Genomic Analysis of Tumor Context Vulnerabilities in Human Metastatic Melanoma

描述（由申请人提供）：尽管经过数十年的研究，转移性恶性黑色素瘤仍然是一种不治之症。只有一小部分诊断为转移性疾病的患者对化疗表现出临床反应。治愈的人更少。转移性疾病患者的五年生存率为5- 18%。因此，本研究的重点是确定黑色素瘤内的遗传变化，通过功能基因组筛选和体内验证证实，这提供了一个可以在晚期疾病中利用的遗传脆弱性的背景。具体而言，申请人建议进行高通量功能性RNA干扰（RNAi）筛选，以系统地鉴定介导黑色素瘤细胞敏感性的基因（具体目标1）。随后，将通过确认siRNA介导的体外基因敲减、进行高含量的机制分析以阐明这些基因介导体外黑素瘤细胞敏感性的机制以及评价与选择剂组合体外抑制这些基因的有效性来验证所鉴定的那些靶点（特定目标2）。最后，申请方将评价经验证的遗传靶点是否可以添加到针对黑色素瘤异种移植模型的体内分子信息联合治疗中（具体目标3）。申请人假设，这种基于功能的基因组方法将鉴定与肿瘤进展相关的遗传畸变的背景，这些遗传畸变对于使黑色素瘤细胞对化疗更具抗性至关重要。这些经过验证的基因可以促进这些肿瘤背景脆弱性的组合靶向，并可能与临床开发直接相关。 公共卫生相关性：患有晚期转移性黑色素瘤的患者的长期预后是暗淡的，五年生存率为5- 18%。这项拟议的研究旨在确定转移性黑色素瘤细胞获得的遗传脆弱性，这些遗传脆弱性可能被用来提高对这种疾病的临床干预的有效性。

DESCRIPTION (provided by applicant): Despite decades of research, metastatic malignant melanoma remains an incurable disease. Only a small percentage of patients diagnosed with metastatic disease exhibit a clinical response to chemotherapy. Fewer still are cured. The five-year survival rate for those with metastatic disease ranges from 5-18%. Therefore, the focus of this study is to identify those genetic changes within melanoma, confirmed through functional genomic screening and in vivo validation, which provide a context of genetic vulnerability that can be exploited in advanced disease. Specifically, the applicant is proposing to perform high-throughput functional RNA interference (RNAi) screen to systematically identify genes that mediate melanoma cell sensitivity (Specific Aim 1). Subsequently, those targets identified will be validated (Specific Aim 2) by confirming siRNA-mediated gene knockdown in vitro, performing high-content mechanistic assays to elucidate the mechanism by which these genes mediate melanoma cell sensitivity in vitro, and evaluating the effectiveness of inhibiting these genes in vitro in combination with the selecting agent. Finally, the applicant will evaluate whether validated genetic targets can add to molecularly-informed combination therapies in vivo against xenograft models of melanoma (Specific Aim 3). The applicant hypothesizes that this functionally-based genomic approach will identify the context of genetic aberrations associated with tumor progression that are critical to rendering melanoma cells more resistant to chemotherapy. These validated genes could facilitate the combined targeting of these tumor-context vulnerabilities and could be of direct relevance to clinical exploitation. PUBLIC HEALTH RELEVANCE: The long-term prognosis for those afflicted with advanced stage metastatic melanoma is bleak, with a five-year survival rate ranging from 5-18%. The proposed research aims to identify genetic vulnerabilities acquired by metastatic melanoma cells that may be exploited to improve the effectiveness of clinical intervention in this disease.