The convergence of senescence and ferroptosis amplifies DNA damage and triggers apoptosis in patient-derived melanoma models

Abstract Therapy-induced senescence (TIS) is a common response to anticancer treatment that enforces durable cell-cycle arrest but can also promote tumor persistence and disease relapse if senescent cells are not efficiently eliminated. The “one–two punch” therapeutic paradigm aims to overcome this limitation by coupling senescence induction with senolytic targeting. In this study, using distinct, clinically relevant BRAF- and NRAS-mutant melanoma PDXs, we identify a novel combination strategy in which a vulnerable cell state is induced and then selectively targeted. Through a high-throughput one–two punch screen, we identified auranofin as a top candidate selectively targeting senescent melanoma cells. Auranofin synergized with multiple senescence-inducing agents, including volasertib, palbociclib, everolimus, and temozolomide, and retained activity across multiple PDX models. In particular, the PLK1 inhibitor volasertib induced robust senescence characterized by G2/M arrest and senescence-associated β-galactosidase expression, while auranofin triggered oxidative stress through thioredoxin reductase inhibition. Sequential volasertib–auranofin treatment resulted in marked accumulation of reactive oxygen species, intracellular iron, and metabolic stress, consistent with activation of ferroptotic pathways. Importantly, ferroptosis cooperated with senescence to amplify DNA damage and genomic instability, as evidenced by increased γH2AX foci, nuclear enlargement, and micronuclei formation, culminating in apoptotic cell death. In vivo, combination therapy significantly suppressed tumor growth and prolonged mice survival without overt toxicity, demonstrating robust therapeutic efficacy in both BRAF- and NRAS-mutant melanoma PDXs. Together, our findings define a mechanistic framework in which volasertib-driven senescence creates a ferroptosis-sensitive state that is therapeutically exploited by auranofin, leading to oxidative stress–mediated DNA damage and tumor cell elimination. This study identifies ferroptosis as a critical vulnerability of senescent cells and provides strong preclinical rationale for repurposing auranofin in combination with senescence-inducing therapies, supporting rapid translational advancement of this strategy for aggressive melanoma.