OCT4 induces long-lived dedifferentiated kidney progenitors poised to redifferentiate in 3D kidney spheroids

Dorit Omer; Osnat Cohen Zontag; Yehudit Gnatek; Orit Harari-Steinberg; Oren Pleniceanu; Michael Namestnikov; Ayelet Hashahar Cohen; Malka Nissim-Rafinia; Gal Tam; Tomer Kalisky; Eran Meshorer; Benjamin Dekel

doi:10.1016/j.omtm.2023.04.005

OCT4 induces long-lived dedifferentiated kidney progenitors poised to redifferentiate in 3D kidney spheroids

Dorit Omer, Osnat Cohen Zontag, Yehudit Gnatek, Orit Harari-Steinberg, Oren Pleniceanu, Michael Namestnikov, Ayelet Hashahar Cohen, Malka Nissim-Rafinia, Gal Tam, Tomer Kalisky, Eran Meshorer, Benjamin Dekel

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

Abstract

Upscaling of kidney epithelial cells is crucial for renal regenerative medicine. Nonetheless, the adult kidney lacks a distinct stem cell hierarchy, limiting the ability to long-term propagate clonal populations of primary cells that retain renal identity. Toward this goal, we tested the paradigm of shifting the balance between differentiation and stemness in the kidney by introducing a single pluripotency factor, OCT4. Here we show that ectopic expression of OCT4 in human adult kidney epithelial cells (hKEpC) induces the cells to dedifferentiate, stably proliferate, and clonally emerge over many generations. Control hKEpC dedifferentiate, assume fibroblastic morphology, and completely lose clonogenic capacity. Analysis of gene expression and histone methylation patterns revealed that OCT4 represses the HNF1B gene module, which is critical for kidney epithelial differentiation, and concomitantly activates stemness-related pathways. OCT4-hKEpC can be long-term expanded in the dedifferentiated state that is primed for renal differentiation. Thus, when expanded OCT4-hKEpC are grown as kidney spheroids (OCT4-kSPH), they reactivate the HNF1B gene signature, redifferentiate, and efficiently generate renal structures in vivo. Hence, changes occurring in the cellular state of hKEpC following OCT4 induction, long-term propagation, and 3D aggregation afford rapid scale-up technology of primary renal tissue-forming cells.

Original language	English
Pages (from-to)	329-346
Number of pages	18
Journal	Molecular Therapy Methods and Clinical Development
Volume	29
DOIs	https://doi.org/10.1016/j.omtm.2023.04.005
State	Published - 8 Jun 2023

Bibliographical note

Publisher Copyright:
© 2023 The Authors

Funding

We wish to thank Prof. Zohar Dotan from the Department of Urology, Sheba Medical Center, Tel Hashomer, for providing human kidney tissue samples. The work is supported by ISF (Israel Science Foundation) grants 910/11 and 2071/17, the Israel Ministry of Industry “NOFAR” program, Wolfoson Clore Mayer, and the Sagol Center for Regenerative Medicine, School of Medicine, Tel Aviv University (B.D.); an NIH-NIDDK Diabetic Complications Pilot & Feasibility grant (DK076169), the NephroTools FP7 Marie Curie Initial Training Network (project 289754), and The Lisa and David Pulver Family Foundation (B.D.). E.M. is the Arthur Gutterman Family Chair for Stem Cell Research. This work was partially supported by a generous gift from Arthur Gutterman to E.M. and B.D. D.O. conducted and designed the experiments and wrote the paper, O.H.-S. conducted and designed the experiments, O.C.Z. performed bioinformation analysis, O.P. wrote the paper, Y.G. performed experiments and analysis, M.N. performed experiments and analysis, A.-H.C. performed experiments and analysis, M.N.-R. performed experiments and analysis, G.T. performed experiments and analysis, T.K. designed experiments and analysis, E.M. designed experiments and wrote the paper, and B.D. conducted and designed the experiments and wrote the paper. The authors declare no competing interests. We wish to thank Prof. Zohar Dotan from the Department of Urology, Sheba Medical Center, Tel Hashomer, for providing human kidney tissue samples. The work is supported by ISF (Israel Science Foundation) grants 910/11 and 2071/17 , the Israel Ministry of Industry “NOFAR” program, Wolfoson Clore Mayer , and the Sagol Center for Regenerative Medicine , School of Medicine , Tel Aviv University (B.D.); an NIH - NIDDK Diabetic Complications Pilot & Feasibility grant ( DK076169 ), the NephroTools FP7 Marie Curie Initial Training Network (project 289754 ), and The Lisa and David Pulver Family Foundation (B.D.). E.M. is the Arthur Gutterman Family Chair for Stem Cell Research. This work was partially supported by a generous gift from Arthur Gutterman to E.M. and B.D.

