Principles of signaling pathway modulation for enhancing human naive pluripotency induction

Jonathan Bayerl; Muneef Ayyash; Tom Shani; Yair Shlomo Manor; Ohad Gafni; Rada Massarwa; Yael Kalma; Alejandro Aguilera-Castrejon; Mirie Zerbib; Hadar Amir; Daoud Sheban; Shay Geula; Nofar Mor; Leehee Weinberger; Segev Naveh Tassa; Vladislav Krupalnik; Bernardo Oldak; Nir Livnat; Shadi Tarazi; Shadi Tawil; Emilie Wildschutz; Shahd Ashouokhi; Lior Lasman; Varda Rotter; Suhair Hanna; Dalit Ben-Yosef; Noa Novershtern; Sergey Viukov; Jacob H. Hanna

doi:10.1016/j.stem.2021.04.001

Principles of signaling pathway modulation for enhancing human naive pluripotency induction

Jonathan Bayerl^*, Muneef Ayyash, Tom Shani, Yair Shlomo Manor, Ohad Gafni, Rada Massarwa, Yael Kalma, Alejandro Aguilera-Castrejon, Mirie Zerbib, Hadar Amir, Daoud Sheban, Shay Geula, Nofar Mor, Leehee Weinberger, Segev Naveh Tassa, Vladislav Krupalnik, Bernardo Oldak, Nir Livnat, Shadi Tarazi, Shadi TawilEmilie Wildschutz, Shahd Ashouokhi, Lior Lasman, Varda Rotter, Suhair Hanna, Dalit Ben-Yosef^*, Noa Novershtern^*, Sergey Viukov, Jacob H. Hanna^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

75 Scopus citations

Abstract

Isolating human MEK/ERK signaling-independent pluripotent stem cells (PSCs) with naive pluripotency characteristics while maintaining differentiation competence and (epi)genetic integrity remains challenging. Here, we engineer reporter systems that allow the screening for defined conditions that induce molecular and functional features of human naive pluripotency. Synergistic inhibition of WNT/β-CATENIN, protein kinase C (PKC), and SRC signaling consolidates the induction of teratoma-competent naive human PSCs, with the capacity to differentiate into trophoblast stem cells (TSCs) and extraembryonic naive endodermal (nEND) cells in vitro. Divergent signaling and transcriptional requirements for boosting naive pluripotency were found between mouse and human. P53 depletion in naive hPSCs increased their contribution to mouse-human cross-species chimeric embryos upon priming and differentiation. Finally, MEK/ERK inhibition can be substituted with the inhibition of NOTCH/RBPj, which induces alternative naive-like hPSCs with a diminished risk for deleterious global DNA hypomethylation. Our findings set a framework for defining the signaling foundations of human naive pluripotency.

Original language	English
Pages (from-to)	1549-1565.e12
Journal	Cell Stem Cell
Volume	28
Issue number	9
DOIs	https://doi.org/10.1016/j.stem.2021.04.001
State	Published - 2 Sep 2021

Funding

Funders	Funder number
Kekst Family Institute for Medical Genetics , Helen and Martin Kimmel Institute for Stem Cell Research
Kekst Family Institute for Medical Genetics, Helen and Martin Kimmel Institute for Stem Cell Research
Israel Cancer Research Fund
Flight Attendant Medical Research Institute
European Research Council	ERC-CoG-2016 726497
European Research Council
Minerva Foundation
United States-Israel Binational Science Foundation
Israel Science Foundation

Keywords

cross-species chimerisim
embryonic stem cells
extra-embryonic stem cells
iPSC
naive pluripotency
reprogramming

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.stem.2021.04.001

Cite this

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., ... Hanna, J. H. (2021). Principles of signaling pathway modulation for enhancing human naive pluripotency induction. Cell Stem Cell, 28(9), 1549-1565.e12. https://doi.org/10.1016/j.stem.2021.04.001

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title = "Principles of signaling pathway modulation for enhancing human naive pluripotency induction",

abstract = "Isolating human MEK/ERK signaling-independent pluripotent stem cells (PSCs) with naive pluripotency characteristics while maintaining differentiation competence and (epi)genetic integrity remains challenging. Here, we engineer reporter systems that allow the screening for defined conditions that induce molecular and functional features of human naive pluripotency. Synergistic inhibition of WNT/β-CATENIN, protein kinase C (PKC), and SRC signaling consolidates the induction of teratoma-competent naive human PSCs, with the capacity to differentiate into trophoblast stem cells (TSCs) and extraembryonic naive endodermal (nEND) cells in vitro. Divergent signaling and transcriptional requirements for boosting naive pluripotency were found between mouse and human. P53 depletion in naive hPSCs increased their contribution to mouse-human cross-species chimeric embryos upon priming and differentiation. Finally, MEK/ERK inhibition can be substituted with the inhibition of NOTCH/RBPj, which induces alternative naive-like hPSCs with a diminished risk for deleterious global DNA hypomethylation. Our findings set a framework for defining the signaling foundations of human naive pluripotency.",

keywords = "cross-species chimerisim, embryonic stem cells, extra-embryonic stem cells, iPSC, naive pluripotency, reprogramming",

