Direct reprogramming of fibroblasts into renal tubular epithelial cells by defined transcription factors.
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.
Department of Medicine, Renal Division, Medical Center-University of Freiburg, Faculty of Medicine, University of Freiburg, Hugstetter Straße 55, 79106 Freiburg, Germany.
Michael M
MM
Kaminski
Department of Medicine, Renal Division, Medical Center-University of Freiburg, Faculty of Medicine, University of Freiburg, Hugstetter Straße 55, 79106 Freiburg, Germany.|Institute of Experimental and Clinical Pharmacology and Toxicology, University of Freiburg, Faculty of Medicine, Albertstraße 25, 79104 Freiburg, Germany.|Spemann Graduate School of Biology and Medicine (SGBM), University of Freiburg, Albertstraße 19a, 79104 Freiburg, Germany.|Faculty of Biology, University of Freiburg, Schänzlestraße 1, 79104 Freiburg, Germany.
Jelena
J
Tosic
Department of Medicine, Renal Division, Medical Center-University of Freiburg, Faculty of Medicine, University of Freiburg, Hugstetter Straße 55, 79106 Freiburg, Germany.
Catena
C
Kresbach
Department of Medicine, Renal Division, Medical Center-University of Freiburg, Faculty of Medicine, University of Freiburg, Hugstetter Straße 55, 79106 Freiburg, Germany.
Hannes
H
Engel
Department of Medicine, Renal Division, Medical Center-University of Freiburg, Faculty of Medicine, University of Freiburg, Hugstetter Straße 55, 79106 Freiburg, Germany.
Jonas
J
Klockenbusch
Department of Medicine, Renal Division, Medical Center-University of Freiburg, Faculty of Medicine, University of Freiburg, Hugstetter Straße 55, 79106 Freiburg, Germany.
Anna-Lena
AL
Müller
Department of Medicine, Renal Division, Medical Center-University of Freiburg, Faculty of Medicine, University of Freiburg, Hugstetter Straße 55, 79106 Freiburg, Germany.
Roman
R
Pichler
Department of Medicine, Renal Division, Medical Center-University of Freiburg, Faculty of Medicine, University of Freiburg, Hugstetter Straße 55, 79106 Freiburg, Germany.
Florian
F
Grahammer
Department of Medicine, Renal Division, Medical Center-University of Freiburg, Faculty of Medicine, University of Freiburg, Hugstetter Straße 55, 79106 Freiburg, Germany.|Department of Neuroanatomy, University of Freiburg, Albertstraße 17, 79104 Freiburg, Germany.
Oliver
O
Kretz
Department of Medicine, Renal Division, Medical Center-University of Freiburg, Faculty of Medicine, University of Freiburg, Hugstetter Straße 55, 79106 Freiburg, Germany.|BIOSS Centre of Biological Signalling Studies, University of Freiburg, Schänzlestraße 18, 79104 Freiburg, Germany.
Tobias B
TB
Huber
Department of Medicine, Renal Division, Medical Center-University of Freiburg, Faculty of Medicine, University of Freiburg, Hugstetter Straße 55, 79106 Freiburg, Germany.|BIOSS Centre of Biological Signalling Studies, University of Freiburg, Schänzlestraße 18, 79104 Freiburg, Germany.
Gerd
G
Walz
Department of Medicine, Renal Division, Medical Center-University of Freiburg, Faculty of Medicine, University of Freiburg, Hugstetter Straße 55, 79106 Freiburg, Germany.|Institute of Experimental and Clinical Pharmacology and Toxicology, University of Freiburg, Faculty of Medicine, Albertstraße 25, 79104 Freiburg, Germany.|BIOSS Centre of Biological Signalling Studies, University of Freiburg, Schänzlestraße 18, 79104 Freiburg, Germany.
Sebastian J
SJ
Arnold
Department of Medicine, Renal Division, Medical Center-University of Freiburg, Faculty of Medicine, University of Freiburg, Hugstetter Straße 55, 79106 Freiburg, Germany.|BIOSS Centre of Biological Signalling Studies, University of Freiburg, Schänzlestraße 18, 79104 Freiburg, Germany.
Soeren S
SS
Lienkamp
D000818:Animals; D002449:Cell Aggregation; D019070:Cell Lineage; D049109:Cell Proliferation; D048430:Cell Shape; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D016000:Cluster Analysis; D004622:Embryo, Mammalian / Q000166:cytology; D004847:Epithelial Cells / Q000166:cytology* / Q000648:ultrastructure; D005347:Fibroblasts / Q000166:cytology*; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling; D006801:Humans; D007684:Kidney Tubules / Q000166:cytology*; D051379:Mice; D009399:Nephrons / Q000166:cytology / Q000378:metabolism; D009940:Organoids / Q000166:cytology; D014157:Transcription Factors / Q000378:metabolism*; D014981:Xenopus
D014157:Transcription Factors