Funders	Funder number
Israel Ministry of Industry
Lisa and David Pulver Family Foundation
NIH-NIDDK
Sagol Center for Regenerative Medicine , School of Medicine , Tel Aviv University
Sagol Center for Regenerative Medicine, School of Medicine, Tel Aviv University
National Institutes of Health
National Institute of Diabetes and Digestive and Kidney Diseases	DK076169
Seventh Framework Programme	289754
Israel Science Foundation	2071/17, 910/11

Keywords

CKD
cell therapy
chronic kidney disease
dedifferentiation
kidney organoid
kidney progenitor
kidney regeneration
kidney spheroid
long-term expansion
redifferentiation
reprogramming
tubuloid

UN SDGs

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

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10.1016/j.omtm.2023.04.005

Cite this

Omer, D., Zontag, O. C., Gnatek, Y., Harari-Steinberg, O., Pleniceanu, O., Namestnikov, M., Cohen, A. H., Nissim-Rafinia, M., Tam, G., Kalisky, T., Meshorer, E., & Dekel, B. (2023). OCT4 induces long-lived dedifferentiated kidney progenitors poised to redifferentiate in 3D kidney spheroids. Molecular Therapy Methods and Clinical Development, 29, 329-346. https://doi.org/10.1016/j.omtm.2023.04.005

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abstract = "Upscaling of kidney epithelial cells is crucial for renal regenerative medicine. Nonetheless, the adult kidney lacks a distinct stem cell hierarchy, limiting the ability to long-term propagate clonal populations of primary cells that retain renal identity. Toward this goal, we tested the paradigm of shifting the balance between differentiation and stemness in the kidney by introducing a single pluripotency factor, OCT4. Here we show that ectopic expression of OCT4 in human adult kidney epithelial cells (hKEpC) induces the cells to dedifferentiate, stably proliferate, and clonally emerge over many generations. Control hKEpC dedifferentiate, assume fibroblastic morphology, and completely lose clonogenic capacity. Analysis of gene expression and histone methylation patterns revealed that OCT4 represses the HNF1B gene module, which is critical for kidney epithelial differentiation, and concomitantly activates stemness-related pathways. OCT4-hKEpC can be long-term expanded in the dedifferentiated state that is primed for renal differentiation. Thus, when expanded OCT4-hKEpC are grown as kidney spheroids (OCT4-kSPH), they reactivate the HNF1B gene signature, redifferentiate, and efficiently generate renal structures in vivo. Hence, changes occurring in the cellular state of hKEpC following OCT4 induction, long-term propagation, and 3D aggregation afford rapid scale-up technology of primary renal tissue-forming cells.",

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Omer, D, Zontag, OC, Gnatek, Y, Harari-Steinberg, O, Pleniceanu, O, Namestnikov, M, Cohen, AH, Nissim-Rafinia, M, Tam, G, Kalisky, T, Meshorer, E & Dekel, B 2023, 'OCT4 induces long-lived dedifferentiated kidney progenitors poised to redifferentiate in 3D kidney spheroids', Molecular Therapy Methods and Clinical Development, vol. 29, pp. 329-346. https://doi.org/10.1016/j.omtm.2023.04.005

