title: The role of the cancer stem cell marker CD271 in DNA damage response and drug resistance of melanoma cells.
abstract: Several lines of evidence have suggested that stemness and acquired resistance to targeted inhibitors or chemotherapeutics are mechanistically linked. Here we observed high cell surface and total levels of nerve growth factor receptor/CD271, a marker of melanoma-initiating cells, in sub-populations of chemoresistant cell lines. CD271 expression was increased in drug-sensitive cells but not resistant cells in response to DNA-damaging chemotherapeutics etoposide, fotemustine and cisplatin. Comparative analysis of melanoma cells engineered to stably express CD271 or a targeting short hairpin RNA by expression profiling provided numerous genes regulated in a CD271-dependent manner. In-depth analysis of CD271-responsive genes uncovered the association of CD271 with regulation of DNA repair components. In addition, gene set enrichment analysis revealed enrichment of CD271-responsive genes in drug-resistant cells, among them DNA repair components. Moreover, our comparative screen identified the fibroblast growth factor 13 (FGF13) as a target of CD271, highly expressed in chemoresistant cells. Further we show that levels of CD271 determine drug response. Knock-down of CD271 in fotemustine-resistant cells decreased expression of FGF13 and at least partly restored sensitivity to fotemustine. Together, we demonstrate that expression of CD271 is responsible for genes associated with DNA repair and drug response. Further, we identified 110 CD271-responsive genes predominantly expressed in melanoma metastases, among them were NEK2, TOP2A and RAD51AP1 as potential drivers of melanoma metastasis. In addition, we provide mechanistic insight in the regulation of CD271 in response to drugs. We found that CD271 is potentially regulated by p53 and in turn is needed for a proper p53-dependent response to DNA-damaging drugs. In summary, we provide for the first time insight in a CD271-associated signaling network connecting CD271 with DNA repair, drug response and metastasis.
authors.affiliation: German Cancer Consortium (DKTK), German Cancer Research Center (DKFZ), Heidelberg, Germany.|Laboratory of Molecular Tumor Pathology, Institute of Pathology, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Redmer
authors.affiliation: Laboratory of Molecular Tumor Pathology, Institute of Pathology, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: I
authors.initials: I
authors.lastname: Walz
authors.affiliation: Laboratory of Molecular Tumor Pathology, Institute of Pathology, Charité-Universitätsmedizin Berlin, Berlin, Germany.|Institute for Theoretical Biology, Humboldt-Universität zu Berlin, Berlin, Germany.
authors.forename: B
authors.initials: B
authors.lastname: Klinger
authors.affiliation: Laboratory of Molecular Tumor Pathology, Institute of Pathology, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Khouja
authors.affiliation: Laboratory of Molecular Tumor Pathology, Institute of Pathology, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Y
authors.initials: Y
authors.lastname: Welte
authors.affiliation: German Cancer Consortium (DKTK), German Cancer Research Center (DKFZ), Heidelberg, Germany.|Laboratory of Molecular Tumor Pathology, Institute of Pathology, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Schäfer
authors.affiliation: Laboratory of Molecular Tumor Pathology, Institute of Pathology, Charité-Universitätsmedizin Berlin, Berlin, Germany.|CPO-Cellular Phenomics and Oncology Berlin-Buch GmbH, Berlin, Germany.
authors.forename: C
authors.identifier: http://orcid.org/0000-0001-9494-5977
authors.initials: C
authors.lastname: Regenbrecht