Simple SummaryTERT promoter mutations are the most frequent mutations in melanoma, co-occur regularly with BRAF alterations and are associated with a poorer prognosis. Conflicting results have been published on the role of TERT promoter mutations in resistance to targeted therapy in melanoma. Our data suggest that the TERT mRNA level is associated with resistance to BRAF and MEK inhibitors and could therefore be a more reliable marker for prognosis than the promoter mutations. We showed that overexpression of TERT in a V600E-BRAF melanoma cell line drove resistance to BRAF and MEK inhibitors by a mechanism involving the reactivation of the MAPK pathway independently of telomere maintenance. Finally, we established that TERT inhibition is a therapeutic option in V600E-BRAF-mutated melanoma with acquired resistance to BRAF inhibition. Our results demonstrated the diversity of TERT biological activities in melanoma and highlight the therapeutic potential of targeting TERT in these tumors.AbstractBecause BRAF-mutated melanomas are addicted to the Mitogen Activated Protein Kinase (MAPK) pathway they show a high response rate to BRAF and MEK inhibitors. However, the clinical responses to these inhibitors are often short-lived with the rapid onset of resistance to treatment. Deciphering the molecular mechanisms driving resistance has been the subject of intense research. Recent in vitro and clinical data have suggested a link between expression of telomerase and resistance to targeted therapy in melanoma. TERT promoter mutations are the main mechanism for the continuous upregulation of telomerase in melanoma and co-occur frequently with BRAF alterations. To understand how TERT promoter mutations could be associated with resistance to targeted therapy in melanoma, we conducted translational and in vitro studies. In a cohort of V600E-BRAF-mutated melanoma patients, we showed that the TERT promoter mutation status and TERT expression tended to be associated with response to BRAF and MEK inhibitors. We demonstrated that TERT overexpression in BRAF-mutated melanoma cells reduced sensitivity to BRAF and MEK independently of TERT’s telomer maintenance activity. Interestingly, inhibition of TERT reduced growth of BRAF-mutated melanoma including resistant cells. TERT expression in melanoma can therefore be a new biomarker for resistance to MAPK inhibitors as well as a novel therapeutic target.