title: 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.
authors: Gene Regulation Laboratory, Genome Institute of Singapore, Singapore 138672, Singapore. Electronic address: chanysw@gis.a-star.edu.sg.
authors: Yun-Shen
authors: YS
authors: Chan
authors: Jonathan
authors: J
authors: Göke
authors: Jia-Hui
authors: JH
authors: Ng
authors: Xinyi
authors: X
authors: Lu
authors: Kevin Andrew Uy
authors: KA
authors: Gonzales
authors: Cheng-Peow
authors: CP
authors: Tan
authors: Wei-Quan
authors: WQ
authors: Tng
authors: Zhong-Zhi
authors: ZZ
authors: Hong
authors: Yee-Siang
authors: YS
authors: Lim
authors: Huck-Hui
authors: HH
authors: Ng
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000187:drug effects / Q000378:metabolism*; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D044127:Epigenesis, Genetic / Q000187:drug effects; D050993:GATA6 Transcription Factor / Q000378:metabolism; D053263:Gene Regulatory Networks / Q000187:drug effects; D005855:Germ Layers / Q000187:drug effects / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D000090062:Kruppel-Like Factor 4; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology; D051379:Mice; D008954:Models, Biological; D000071317:Nanog Homeobox Protein; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D054852:Small Molecule Libraries / Q000494:pharmacology; D059467:Transcriptome / Q000187:drug effects / Q000235:genetics
chemical_list: D050993:GATA6 Transcription Factor; C494759:GATA6 protein, human; D018398:Homeodomain Proteins; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D053605:Leukemia Inhibitory Factor; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D054852:Small Molecule Libraries