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Stem Cell Reports. 2017 Oct 10;9(4):1234-1245. doi: 10.1016/j.stemcr.2017.08.023. Epub 2017 Sep 28.

An RNAi Screen Reveals an Essential Role for HIPK4 in Human Skin Epithelial Differentiation from iPSCs.

Author information

1
Skin Cancer Unit (G300), German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, 69121 Heidelberg, Germany; Department of Dermatology, Venereology and Allergology, University Medical Center Mannheim, Heidelberg University, 68167 Mannheim, Germany. Electronic address: l.larribere@dkfz.de.
2
Skin Cancer Unit (G300), German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, 69121 Heidelberg, Germany; Department of Dermatology, Venereology and Allergology, University Medical Center Mannheim, Heidelberg University, 68167 Mannheim, Germany.
3
Division of Signaling and Functional Genomics, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany; Department of Cell and Molecular Biology, Heidelberg University, 69120 Heidelberg, Germany; Department of Cardiology, Heidelberg University, 69120 Heidelberg, Germany.
4
Genetics of Skin Carcinogenesis, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany.
5
Genetics of Skin Carcinogenesis, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany; IUF-Leibniz Research Institute for Environmental Medicine, 40021 Düsseldorf, Germany.
6
Division of Signaling and Functional Genomics, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany; Department of Cell and Molecular Biology, Heidelberg University, 69120 Heidelberg, Germany.
7
Skin Cancer Unit (G300), German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, 69121 Heidelberg, Germany; Department of Dermatology, Venereology and Allergology, University Medical Center Mannheim, Heidelberg University, 68167 Mannheim, Germany. Electronic address: j.utikal@dkfz.de.

Abstract

Molecular mechanisms responsible for the development of human skin epithelial cells are incompletely understood. As a consequence, the efficiency to establish a pure skin epithelial cell population from human induced pluripotent stem cells (hiPSCs) remains poor. Using an approach including RNAi and high-throughput imaging of early epithelial cells, we identified candidate kinases involved in their differentiation from hiPSCs. Among these, we found HIPK4 to be an important inhibitor of this process. Indeed, its silencing increased the amount of generated skin epithelial precursors at an early time point, increased the amount of generated keratinocytes at a later time point, and improved growth and differentiation of organotypic cultures, allowing for the formation of a denser basal layer and stratification with the expression of several keratins. Our data bring substantial input regarding regulation of human skin epithelial differentiation and for improving differentiation protocols from pluripotent stem cells.

KEYWORDS:

HIPK4; RNAi screen; development; differentiation; epithelium; iPSC; keratinocytes; organotypic culture; skin; stem cell

PMID:
28966120
PMCID:
PMC5639458
DOI:
10.1016/j.stemcr.2017.08.023
[Indexed for MEDLINE]
Free PMC Article

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