An integrated atlas of human placental development delineates essential regulators of trophoblast stem cells

Abstract

The trophoblast lineage safeguards fetal development by mediating embryo implantation, immune tolerance, nutritional supply and gas exchange. Human trophoblast stem cells (hTSCs) provide a platform to study lineage specification of placental tissues; however, the regulatory network controlling self-renewal remains elusive. Here, we present a single-cell atlas of human trophoblast development from zygote to mid-gestation together with single-cell profiling of hTSCs. We determine the transcriptional networks of trophoblast lineages in vivo and leverage probabilistic modelling to identify a role for MAPK signalling in trophoblast differentiation. Placenta- and blastoid-derived hTSCs consistently map between late trophectoderm and early cytotrophoblast, in contrast to blastoid-trophoblast, which correspond to trophectoderm. We functionally assess the requirement of the predicted cytotrophoblast network in an siRNA-screen and reveal 15 essential regulators for hTSC self-renewal, including MAZ, NFE2L3, TFAP2C, NR2F2 and CTNNB1. Our human trophoblast atlas provides a powerful analytical resource to delineate trophoblast cell fate acquisition, to elucidate transcription factors required for hTSC self-renewal and to gauge the developmental stage of in vitro cultured cells.

Resolving the gene expression maps of human first-trimester chorionic villi with spatial transcriptome

“…These TFs were also highly expressed in EVT in our results ( Figure 4B ). We also confirmed the dominant expressions and spatial distribution of AHR, CEBPA, TBX3 and NFE2L3 in the nuclei of SCT ( Figures 4B,C ), consistent with previous studies ( Simmons et al, 2008 ; Iqbal et al, 2021 ; Chen et al, 2022b ; Dong et al, 2022 ). GATA6, ZEB1, RUNX1, and ERG are fibroblast or endothelial cells-related TFs ( Molkentin, 2000 ; Ponder et al, 2016 ; Yuan et al, 2020 ; Zhang et al, 2021b ; Caporarello et al, 2022 ).…”

Single-cell assessment of trophoblast stem cell-based organoids as human placenta-modeling platforms

“…Gene regulatory network analyses identified TFs driving key trophoblast progenitor cell regulons. We confirm the importance of known trophoblast progenitor transcripts both in vivo and in our TOrgs, including GATA2/3 41, 42 , TFAP2A/C 41, 42 , TEAD4 31–33 , and the newly defined ARID3A and TEAD1 43 . In addition, our findings highlight novel TFs (i.e., EGR1 , TFAP2B , FOXP1 , FOXO4 , IRF6 ) regulating CTB biology in vivo and in TOrgs.…”

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

“…In addition, an improved understanding of the interplay between placental genes and endometrial environmental cues that are essential for hTSC specification and differentiation may aid in understanding the genetic and environmental basis of placental pathologies leading to fetal growth restriction, preterm birth, and recurrent pregnancy loss [120][121][122], while genes that suppress the proliferation and invasion of hTSCs may provide candidates for treatment of choriocarcinoma, a highly malignant tumor of trophoblastic origin [123]. From this standpoint, recent candidate-based and genome-wide approaches to identify essential and growth-restricting genes in hTSCs present an important foundation for future studies [124][125][126][127][128]. We anticipate that the ensuing decade will witness unprecedented advances in our understanding of placental development and women's health through the concerted efforts of reproductive scientists, stem cell biologists, clinicians, and biomedical engineers.…”