title: Tankyrase inhibition promotes a stable human naïve pluripotent state with improved functionality.
abstract: The derivation and maintenance of human pluripotent stem cells (hPSCs) in stable naïve pluripotent states has a wide impact in human developmental biology. However, hPSCs are unstable in classical naïve mouse embryonic stem cell (ESC) WNT and MEK/ERK signal inhibition (2i) culture. We show that a broad repertoire of conventional hESC and transgene-independent human induced pluripotent stem cell (hiPSC) lines could be reverted to stable human preimplantation inner cell mass (ICM)-like naïve states with only WNT, MEK/ERK, and tankyrase inhibition (LIF-3i). LIF-3i-reverted hPSCs retained normal karyotypes and genomic imprints, and attained defining mouse ESC-like functional features, including high clonal self-renewal, independence from MEK/ERK signaling, dependence on JAK/STAT3 and BMP4 signaling, and naïve-specific transcriptional and epigenetic configurations. Tankyrase inhibition promoted a stable acquisition of a human preimplantation ICM-like ground state via modulation of WNT signaling, and was most efficacious in efficiently reprogrammed conventional hiPSCs. Importantly, naïve reversion of a broad repertoire of conventional hiPSCs reduced lineage-primed gene expression and significantly improved their multilineage differentiation capacities. Stable naïve hPSCs with reduced genetic variability and improved functional pluripotency will have great utility in regenerative medicine and human disease modeling.
authors.affiliation: Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.|Division of Pediatric Oncology, Baltimore, MD 21205, USA.
authors.forename: Ludovic
authors.identifier: 0000-0002-6031-0348
authors.initials: L
authors.lastname: Zimmerlin
authors.affiliation: Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.|Division of Pediatric Oncology, Baltimore, MD 21205, USA.
authors.forename: Tea Soon
authors.identifier: 0000-0002-2519-1291
authors.initials: TS
authors.lastname: Park
authors.affiliation: Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.|Division of Pediatric Oncology, Baltimore, MD 21205, USA.
authors.forename: Jeffrey S
authors.identifier: 0000-0002-0057-9118
authors.initials: JS
authors.lastname: Huo
authors.affiliation: Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.|Division of Pediatric Oncology, Baltimore, MD 21205, USA.
authors.forename: Karan
authors.initials: K
authors.lastname: Verma
authors.affiliation: Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.|Division of Pediatric Oncology, Baltimore, MD 21205, USA.
authors.forename: Sarshan R
authors.identifier: 0000-0001-5959-2096
authors.initials: SR
authors.lastname: Pather
authors.affiliation: Institute for Basic Biomedical Sciences at Johns Hopkins, Baltimore, MD 21205, USA.
authors.forename: C Conover
authors.initials: CC
authors.lastname: Talbot
authors.affiliation: Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.|Division of Pediatric Oncology, Baltimore, MD 21205, USA.
authors.forename: Jasmin
authors.initials: J
authors.lastname: Agarwal
authors.affiliation: Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.|Division of Pediatric Oncology, Baltimore, MD 21205, USA.
authors.forename: Diana
authors.initials: D
authors.lastname: Steppan
authors.affiliation: Division of Cancer Biology, Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Baltimore, MD 21205, USA.
authors.forename: Yang W
authors.initials: YW
authors.lastname: Zhang
authors.affiliation: Division of Cancer Biology, Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Baltimore, MD 21205, USA.
authors.forename: Michael
authors.identifier: 0000-0002-4468-6820
authors.initials: M
authors.lastname: Considine
authors.affiliation: Division of Pediatric Oncology, Baltimore, MD 21205, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Guo
authors.affiliation: Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
authors.forename: Xiufeng
authors.initials: X
authors.lastname: Zhong
authors.affiliation: Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
authors.forename: Christian
authors.initials: C
authors.lastname: Gutierrez
authors.affiliation: Division of Cancer Biology, Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Baltimore, MD 21205, USA.
authors.forename: Leslie
authors.initials: L
authors.lastname: Cope
authors.affiliation: Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
authors.forename: M Valeria
authors.initials: MV
authors.lastname: Canto-Soler
authors.affiliation: Division of Pediatric Oncology, Baltimore, MD 21205, USA.
authors.forename: Alan D
authors.initials: AD
authors.lastname: Friedman
authors.affiliation: Division of Cancer Biology, Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Baltimore, MD 21205, USA.
authors.forename: Stephen B
authors.identifier: 0000-0003-3697-3798
authors.initials: SB
authors.lastname: Baylin
authors.affiliation: Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA ezambid1@jhmi.edu.|Division of Pediatric Oncology, Baltimore, MD 21205, USA.
authors.forename: Elias T
authors.identifier: 0000-0001-5452-254X
authors.initials: ET
authors.lastname: Zambidis
mesh_terms: D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D000066673:Cell Self Renewal / Q000502:physiology*; D002478:Cells, Cultured; D065150:Cellular Reprogramming / Q000502:physiology; D053595:Embryonic Stem Cells / Q000166:cytology*; D005855:Germ Layers / Q000196:embryology; D000071679:Glycogen Synthase Kinase 3 beta / Q000037:antagonists & inhibitors; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D053612:Janus Kinases / Q000378:metabolism; D053605:Leukemia Inhibitory Factor / Q000378:metabolism; D051379:Mice; D050796:STAT3 Transcription Factor / Q000378:metabolism; D038501:Tankyrases / Q000037:antagonists & inhibitors*; D060449:Wnt Signaling Pathway / Q000502:physiology*
chemical_list: C525809:BMP4 protein, human; D055415:Bone Morphogenetic Protein 4; C507837:LIF protein, human; D053605:Leukemia Inhibitory Factor; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D038501:Tankyrases; D053612:Janus Kinases; D000071679:Glycogen Synthase Kinase 3 beta
keywords: Differentiation; Ground state; Human embryonic stem cell; Induced pluripotent stem cell; Naïve pluripotency