A High-Throughput Model for Human Melanoma

描述(申请人提供)：黑色素瘤的发病率在过去40年中增加了600%；它是美国年轻人中增长最快的恶性肿瘤，目前是25-29岁女性癌症死亡的主要原因。如果及早发现，这种疾病很容易治疗，然而，一旦疾病转移，它在很大程度上对传统疗法无效，并与高死亡率相关。黑色素瘤发病率的增加，加上晚期疾病患者预后不良，使得我们必须增加对黑色素瘤潜在遗传原因的理解，以便能够开发更有针对性的治疗策略。人类基因组测序和基因表达微阵列技术的发展导致了数百个在人类癌症中表达发生改变的基因的鉴定。虽然关于肿瘤中表达改变的基因已经获得了大量的知识，但对于这些基因中哪些与肿瘤的发生有因果关系，哪些只是疾病的标志物，人们知之甚少。由于与开发新疗法相关的巨大成本，开发更好的临床前模型来验证哪些基因改变可以有效地作为治疗干预的靶点是至关重要的。这项建议通过描述基于RCAS/TVA逆转录病毒载体系统的黑色素瘤小鼠模型的发展来努力满足这一需求。我们已经建立了从DCT启动子开始在黑素细胞中特异性表达TVA逆转录病毒受体的Ink4A/Arf lox/L？x小鼠，并建议利用含有Cre重组酶和NRAS(Q61R)的逆转录病毒载体诱导DCJ-T/A/lnk4a/Arf L*/L*小鼠发生黑色素瘤。我们将使用这个模型系统来识别和验证与黑色素瘤进展和转移有关的新基因。可以通过使用四环素反应逆转录病毒或能够通过RNA干扰减少基因表达的逆转录病毒下调其表达来识别可以作为治疗靶向的基因。我们的长期目标是将从这些研究中获得的知识转化为治疗晚期黑色素瘤的分子靶向治疗的改进，并将该系统用作未来药物疗效研究的临床前模型。关联性。黑色素瘤发病率的上升，特别是在年轻人和中年人中，是一个重大的公共卫生问题。这项研究的长期目标是利用这项技术来识别黑色素瘤细胞生存所需的关键蛋白质，这些蛋白质可能成为开发更有效的治疗晚期黑色素瘤药物的潜在靶点。

DESCRIPTION (provided by applicant): The incidence of melanoma has increased 600 percent over the last four decades; it is the most rapidly increasing malignancy among young people in the United States and is currently the leading cause of cancer death in women aged 25-29. If detected early, the disease is easily treated, however, once the disease has metastasized it is largely refractory to conventional therapies and is associated with a high mortality rate. The increased incidence of melanoma, combined with the poor prognosis of patients with advanced disease, make it imperative that we increase our understanding of the underlying genetic causes of melanoma such that better targeted therapeutic strategies can be developed. The sequencing of the human genome and the development of gene expression microarray technologies have resulted in the identification of hundreds of genes whose expression is modified in human cancers. While a vast amount of knowledge has been gained about genes with altered expression in tumors, relatively little is known about which of these genes are causally associated with tumor development and which represent only markers for the disease. Because of the great costs associated with the development of new therapies, it is essential that better pre-clinical models are developed to validate which genetic alterations can be productively targeted for therapeutic intervention. This proposal strives to fulfill this need by describing the development of a mouse model of melanoma based on the RCAS/TVA retroviral vector system. We have generated Ink4a/Arf lox/l¿x mice that express the retroviral receptor TVA specifically in melanocytes from the dopachrome tautomerase (DCT) promoter and propose to utilize retroviral vectors containing Cre-recombinase and NRas(Q61R) to induce melanoma in DCJ-T\/A/lnk4a/Arf l¿*/l¿* mice. We will use this model system to identify and validate novel genes responsible for melanoma progression and metastasis. Genes that can be targeted therapeutically will be identified by downregulating their expression using a tetracycline-responsive retrovirus or a retrovirus capable of reducing gene expression via RNA interference. Our long-term goals are to translate the knowledge gained from these studies into improvements in molecular targeted therapies for the treatment of advanced melanoma and to use this system as a pre-clinical model for future studies of drug efficacy. Relevance. The increasing incidence of melanoma, in particular among young to middle-aged adults, is a significant public health problem. The long-term goal of this research is to use this technology to identify key proteins required for the survival of melanoma cells that could serve as potential targets for the development of more effective drugs for the treatment of advanced stage melanoma.