Cited by 165Pluripotent Stem Cells ResearchDevelopmental Biology and Gene RegulationEpigenetics and DNA Methylation

Principles of signaling pathway modulation for enhancing human naive pluripotency induction

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Bayerl, Jonathan, Ayyash, Muneef, Shani, Tom, Manor, Yair Shlomo, Gafni, Ohad, Massarwa, Rada, Kalma, Yael, Aguilera-Castrejon, Alejandro +21 more·2021·Cell stem cell

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Abstract

Abstract Different conditions have been devised to isolate MEK/ERK signalling independent human naïve pluripotent stem cells (PSCs) that are distinct from conventional primed PSCs and better correspond to pre-implantation developmental stages. While the naïve conditions described thus far endow human PSCs with different extents of naivety features, isolating human pluripotent cells that retain characteristics of ground state pluripotency while maintaining differentiation potential and genetic integrity, remains a major challenge. Here we engineer reporter systems that allow functional screening for conditions that can endow both the molecular and functional features expected from human naive pluripotency. We establish that simultaneous inhibition of SRC-NFκB, WNT/ßCATENIN and PKC signalling pathways is essential for enabling expansion of teratoma competent fully naïve human PSCs in defined or xeno-free conditions. Divergent signalling and transcriptional requirements for maintaining naïve pluripotency were found between mouse and human. Finally, we establish alternative naïve conditions in which MEK/ERK inhibition is substituted with inhibition for NOTCH/RBPj signalling, which allow obtaining alternative human naïve PSCs with diminished risk for loss of imprinting and deleterious global DNA hypomethylation. Our findings set a framework for the signalling foundations of human naïve pluripotency and may advance its utilization in future translational applications. Highlights of key findings Combined inhibition of SRC, WNT and PKC signaling consolidates human naïve pluripotency Stable expansion of DNA/RNA methylation-independent and TGF/ACTIVIN-independent human naïve PSCs Opposing roles for ACTIVIN and WNT/ßCATENIN signaling on mouse vs. human naive pluripotency 2i and MEK/ERKi independent alternative human naïve PSC conditions via inhibiting NOTCH/RBPj signaling

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Crossref · 10.1016/j.stem.2021.04.001

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  • TCA cycle activity varies with PSC identity in human ESCs, which display glutamine independence and increased mitochondrial respiration upon conversion to a more naïve state.

    “TCA cycle activity also varies with PSC identity in human ESCs, which display glutamine independence and increased mitochondrial respiration upon conversion to a more naïve state (215, 216).”

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Authors
Bayerl, J., Ayyash, M., Shani, T., Manor, Y. S., Gafni, O., Massarwa, R., Kalma, Y., Aguilera-Castrejon, A., Zerbib, M., Amir, H., Sheban, D., Geula, S., Mor, N., Weinberger, L., Naveh Tassa, S., Krupalnik, V., Oldak, B., Livnat, N., Tarazi, S., Tawil, S., Wildschutz, E., Ashouokhi, S., Lasman, L., Rotter, V., Hanna, S., Ben-Yosef, D., Novershtern, N., Viukov, S., & Hanna, J. H.
Affiliations
  • Weizmann Institute of Science (IL)
  • Tel Aviv University (IL)
  • Rambam Health Care Campus (IL)
Corresponding author
  • Bayerl, Jonathan
  • Novershtern, Noa
  • Hanna, Jacob H.
Venue
Cell stem cell
Publisher
Elsevier BV
Published
2021-09
Volume
28
Issue
9
Pages
1549-1565.e12
ISSN
  • 1875-9777 (ISSN-L)
  • 1934-5909
OpenAlex
W3159163097
Cited by
165
References
83
Citations by year
2026: 24 · 2025: 36 · 2024: 29 · 2023: 39 · 2022: 26 · 2021: 9 · 2020: 1 · 2015: 1
FWCI
8.08
Type
journal article
Language
en
Open access
Locations
  • Cell stem cell (publishedVersion, cc-by-nc-nd)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
Funders
  • Israel Cancer Research Fund
  • Flight Attendant Medical Research Institute
  • Minerva Foundation
  • Israel Science Foundation
  • New York Stem Cell Foundation
  • BSF
  • European Research Council (ERC-CoG-2016 726497)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Epigenetics and DNA Methylation
Keywords
naive pluripotency, human pluripotent stem cells, MEK/ERK signaling, Wnt/β-catenin signaling, protein kinase C signaling, Src signaling, trophoblast stem cells, extraembryonic endoderm, p53, DNA hypomethylation
MeSH terms
Animals, Cell Differentiation, Embryo, Mammalian, Humans, Trophoblasts, Signal Transduction, Pluripotent Stem Cells, Mice

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