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T1 - OCT4 induces long-lived dedifferentiated kidney progenitors poised to redifferentiate in 3D kidney spheroids

AU - Omer, Dorit

AU - Zontag, Osnat Cohen

AU - Gnatek, Yehudit

AU - Harari-Steinberg, Orit

AU - Pleniceanu, Oren

AU - Namestnikov, Michael

AU - Cohen, Ayelet Hashahar

AU - Nissim-Rafinia, Malka

AU - Tam, Gal

AU - Kalisky, Tomer

AU - Meshorer, Eran

AU - Dekel, Benjamin

PY - 2023/6/8

Y1 - 2023/6/8

N2 - Upscaling of kidney epithelial cells is crucial for renal regenerative medicine. Nonetheless, the adult kidney lacks a distinct stem cell hierarchy, limiting the ability to long-term propagate clonal populations of primary cells that retain renal identity. Toward this goal, we tested the paradigm of shifting the balance between differentiation and stemness in the kidney by introducing a single pluripotency factor, OCT4. Here we show that ectopic expression of OCT4 in human adult kidney epithelial cells (hKEpC) induces the cells to dedifferentiate, stably proliferate, and clonally emerge over many generations. Control hKEpC dedifferentiate, assume fibroblastic morphology, and completely lose clonogenic capacity. Analysis of gene expression and histone methylation patterns revealed that OCT4 represses the HNF1B gene module, which is critical for kidney epithelial differentiation, and concomitantly activates stemness-related pathways. OCT4-hKEpC can be long-term expanded in the dedifferentiated state that is primed for renal differentiation. Thus, when expanded OCT4-hKEpC are grown as kidney spheroids (OCT4-kSPH), they reactivate the HNF1B gene signature, redifferentiate, and efficiently generate renal structures in vivo. Hence, changes occurring in the cellular state of hKEpC following OCT4 induction, long-term propagation, and 3D aggregation afford rapid scale-up technology of primary renal tissue-forming cells.

AB - Upscaling of kidney epithelial cells is crucial for renal regenerative medicine. Nonetheless, the adult kidney lacks a distinct stem cell hierarchy, limiting the ability to long-term propagate clonal populations of primary cells that retain renal identity. Toward this goal, we tested the paradigm of shifting the balance between differentiation and stemness in the kidney by introducing a single pluripotency factor, OCT4. Here we show that ectopic expression of OCT4 in human adult kidney epithelial cells (hKEpC) induces the cells to dedifferentiate, stably proliferate, and clonally emerge over many generations. Control hKEpC dedifferentiate, assume fibroblastic morphology, and completely lose clonogenic capacity. Analysis of gene expression and histone methylation patterns revealed that OCT4 represses the HNF1B gene module, which is critical for kidney epithelial differentiation, and concomitantly activates stemness-related pathways. OCT4-hKEpC can be long-term expanded in the dedifferentiated state that is primed for renal differentiation. Thus, when expanded OCT4-hKEpC are grown as kidney spheroids (OCT4-kSPH), they reactivate the HNF1B gene signature, redifferentiate, and efficiently generate renal structures in vivo. Hence, changes occurring in the cellular state of hKEpC following OCT4 induction, long-term propagation, and 3D aggregation afford rapid scale-up technology of primary renal tissue-forming cells.

KW - CKD

KW - cell therapy

KW - chronic kidney disease

KW - dedifferentiation

KW - kidney organoid

KW - kidney progenitor

KW - kidney regeneration

KW - kidney spheroid

KW - long-term expansion

KW - redifferentiation

KW - reprogramming

KW - tubuloid

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JF - Molecular Therapy Methods and Clinical Development

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OCT4 induces long-lived dedifferentiated kidney progenitors poised to redifferentiate in 3D kidney spheroids

Abstract

Bibliographical note

Funding

Keywords

UN SDGs

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