Article | Published:

Direct reprogramming of fibroblasts into renal tubular epithelial cells by defined transcription factors

Nature Cell Biology volume 18, pages 12691280 (2016) | Download Citation

Abstract

Direct reprogramming by forced expression of transcription factors can convert one cell type into another. Thus, desired cell types can be generated bypassing pluripotency. However, direct reprogramming towards renal cells remains an unmet challenge. Here, we identify renal cell fate-inducing factors on the basis of their tissue specificity and evolutionarily conserved expression, and demonstrate that combined expression of Emx2, Hnf1b, Hnf4a and Pax8 converts mouse and human fibroblasts into induced renal tubular epithelial cells (iRECs). iRECs exhibit epithelial features, a global gene expression profile resembling their native counterparts, functional properties of differentiated renal tubule cells and sensitivity to nephrotoxic substances. Furthermore, iRECs integrate into kidney organoids and form tubules in decellularized kidneys. Our approach demonstrates that reprogramming factors can be identified by targeted in silico analysis. Renal tubular epithelial cells generated ex vivo by forced expression of transcription factors may facilitate disease modelling, drug and nephrotoxicity testing, and regenerative approaches.

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Acknowledgements

We thank A. Heer, A. Sammarco and B. Joch for excellent technical assistance; the staff of the SFB1140 project Z01 and R. Nitschke (Life Imaging Centre (LIC), Albert-Ludwigs-University Freiburg) for help with confocal microscopy; D. Pfeifer for performing microarrays; M. Follo for FACS analysis; O. Wessely (Cleveland, USA), P. Vize (Calgary, Canada), N. Ueno (National Institute for Basic Biology, Japan) and D. Trono (Lausanne, Switzerland) for generously providing plasmids; M. Köttgen for thoughtful comments on the manuscript. This work was supported by the Emmy Noether Programme to S.S.L. (LI1817/2-1) and S.J.A. (AR732/1-1), Projects B07 of the collaborative research initiative KIDGEM (SFB 1140) to S.S.L. and S.J.A. and project A03 of SFB 850 to S.J.A. by the German Research Foundation (DFG), BIOSS Centre of Biological Signalling Studies, and the Excellence Initiative of the German Research Foundation (GSC-4, Spemann Graduate School) to S.J.A.

Author information

Affiliations

  1. Department of Medicine, Renal Division, Medical Center—University of Freiburg, Faculty of Medicine, University of Freiburg, Hugstetter Straße 55, 79106 Freiburg, Germany

    • Michael M. Kaminski
    • , Jelena Tosic
    • , Catena Kresbach
    • , Hannes Engel
    • , Jonas Klockenbusch
    • , Anna-Lena Müller
    • , Roman Pichler
    • , Florian Grahammer
    • , Oliver Kretz
    • , Tobias B. Huber
    • , Gerd Walz
    • , Sebastian J. Arnold
    •  & Soeren S. Lienkamp
  2. Institute of Experimental and Clinical Pharmacology and Toxicology, University of Freiburg, Faculty of Medicine, Albertstraße 25, 79104 Freiburg, Germany

    • Jelena Tosic
    •  & Sebastian J. Arnold
  3. Spemann Graduate School of Biology and Medicine (SGBM), University of Freiburg, Albertstraße 19a, 79104 Freiburg, Germany

    • Jelena Tosic
  4. Faculty of Biology, University of Freiburg, Schänzlestraße 1, 79104 Freiburg, Germany

    • Jelena Tosic
  5. Department of Neuroanatomy, University of Freiburg, Albertstraße 17, 79104 Freiburg, Germany

    • Oliver Kretz
  6. BIOSS Centre of Biological Signalling Studies, University of Freiburg, Schänzlestraße 18, 79104 Freiburg, Germany

    • Tobias B. Huber
    • , Gerd Walz
    • , Sebastian J. Arnold
    •  & Soeren S. Lienkamp

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Contributions

M.M.K., J.T., C.K., H.E., J.K., A.-L.M., R.P., F.G., O.K. and S.S.L. performed experiments. M.M.K., J.T., S.J.A. and S.S.L. planned and analysed experiments. G.W. and T.B.H. helped with data analysis and during final stages of the manuscript. M.M.K., J.T., S.J.A. and S.S.L. wrote and edited the manuscript. S.S.L. conceived the study.

Competing interests

The authors declare that the following competing financial interest exists: patent applications are pending (EPA 16152655.3 and EP16185290.0).

Corresponding authors

Correspondence to Sebastian J. Arnold or Soeren S. Lienkamp.

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Videos

  1. 1.

    Dome formation of iRECs.

    Confocal 3D-stack of iREC dome formation, as depicted in Fig. 3c. iRECs express membrane GFP (green), the nuclei are stained with Hoechst (blue).

  2. 2.

    Integration of iRECs into kidney organoids.

    Confocal 3D-stack of reaggregated kidney organoid as depicted in Fig. 7c. iRECs express membrane GFP (green), LTL staining for renal tubular cells is depicted in magenta.

  3. 3.

    iRECs form tubular structures in decellularized kidneys.

    Confocal 3D-stack of tubule forming iRECs guided by the extracellular matrix of decellularized kidneys as depicted in Fig. 7f. iRECs express membrane GFP (green), the nuclei are stained with Hoechst (blue).

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DOI

https://doi.org/10.1038/ncb3437

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