Induction of a Human Pluripotent State with Distinct Regulatory Circuitry that Resembles Preimplantation Epiblast

Abstract

Human embryonic stem cells (hESCs) are derived from the inner cell mass of the blastocyst. Despite sharing the common property of pluripotency, hESCs are notably distinct from epiblast cells of the preimplantation blastocyst. Here we use a combination of three small-molecule inhibitors to sustain hESCs in a LIF signaling-dependent hESC state (3iL hESCs) with elevated expression of NANOG and epiblast-enriched genes such as KLF4, DPPA3, and TBX3. Genome-wide transcriptome analysis confirms that the expression signature of 3iL hESCs shares similarities with native preimplantation epiblast cells. We also show that 3iL hESCs have a distinct epigenetic landscape, characterized by derepression of preimplantation epiblast genes. Using genome-wide binding profiles of NANOG and OCT4, we identify enhancers that contribute to rewiring of the regulatory circuitry. In summary, our study identifies a distinct hESC state with defined regulatory circuitry that will facilitate future analysis of human preimplantation embryogenesis and pluripotency.

Naïve Induced Pluripotent Stem Cells Generated From β-Thalassemia Fibroblasts Allow Efficient Gene Correction With CRISPR/Cas9

“…4A). Gene ontology [18] enrichment analysis showed that the upregulated genes in naïve iPSCs were mainly involved in translation, embryonic development, and oxidative phosphorylation (Fig. 4D).…”

“…Previous studies have also suggested that human primed iPSCs and mouse EpiSCs can be converted to the naïve ground state via chemical manipulation or the overexpression of specific transcription factors [12][13][14][15][16][17]. Recently, several groups have further modified culture conditions to maintain human ESCs or iPSCs in the naïve ground state [12,13,15,18]. In particular, the 5i/L/FA system, developed by Jaenisch's group, contains a combination of inhibitors and growth factors sufficient for the induction and maintenance of the naïve ground state [15].…”

The molecular underpinnings of totipotency

“…These conditions also make ES cell derivation possible from normally recalcitrant mouse strains [33] and rat embryos [34], and most recently a variant of 2i has been used to derive naive human ES cells [15][16][17]. In 2i conditions, ES cells are also heterogeneous and include a significant fraction of cells that can be prospectively isolated based on the expression of the extraembryonic transcription factor Hhex [35] (figure 1b).…”

T-box 3 overexpression is associated with poor prognosis of non-small cell lung cancer

“…Accumulating evidence has demonstrated that Tbx3 is involved in the developmental process by binding to the T-box motif with a 20-24 nucleotide palindromic sequence, the T-site, or half of the sequence in the promoters of target genes (5). A mutation of Tbx3 was demonstrated to be closely associated with the pluripotency of embryonic stem cells and the invasiveness of cancer (6). In resectable pancreatic carcinoma (7), melanoma (8), bladder cancer (9) and colorectal cancer (10), Tbx3 was revealed to be overexpressed and associated with poor prognosis of cancer.…”