Department of Developmental Biology and Cancer Research, The Institute for Medical Research Israel-Canada, The Hebrew University-Hadassah Medical School, 91120, Jerusalem, Israel.
Department of Stem Cell Bioengineering, Mossakowski Medical Research Institute, Polish Academy of Sciences, Warsaw, 02-106, Poland.
Nat Commun. 2023 Jun 8;14(1):3359. doi: 10.1038/s41467-023-39104-1.
Human trophoblast stem cells (hTSCs) can be derived from embryonic stem cells (hESCs) or be induced from somatic cells by OCT4, SOX2, KLF4 and MYC (OSKM). Here we explore whether the hTSC state can be induced independently of pluripotency, and what are the mechanisms underlying its acquisition. We identify GATA3, OCT4, KLF4 and MYC (GOKM) as a combination of factors that can generate functional hiTSCs from fibroblasts. Transcriptomic analysis of stable GOKM- and OSKM-hiTSCs reveals 94 hTSC-specific genes that are aberrant specifically in OSKM-derived hiTSCs. Through time-course-RNA-seq analysis, H3K4me2 deposition and chromatin accessibility, we demonstrate that GOKM exert greater chromatin opening activity than OSKM. While GOKM primarily target hTSC-specific loci, OSKM mainly induce the hTSC state via targeting hESC and hTSC shared loci. Finally, we show that GOKM efficiently generate hiTSCs from fibroblasts that harbor knockout for pluripotency genes, further emphasizing that pluripotency is dispensable for hTSC state acquisition.
人滋养层干细胞（hTSC）可从胚胎干细胞（hESC）中衍生而来，也可通过 OCT4、SOX2、KLF4 和 MYC（OSKM）从体细胞中诱导产生。在这里，我们探讨了 hTSC 状态是否可以在不依赖于多能性的情况下诱导产生，以及获得该状态的机制是什么。我们发现 GATA3、OCT4、KLF4 和 MYC（GOKM）是一种组合因子，可以从成纤维细胞中产生功能性 hiTSC。稳定的 GOKM 和 OSKM-hiTSC 的转录组分析显示，有 94 个 hTSC 特异性基因在 OSKM 衍生的 hiTSC 中特异性异常。通过时间过程 RNA-seq 分析、H3K4me2 沉积和染色质可及性，我们证明 GOKM 比 OSKM 具有更大的染色质开放性活性。虽然 GOKM 主要靶向 hTSC 特异性基因座，但 OSKM 主要通过靶向 hESC 和 hTSC 共享基因座来诱导 hTSC 状态。最后，我们表明 GOKM 可以有效地从携带多能性基因敲除的成纤维细胞中产生 hiTSC，这进一步强调了多能性对于获得 hTSC 状态是可有可无的。
