Identification of small molecule modulators of MITF

描述（由申请人提供）：小眼症相关转录因子（MITF）是黑素细胞存活和分化的谱系限制性重要调节因子。在15-20%的黑色素瘤中，MITF被扩增并且表现为真正的癌基因：这些黑色素瘤对MITF活性上瘾，并且表达或活性的抑制导致细胞周期停滞、细胞凋亡和致瘤性丧失。鉴于其在黑素细胞和黑色素瘤生物学中的重要性，抑制其活性可能为治疗黑色素瘤提供新的治疗方法。本研究的长期目标是了解调节黑素细胞谱系中MITF表达的信号通路。我们的中心假设是下调MITF可能是黑色素瘤的一种治疗策略。我们的目的是通过高通量筛选（HTS）来鉴定MITF的小分子抑制剂，从而更好地了解MITF的生物学功能，并开发用于黑色素瘤治疗的治疗药物。对于该RO 3资助，我们提出了三个具体目标：（1）使用基于细胞的荧光素酶报告基因测定法对MITF活性调节剂进行小分子筛选，以及（2）进行二级和（3）三级筛选，以验证具体目标1中鉴定的活化化合物。尽管超出了本申请的范围，但我们将尝试开发进一步改进活性化合物的结构和功能的策略。在建议的项目结束时，我们将能够确定几个高度活跃和具体的抑制MITF与进一步研究MITF的生物学功能。这些化合物的鉴定也可能导致黑色素瘤的新治疗方法的发展。 公共卫生相关性：识别小眼症相关转录因子（MITF）的调节因子对于了解色素沉着和频繁突变的黑色素瘤癌基因的主要调节因子至关重要。由于MITF的调节可用于黑色素瘤的治疗，我们期望我们的筛选将与NIH的使命相关，并且将对临床医生以及对黑色素瘤的分子发病机制感兴趣的研究人员感兴趣。

DESCRIPTION (provided by applicant): Micropthalmia-associated transcription factor (MITF) is an lineage-restricted, essential regulator of melanocyte survival and differentiation. In 15-20% of melanomas, MITF is amplified and is a behaves as a bone-fide oncogene: these melanomas are addicted to MITF activity, and suppression of expression or activity leads to cell cycle arrest, apoptosis and loss of tumorigenicity. Given its importance in melanocyte and melanoma biology, suppression of its activity may offer a novel therapeutic approach for the treatment of melanoma. The long-term GOAL of this research is to understand the signaling pathways that modulate MITF expression in the melanocyte lineage. Our CENTRAL HYPOTHESIS is that down-regulation of MITF may a therapeutic strategy for melanoma. Our OBJECTIVE is to identify small molecule suppressors of MITF using high throughput screening (HTS), with which to better understand the biological functions of MITF and to develop therapeutic agents for melanoma therapy. For this RO3 grant, we propose three SPECIFIC AIMS: (1) To perform a small molecule screen for modulators of MITF activity using cell-based luciferase reporter assays and (2) To perform secondary and (3) tertiary screens to validate activate compounds identified in Specific Aim 1. Although beyond the scope of this application, we will attempt to develop strategies for further refinement of the structure and function of active compounds. At the conclusion of the proposed project, we will be able to identify several highly active and specific suppressors of MITF with which to further investigate the biological functions of MITF. The identification of these compounds may also lead to development of novel therapeutic approaches for melanoma. PUBLIC HEALTH RELEVANCE: Identifying modulators of the micropthalmia-associated transcription factor (MITF) is critical to understand this master regulator of pigmentation and frequently mutated melanoma oncogene. Because modulation of MITF can be exploited for the treatment of melanoma, we expect that our screen will be relevant to the mission of NIH and will be interesting to both clinicians as well as researchers interested in the molecular pathogenesis of melanoma.