Article | Published:

Replacing reprogramming factors with antibodies selected from combinatorial antibody libraries

Nature Biotechnology volume 35, pages 960968 (2017) | Download Citation

Abstract

The reprogramming of differentiated cells into induced pluripotent stem cells (iPSCs) is usually achieved by exogenous induction of transcription by factors acting in the nucleus. In contrast, during development, signaling pathways initiated at the membrane induce differentiation. The central idea of this study is to identify antibodies that can catalyze cellular de-differentiation and nuclear reprogramming by acting at the cell surface. We screen a lentiviral library encoding 100 million secreted and membrane-bound single-chain antibodies and identify antibodies that can replace either Sox2 and Myc (c-Myc) or Oct4 during reprogramming of mouse embryonic fibroblasts into iPSCs. We show that one Sox2-replacing antibody antagonizes the membrane-associated protein Basp1, thereby de-repressing nuclear factors WT1, Esrrb and Lin28a (Lin28) independent of Sox2. By manipulating this pathway, we identify three methods to generate iPSCs. Our results establish unbiased selection from autocrine combinatorial antibody libraries as a robust method to discover new biologics and uncover membrane-to-nucleus signaling pathways that regulate pluripotency and cell fate.

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Acknowledgements

We would like to thank J. Gottesfeld, A. Patapoutian, V. Lo Sardo and J. Hazen for assistance and/or helpful discussions; W. Ferguson for technical assistance and K. Spencer for assistance with microscopy. This research was supported by the National Institute on Deafness and other Communication Disorders (DC012592 to K.K.B.), the National Institute of Mental Health (MH102698 to K.K.B.), the California Institute for Regenerative Medicine (RB3-02186 to K.K.B.), the Baxter Family, Norris and Del Webb Foundations (K.K.B.), Las Patronas and the Dorris Neuroscience Center (K.K.B.), a predoctoral fellowship from the California Institute of Regenerative Medicine (J.W.B.) the Andrea Elizabeth Vogt Memorial Award (J.W.B.).

Author information

Author notes

    • Joel W Blanchard
    •  & Jia Xie

    These authors contributed equally to this work.

Affiliations

  1. Department of Molecular and Cellular Neuroscience, Dorris Neuroscience Center, The Scripps Research Institute, La Jolla, California, USA.

    • Joel W Blanchard
    • , Nadja El-Mecharrafie
    • , Simon Gross
    • , Sohyon Lee
    •  & Kristin K Baldwin
  2. Department of Cell and Molecular Biology, The Scripps Research Institute, La Jolla, California, USA.

    • Jia Xie
    •  & Richard A Lerner

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Contributions

Experiments were initially conceived by R.A.L. and K.K.B. Subsequent experimental design was contributed by all authors, and experiments were designed, performed and analyzed by J.W.B. and J.X. Experiments were also performed by S.L., S.G. and N.E.-M. The manuscript was written and revised by J.W.B., J.X. and K.K.B.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Jia Xie or Kristin K Baldwin.

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DOI

https://doi.org/10.1038/nbt.3963