由于临床持久反应不佳，对免疫治疗有抵抗力的恶性黑色素瘤仍然是最致命的皮肤癌形式。基因毒性或靶向化疗等替代疗法在治疗后会引发各种癌细胞命运，包括细胞死亡和衰老。可以使用衰老药物消除衰老细胞，我们假设有针对性地消除治疗诱导的衰老黑色素瘤细胞可以补充传统疗法和免疫疗法。我们利用一组代表与黑色素瘤相关的不同突变背景的细胞，发现它们响应治疗而产生不同的衰老表型。无论 BRAF 突变谱如何，卡铂-紫杉醇或放射线的基因毒性联合疗法都会引发细胞死亡和衰老的混合反应。 DNA 损伤诱导的衰老黑色素瘤细胞表现出形态变化、残留 DNA 损伤和衰老相关分泌表型 (SASP) 增加。相比之下，Braf 和 Mek 的双重靶向抑制引发了不同的混合细胞命运反应，包括衰老样细胞和持续细胞。虽然存留细胞可以增殖，但衰老样细胞被稳定地抑制，但没有检测到 DNA 损伤和衰老相关的分泌表型。为了评估对 senolytics 的敏感性，我们采用了一种新型的基于实时成像的死亡测定，并观察到 ​​Bcl2/Bcl-XL 抑制剂和胡椒长明能有效促进卡铂-紫杉醇和辐射诱导的衰老黑色素瘤细胞的死亡，而混合持久剂Braf-Mek 抑制产生的细胞和衰老样细胞仍然没有反应。有趣的是，当在衰老背景下使用时，观察到 Bcl2/Bcl-XL 抑制剂和 Braf-Mek 抑制剂之间的直接协同作用。总体而言，我们强调了黑色素瘤衰老状态的不同特征，并提供了可以降低治疗耐药性的环境依赖性衰老治疗的证据，同时还讨论了这种策略在人类黑色素瘤细胞中的局限性。版权所有 © 2024 Tchelougou、Malaquin、Cardin、Desmul、Turcote 和 Rodier 。

Malignant Melanoma that resists immunotherapy remains the deadliest form of skin cancer owing to poor clinically lasting responses. Alternative like genotoxic or targeted chemotherapy trigger various cancer cell fates after treatment including cell death and senescence. Senescent cells can be eliminated using senolytic drugs and we hypothesize that the targeted elimination of therapy-induced senescent melanoma cells could complement both conventional and immunotherapies. We utilized a panel of cells representing diverse mutational background relevant to melanoma and found that they developed distinct senescent phenotypes in response to treatment. A genotoxic combination therapy of carboplatin-paclitaxel or irradiation triggered a mixed response of cell death and senescence, irrespective of BRAF mutation profiles. DNA damage-induced senescent melanoma cells exhibited morphological changes, residual DNA damage, and increased senescence-associated secretory phenotype (SASP). In contrast, dual targeted inhibition of Braf and Mek triggered a different mixed cell fate response including senescent-like and persister cells. While persister cells could reproliferate, senescent-like cells were stably arrested, but without detectable DNA damage and senescence-associated secretory phenotype. To assess the sensitivity to senolytics we employed a novel real-time imaging-based death assay and observed that Bcl2/Bcl-XL inhibitors and piperlongumine were effective in promoting death of carboplatin-paclitaxel and irradiation-induced senescent melanoma cells, while the mixed persister cells and senescent-like cells resulting from Braf-Mek inhibition remained unresponsive. Interestingly, a direct synergy between Bcl2/Bcl-XL inhibitors and Braf-Mek inhibitors was observed when used out of the context of senescence. Overall, we highlight diverse hallmarks of melanoma senescent states and provide evidence of context-dependent senotherapeutics that could reduce treatment resistance while also discussing the limitations of this strategy in human melanoma cells.Copyright © 2024 Tchelougou, Malaquin, Cardin, Desmul, Turcotte and Rodier.