Tankyrase inhibition promotes a stable human naïve pluripotent state with improved functionality

• DOI: 10.1242/dev.138982
• Publication Date: 2016-09-23T01:19:17Z
• Authors: Ludovic Zimmerlin; Tea Soon Park; Jeffrey S. Huo; Karan Verma; Sarshan R. Pather; C. Conover Talbot; Jasmin Agarwal; Diana Steppan; Yang W. Zhang; Michael Considine; Hong Guo; Xiufeng Zhong; Christian Gutierrez; Leslie Cope; M. Valeria Canto-...; Alan D. Friedman; Stephen B. Baylin; Elias T. Zambidis
• Keywords: STAT3 Transcription Factor; 0301 basic medicine; Tankyrases; Glycogen Synthase Kinase 3 beta; Induced Pluripotent Stem Cells; Cell Differentiation; Bone Morphogenetic Protein 4; Cellular Reprogramming; Leukemia Inhibitory Factor; 3. Good health; Mice; 03 medical and health sciences; Animals; Humans; Cell Self Renewal; Wnt Signaling Pathway; Cells, Cultured; Embryonic Stem Cells; Germ Layers; Janus Kinases

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.