Genomic Analysis of Tumor Context Vulnerabilities in Human Metastatic Melanoma

人类转移性黑色素瘤肿瘤背景脆弱性的基因组分析

基本信息

批准号：
7454966
负责人：
JEFFREY M. TRENT
金额：
$ 45.89万
依托单位：
TRANSLATIONAL GENOMICS RESEARCH INST
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-07-01 至 2012-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7454966
关键词：
Biological Assay Candidate Disease Gene Cell Line Cells Clinical Clinical effectiveness Combined Modality Therapy Data Data Analyses Diagnosis Disease Dose Drug Sensitization Effectiveness Evaluation Event Functional RNA Gene Silencing Gene Targeting Genes Genetic Genetic Predisposition to Disease Genomics Goals Heterogeneity Human Image In Vitro Intervention Mediating Melanoma Cell Metastatic Melanoma Minority Modeling Mutation Patients Pharmaceutical Preparations Polymerase Chain Reaction RNA Interference RNA library Range Research Resistance Role Screening procedure Small Interfering RNA Staging Survival Rate Testing Time Tissue Microarray Validation Western Blotting Xenograft Model base cancer cell chemotherapy functional genomics improved in vivo melanoma outcome forecast protein expression response temozolomide tumor tumor progression

项目摘要

DESCRIPTION (provided by applicant): Despite decades of research, metastatic malignant melanoma remains an incurable disease. Only a minority of patients diagnosed with metastatic melanoma show a clinical response to chemotherapy, while still fewer are cured; the 5-year survival rate for patients with disseminated disease ranges from 5-18%. The focus and underlying goal of this project is to identify specific genetic changes in melanoma, confirmed through both functional genomic screening and in vivo validation, which provide a context of genetic vulnerability that can be exploited in advanced stage melanoma. Specifically, we are proposing to undertake a high-throughput functional RNA interference (RNAi) screen to systematically identify genes that mediate melanoma cell sensitivity (Specific Aim 1). We will subsequently validate the role of these genes (Specific Aim 2) by confirming siRNA-mediated gene knockdown in vitro, characterizing protein expression in human melanomas using tissue microarrays (TMAs), 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. Lastly, we will evaluate whether our validated genetic targets can add to molecularly-informed combination therapies in vivo against xenograft models of melanoma (Specific Aim 3). We hypothesize that this functional-based genomic approach will identify the context of genetic perturbations associated with tumor progression that are critical to rendering melanoma cells more resistant to chemotherapy. We speculate that these validated genes could facilitate the combined targeting of these tumor-context vulnerabilities and could be of direct relevance to clinical exploitation. Relevance: The long-term prognosis for patients diagnosed with advanced stage melanoma is dismal, 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.

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无法治愈的疾病。只有少数被诊断患有转移性黑色素瘤的患者对化疗表现出临床反应，而治愈的患者则更少；患有播散性疾病的患者的 5 年生存率为 5-18%。该项目的重点和根本目标是确定黑色素瘤中的特定遗传变化，并通过功能基因组筛选和体内验证进行确认，从而提供可在晚期黑色素瘤中利用的遗传脆弱性背景。具体来说，我们建议进行高通量功能性 RNA 干扰 (RNAi) 筛选，以系统地识别介导黑色素瘤细胞敏感性的基因（具体目标 1）。随后，我们将通过在体外确认 siRNA 介导的基因敲除、使用组织微阵列 (TMA) 表征人类黑色素瘤中的蛋白质表达、进行高内涵机制分析来阐明这些基因的作用机制，从而验证这些基因的作用（具体目标 2）。基因在体外介导黑色素瘤细胞敏感性，并评估与选择剂组合在体外抑制这些基因的有效性。最后，我们将评估我们经过验证的遗传靶标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疗法中（具体目标 3）。我们假设这种基于功能的基因组方法将识别与肿瘤进展相关的遗传扰动的背景，这对于使黑色素瘤细胞对化疗更具抵抗力至关重要。我们推测这些经过验证的基因可以促进针对这些肿瘤背景脆弱性的联合靶向，并且可能与临床开发直接相关。相关性：诊断为晚期黑色素瘤的患者的长期预后很差，五年生存率为 5-18%。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，这些脆弱性可用于提高该疾病临床干预的有效性。