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CRISPR ‐assisted receptor deletion reveals distinct roles for ERBB 2 and ERBB 3 in skin keratinocytes
Author(s) -
Dahlhoff Maik,
Gaborit Nadège,
Bultmann Sebastian,
Leonhardt Heinrich,
Yarden Yosef,
Schneider Marlon R.
Publication year - 2017
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/febs.14196
Subject(s) - hacat , erbb3 , keratinocyte , erbb , wound healing , biology , cancer research , cell growth , microbiology and biotechnology , receptor tyrosine kinase , cell culture , signal transduction , immunology , genetics
While the epidermal growth factor receptor ( EGFR ) is an established regulator of skin development and homeostasis, the functions of the related tyrosine kinase receptors ERBB 2 and ERBB 3 in this tissue have only recently been examined. Previously reported, skin‐specific deletion of each of these receptors in mice resulted in similar defects in keratinocyte proliferation and migration, resulting in impaired wound healing and tumorigenesis. Because both ERBB 2 and ERBB 3 are targets for treating an array of cancer types, it is important to examine the consequences of receptor inhibition in human keratinocytes. Here, we employed the CRISPR /Cas9 technology to generate HaCaT cells (an established human keratinocyte cell line) lacking ERBB 2 or ERBB 3. HaCaT clones lacking ERBB 2 or ERBB 3 showed comparable reductions in cell proliferation as assessed by BrdU staining. Apoptosis, in contrast, was reduced in ERBB 3‐deficient HaCaT cells only. Assessment of cell migration using a wound healing (scratch) assay showed that the closure of the wound gaps was completed by 48 h in mock and in ERBB 3 knockout clones. In contrast, this process was considerably delayed in ERBB 2 knockout clones, and a complete closure of the gap in the latter cells did not occur before 72 h. In conclusion, both ERBB 2 and ERBB 3 are essential for normal proliferation of skin keratinocytes, but in contrast to ERBB 3, ERBB 2 is essential for migration of human keratinocytes. These observations might bear significance to patient adverse effects of therapeutic agents targeting ERBB 2 and ERBB 3.