation TE state when cultured in BMP4, Activin/Nodal inhibitor A83-01, MEK inhibitor PD03, and a Janus Kinase inhibitor. TE cells can then be transitioned to stable post-implantation hTSCs in SAVECY or ACE, a reduced version of SAVECY [29,30] primed, hPSCs display complete activation of placentally imprinted C19MC miRNAs [105], which is one of the original trophoblast criteria proposed by Lee et al [38]. Ectopic activation of C19MC miRNAs in primed hPSCs enhanced their potential to differentiate into proliferative hTSCs with full differentiation potential towards specialized trophoblast lineages [105].…”

“…However, there may yet be subtle but significant differences between hTSC-like cells derived from naïve and primed states. For example, a recent study from the Arima laboratory reported that hTSCs derived from naïve, but not primed, hPSCs display complete activation of placentally imprinted C19MC miRNAs [ 105 ], which is one of the original trophoblast criteria proposed by Lee et al [ 38 ]. Ectopic activation of C19MC miRNAs in primed hPSCs enhanced their potential to differentiate into proliferative hTSCs with full differentiation potential towards specialized trophoblast lineages [ 105 ].…”

“…TE cells can then be transitioned to stable post-implantation hTSCs in SAVECY or ACE, a reduced version of SAVECY [29,30] primed, hPSCs display complete activation of placentally imprinted C19MC miRNAs [105], which is one of the original trophoblast criteria proposed by Lee et al [38]. Ectopic activation of C19MC miRNAs in primed hPSCs enhanced their potential to differentiate into proliferative hTSCs with full differentiation potential towards specialized trophoblast lineages [105]. Consistent with these findings, incomplete activation of C19MC miRNAs was also observed in two other studies that derived hTSCs from primed hPSCs [101,103].…”

Role of microRNAs in trophoblast invasion and spiral artery remodeling: Implications for preeclampsia

“…In human embryonic stem (hES) cells, C19MC expression is important for the differentiation of hES into human trophoblast stem cells (hTS). Using CRISPR/Cas gene editing tools, the authors further demonstrate that C19MC is critical in hTS maintenance ( Kobayashi et al, 2022 ). The exact mechanism behind C19MC stem cell maintenance is uncertain but it may involve the C19MC-induced upregulation of pluripotent factors octamer-binding transcription factor 4 ( OCT4 ) and fibroblast growth factor 4 ( FGF4 ) ( Mong et al, 2020 ).…”