author = "Jonathan Bayerl and Muneef Ayyash and Tom Shani and Manor, {Yair Shlomo} and Ohad Gafni and Rada Massarwa and Yael Kalma and Alejandro Aguilera-Castrejon and Mirie Zerbib and Hadar Amir and Daoud Sheban and Shay Geula and Nofar Mor and Leehee Weinberger and {Naveh Tassa}, Segev and Vladislav Krupalnik and Bernardo Oldak and Nir Livnat and Shadi Tarazi and Shadi Tawil and Emilie Wildschutz and Shahd Ashouokhi and Lior Lasman and Varda Rotter and Suhair Hanna and Dalit Ben-Yosef and Noa Novershtern and Sergey Viukov and Hanna, {Jacob H.}",

note = "Publisher Copyright: {\textcopyright} 2021 The Author(s)",

year = "2021",

month = sep,

day = "2",

doi = "10.1016/j.stem.2021.04.001",

language = "אנגלית",

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pages = "1549--1565.e12",

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Bayerl, J, Ayyash, M, Shani, T, Manor, YS, 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, JH 2021, 'Principles of signaling pathway modulation for enhancing human naive pluripotency induction', Cell Stem Cell, vol. 28, no. 9, pp. 1549-1565.e12. https://doi.org/10.1016/j.stem.2021.04.001

TY - JOUR

T1 - Principles of signaling pathway modulation for enhancing human naive pluripotency induction

AU - Bayerl, Jonathan

AU - Ayyash, Muneef

AU - Shani, Tom

AU - Manor, Yair Shlomo

AU - Gafni, Ohad

AU - Massarwa, Rada

AU - Kalma, Yael

AU - Aguilera-Castrejon, Alejandro

AU - Zerbib, Mirie

AU - Amir, Hadar

AU - Sheban, Daoud

AU - Geula, Shay

AU - Mor, Nofar

AU - Weinberger, Leehee

AU - Naveh Tassa, Segev

AU - Krupalnik, Vladislav

AU - Oldak, Bernardo

AU - Livnat, Nir

AU - Tarazi, Shadi

AU - Tawil, Shadi

AU - Wildschutz, Emilie

AU - Ashouokhi, Shahd

AU - Lasman, Lior

AU - Rotter, Varda

AU - Hanna, Suhair

AU - Ben-Yosef, Dalit

AU - Novershtern, Noa

AU - Viukov, Sergey

AU - Hanna, Jacob H.

PY - 2021/9/2

Y1 - 2021/9/2

N2 - Isolating human MEK/ERK signaling-independent pluripotent stem cells (PSCs) with naive pluripotency characteristics while maintaining differentiation competence and (epi)genetic integrity remains challenging. Here, we engineer reporter systems that allow the screening for defined conditions that induce molecular and functional features of human naive pluripotency. Synergistic inhibition of WNT/β-CATENIN, protein kinase C (PKC), and SRC signaling consolidates the induction of teratoma-competent naive human PSCs, with the capacity to differentiate into trophoblast stem cells (TSCs) and extraembryonic naive endodermal (nEND) cells in vitro. Divergent signaling and transcriptional requirements for boosting naive pluripotency were found between mouse and human. P53 depletion in naive hPSCs increased their contribution to mouse-human cross-species chimeric embryos upon priming and differentiation. Finally, MEK/ERK inhibition can be substituted with the inhibition of NOTCH/RBPj, which induces alternative naive-like hPSCs with a diminished risk for deleterious global DNA hypomethylation. Our findings set a framework for defining the signaling foundations of human naive pluripotency.

AB - Isolating human MEK/ERK signaling-independent pluripotent stem cells (PSCs) with naive pluripotency characteristics while maintaining differentiation competence and (epi)genetic integrity remains challenging. Here, we engineer reporter systems that allow the screening for defined conditions that induce molecular and functional features of human naive pluripotency. Synergistic inhibition of WNT/β-CATENIN, protein kinase C (PKC), and SRC signaling consolidates the induction of teratoma-competent naive human PSCs, with the capacity to differentiate into trophoblast stem cells (TSCs) and extraembryonic naive endodermal (nEND) cells in vitro. Divergent signaling and transcriptional requirements for boosting naive pluripotency were found between mouse and human. P53 depletion in naive hPSCs increased their contribution to mouse-human cross-species chimeric embryos upon priming and differentiation. Finally, MEK/ERK inhibition can be substituted with the inhibition of NOTCH/RBPj, which induces alternative naive-like hPSCs with a diminished risk for deleterious global DNA hypomethylation. Our findings set a framework for defining the signaling foundations of human naive pluripotency.

KW - cross-species chimerisim

KW - embryonic stem cells

KW - extra-embryonic stem cells

KW - iPSC

KW - naive pluripotency

KW - reprogramming

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U2 - 10.1016/j.stem.2021.04.001

DO - 10.1016/j.stem.2021.04.001

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C2 - 33915080

AN - SCOPUS:85108881928

SN - 1934-5909

VL - 28

SP - 1549-1565.e12

JO - Cell Stem Cell

JF - Cell Stem Cell

IS - 9

ER -

Principles of signaling pathway modulation for enhancing human naive pluripotency induction

Abstract

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Keywords

UN SDGs

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