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.

Harnessing autophagy to overcome mitogen‐activated protein kinase kinase inhibitor‐induced resistance in metastatic melanoma

“…Previous studies indicate that CD271 expression within primary melanomas correlates with a more aggressive tumour phenotype and reduced patient survival, while isolated CD271‐expressing cells from primary melanomas initiate tumour growth in immunocompromised NRG mice at a higher rate than CD271 – cells . Conversely, other studies suggest that, rather than being a marker of distinctive melanoma‐initiating subpopulations, CD271 expression is induced during acquired resistance to BRAF inhibition, DNA‐damaging drugs or ethanol . Collectively, these studies indicate that CD271 is a biomarker of tumour progression and is consistent with the concept that CD271 signalling constitutes a stress‐tolerance mechanism within tumour cells.…”

Role of RAD51AP1 in homologous recombination DNA repair and carcinogenesis

“…RAD51AP1 transcript was shown to be overexpressed in aggressive mantle cell lymphoma compared to indolent small lymphocytic lymphoma [17], in HPV-positive squamous cell carcinoma of the head and neck (3.6-fold) compared to normal oral epithelium [56], in ovarian cancer as a consequence of downregulated microRNA hsa-mir-140-3p [57, 58], and in basal and triple-negative breast cancers compared to normal mammary tissue [53, 59]. Consistent with a potential role for RAD51AP1 in metastatic disease [51, 52], high RAD51AP1 expression was found to correlate with reduced time of survival of breast and ovarian cancer patients [58, 59]. Interestingly, sporadic cases of childhood papillary thyroid cancer (PTC) showed increased RAD51AP1 message, while post-Chernobyl cases of PTC exhibited decreased RAD51AP1 , indicating that the molecular events in PTC associated with low-dose IR exposure differ from that of patient-matched sporadic PTC [60].…”

“…Elevated (> 4-fold) RAD51AP1 transcript was detected in metastatic melanoma compared to metastasis-free melanoma [51, 52]. RAD51AP1 expression was positively linked to expression of the stem cell marker CD271 and to mechanisms of therapy resistance in metastatic melanoma cells [52, 62]. Elevated expression of RAD51AP1 was also observed in a small set of brain metastasis [52].…”

“…RAD51AP1 expression was positively linked to expression of the stem cell marker CD271 and to mechanisms of therapy resistance in metastatic melanoma cells [52, 62]. Elevated expression of RAD51AP1 was also observed in a small set of brain metastasis [52]. Since RAD51AP1 is involved in mitigating replication stress [15, 16, 20, 33], it is tempting to speculate that its enhanced expression in metastatic tumors may stem from the requirement of metastatic cells to limit replication stress.…”

Melanoma types by in vivo reflectance confocal microscopy correlated with protein and molecular genetic alterations: A pilot study

“…Cutaneous melanoma (CM) is a highly aggressive skin cancer, with different clinical, histopathologic and biologic aspects, suggesting a heterogeneous family of diseases rather than a unique entity . CM subtypes identified at histology have been extensively researched and associated with genetic alterations and expressions of biomarkers, which proved to be useful in prognostic evaluations and clinical and therapeutic decision‐making .…”