Induction of a human pluripotent state with distinct regulatory circuitry that resembles preimplantation epiblast

Yun-Shen Chan; Jonathan Göke; Jia-Hui Ng; Xinyi Lu; Kevin Andrew Uy Gonzales; Cheng-Peow Tan; Wei-Quan Tng; Zhong-Zhi Hong; Yee-Siang Lim; Huck-Hui Ng

doi:10.1016/j.stem.2013.11.015

Induction of a human pluripotent state with distinct regulatory circuitry that resembles preimplantation epiblast

Cell Stem Cell. 2013 Dec 5;13(6):663-75. doi: 10.1016/j.stem.2013.11.015.

Authors

Yun-Shen Chan¹, Jonathan Göke, Jia-Hui Ng, Xinyi Lu, Kevin Andrew Uy Gonzales, Cheng-Peow Tan, Wei-Quan Tng, Zhong-Zhi Hong, Yee-Siang Lim, Huck-Hui Ng

Affiliation

¹ Gene Regulation Laboratory, Genome Institute of Singapore, Singapore 138672, Singapore. Electronic address: chanysw@gis.a-star.edu.sg.

PMID: 24315441
DOI: 10.1016/j.stem.2013.11.015

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Blastocyst / drug effects
Blastocyst / metabolism*
Cell Line
Cell Proliferation / drug effects
Embryonic Stem Cells / cytology
Embryonic Stem Cells / drug effects
Embryonic Stem Cells / metabolism
Epigenesis, Genetic / drug effects
GATA6 Transcription Factor / metabolism
Gene Regulatory Networks* / drug effects
Germ Layers / drug effects
Germ Layers / metabolism*
Homeodomain Proteins / metabolism
Humans
Kruppel-Like Factor 4
Leukemia Inhibitory Factor / pharmacology
Mice
Models, Biological
Nanog Homeobox Protein
Pluripotent Stem Cells / cytology*
Pluripotent Stem Cells / drug effects
Pluripotent Stem Cells / metabolism*
Signal Transduction / drug effects
Signal Transduction / genetics
Small Molecule Libraries / pharmacology
Transcriptome / drug effects
Transcriptome / genetics

Substances

GATA6 Transcription Factor
GATA6 protein, human
Homeodomain Proteins
KLF4 protein, human
Klf4 protein, mouse
Kruppel-Like Factor 4
Leukemia Inhibitory Factor
NANOG protein, human
Nanog Homeobox Protein
Small Molecule Libraries