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Transient acquisition of pluripotency during somatic cell transdifferentiation with iPSC reprogramming factors

Nature Biotechnology volume 33, pages 769–774 (2015)Cite this article

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Abstract

Somatic cells can be transdifferentiated to other cell types without passing through a pluripotent state by ectopic expression of appropriate transcription factors1,2. Recent reports have proposed an alternative transdifferentiation method in which fibroblasts are directly converted to various mature somatic cell types by brief expression of the induced pluripotent stem cell (iPSC) reprogramming factors Oct4, Sox2, Klf4 and c-Myc (OSKM) followed by cell expansion in media that promote lineage differentiation3,4,5,6. Here we test this method using genetic lineage tracing for expression of endogenous Nanog and Oct4 and for X chromosome reactivation, as these events mark acquisition of pluripotency. We show that the vast majority of reprogrammed cardiomyocytes or neural stem cells obtained from mouse fibroblasts by OSKM-induced 'transdifferentiation' pass through a transient pluripotent state, and that their derivation is molecularly coupled to iPSC formation mechanisms. Our findings underscore the importance of defining trajectories during cell reprogramming by various methods.

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Figure 1: Lineage tracing for endogenous Nanog reactivation during reprogramming.
Figure 2: Frequent and transient acquisition of pluripotency during OSKM-TD.
Figure 3: Molecular and functional evidence for transient acquisition of pluripotency during OSKM-TD.

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Article 01 December 2022

Zixuan Zhao, Xinyi Chen, … Hanry Yu

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Gene Expression Omnibus

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Acknowledgements

J.H.H. is supported by a generous gift from I. and P. Mantoux; the New York Stem Cell Foundation (NYSCF), FAMRI, the Kimmel Innovator Research Award, the ERC (StG-2011-281906), the Leona M. and Harry B. Helmsley Charitable Trust, Moross Cancer Institute, the Israel Science Foundation Regular research program, the ICRF Foundation, Helen and Martin Kimmel Institute for Stem Cell research (HMKISCR), the Benoziyo Endowment fund. J.H.H. and W.G. are supported by an HFSPO research grant. J.H.H. is a New York Stem Cell Foundation - Robertson Investigator. We thank K. Hochedlinger for mutual exchange of results and discussions before publication. We thank Weizmann Institute management for providing critical financial and infrastructural support.

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Authors and Affiliations

  1. The Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel

    Itay Maza, Inbal Caspi, Asaf Zviran, Elad Chomsky, Yoach Rais, Sergey Viukov, Shay Geula, Leehee Weinberger, Vladislav Krupalnik, Suhair Hanna, Mirie Zerbib, Rada Massarwa, Noa Novershtern & Jacob H Hanna

  2. The Department of Gastroenterology, Rambam Health Care Campus & Bruce Rappaport School of Medicine, Technion Institute of Technology, Haifa, Israel

    Itay Maza

  3. Department of Genetics, Stanford University School of Medicine, Stanford, California, USA

    Jason D Buenrostro

  4. The Department of Pediatrics and the Pediatric Immunology Unit,

    Suhair Hanna

  5. , Rambam Health Care Campus & Bruce Rappaport School of Medicine, Technion Institute of Technology, Haifa, Israel

    Suhair Hanna

  6. Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota, USA

    James R Dutton

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Contributions

I.M. and J.H.H. conceived the idea for this project. I.M. designed and conducted experiments. I.M., N.N., R.M and J.H.H. wrote the manuscript with contributions from most other authors. M.Z. conducted microinjections. S.G., I.C., V.K., S.H. assisted in tissue culture and reprogramming experiments. S.V., Y.R. and I.M. constructed and targeted Nanog-CreER construct. W.J.G. and J.D.B. optimized ATAC-seq protocol and assisted A.Z. in analysis. E.C. and L.W. conducted and analyzed whole genome bisulfite sequencing. A.Z. and N.N. conducted and analyzed RNA-seq analysis. J.R.D. generated and provided engineered Oct4-CreER reporter cells. R.M. assisted in and supervised all microscopy imaging and analysis by I.M. and A.Z. presented in this study. N.N. supervised the validity and analysis of all bioinformatics experiments and results in this manuscript. The authors have no competing interests as defined by Nature Publishing Group, or other interests that might be perceived to influence the results and/or discussion reported in this paper.

Corresponding authors

Correspondence to Rada Massarwa, Noa Novershtern or Jacob H Hanna.

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Maza, I., Caspi, I., Zviran, A. et al. Transient acquisition of pluripotency during somatic cell transdifferentiation with iPSC reprogramming factors. Nat Biotechnol 33, 769–774 (2015). https://doi.org/10.1038/nbt.3270

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