Abstract 2918: Concomitant inhibition of SIRT1 and SIRT3 reduces melanoma growth and metastasis in BRAFV600E/PTENNULL transgenic mice
Authors: Chhabra Gagan , Singh Chandra K. , Ndiaye Mary A. , Iczkowski Kenneth A. , Ahmad Nihal 
Published: September 2020

Metastatic melanoma, one of the most aggressive types of skin cancer, is extremely difficult to treat, with a median overall survival of less than one year. Despite recent advances in targeted- and immune- therapies, the treatment options have either failed to achieve &gt;25% response or the responses have been short-lived with the emergence of resistance. Therefore, newer approaches are needed for the management of this deadly neoplasm. 4′-bromo-resveratrol [5-(2-(4-hydroxyphenyl)vinyl)-1,3-benzenediol; 4′-BR], an analog of resveratrol, is a dual small molecule inhibitor of sirtuins 1 and 3 (SIRTs 1 & 3), which have pro-proliferative functions in melanoma. In a recently published study, we demonstrated that 4′-BR induced apoptosis and caused metabolic reprogramming in human melanoma cells, leading to reduced proliferation and growth of melanoma cells in vitro. In order to validate our in vitro findings to in vivo situations, in this study, we evaluated the therapeutic efficacy of 4′-BR on melanoma growth and metastasis in a genetically engineered transgenic (BRAFV600E/PTENNull) mouse model of melanoma. This mouse model shows melanoma's cardinal features and is an excellent model to test drug efficacy in pre-clinical settings. To induce melanoma tumors, 4-hydroxytamoxifen was applied on shaved backs of 10-week-old mice topically once per day for 3 consecutive days. Highly pigmented tumors appeared 10 days after tamoxifen application. At this point, treatment with 4′-BR (0, 5, 10, 20 and 30 mg/kg b.wt.; intraperitoneal; 3d/week; n=6 mice/group) was started. Body weight, tumor size and volume were measured weekly. At the termination of the study (~17 weeks of age), we observed significant reduction in tumor volume and tumor weight in mice treated with 30 mg/kg 4'-BR with no noticeable adverse effects. Further, we analyzed markers for cell proliferation and survival (Ki67, PCNA, and Survivin), metastasis (Vimentin), oxidative stress (Nrf2 and Keap1) and growth factor signaling (IGF1 and IGFBP5) using immunohistochemistry (IHC) or real-time quantitative PCR (RT-qPCR) in tumors. IHC analysis revealed that 4′-BR treatment markedly decreased Ki67 positive tumor cells. RT-qPCR analysis showed that 4'-BR treatments significantly decreased expression of PCNA, Survivin, and IGF1 (a melanoma promoting growth factor), as well as significantly increased expression of IGFBP5, a tumor suppressor gene. Furthermore, we observed that 4'-BR treatment significantly decreased the ratio of Nrf2/Keap1. In addition, our data demonstrated a significant decrease in lung metastasis and the levels of metastasis marker Vimentin in 4'-BR treatment group. Overall, these data show the ability of 4′-BR to exert anti-proliferative, anti-tumorigenic, and anti-metastatic effects in a human-relevant melanoma mouse model, thus warranting further pre-clinical and clinical investigations in this direction.

Citation Format: Gagan Chhabra, Chandra K. Singh, Mary A. Ndiaye, Kenneth A. Iczkowski, Nihal Ahmad. Concomitant inhibition of SIRT1 and SIRT3 reduces melanoma growth and metastasis in BRAFV600E/PTENNULL transgenic mice [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2918.