Molecular Signatures of Melanoma: Predicting Response to Therapy & Targeting

过去孢子资助期的研究已经确定转录因子MITF是一种新的黑色素瘤 癌基因，代表了一个有希望的治疗靶点。我们在这里提出了一种新的化学品 黑色素瘤抗MITF治疗药物开发的基因组学方法。这种方法是基于 基于基因表达的高通量对基因表达特征的调制研究 筛选(高卢布实验室开发的GE-HTSO方法。人们的预期是，由于这一点 经过努力，将发现具有高临床翻译潜力的化合物。为了实现这些目标， 该项目已被组织成3个具体目标。在目标1中，我们将定义一个基因表达签名 功能获得实验、功能丧失实验相结合的MITF激活 (shRNA介导)，以及黑色素瘤中MITF相关基因表达模式的计算分析 病人样本。然后，我们将MITF激活签名转换为Luminex微珠检测(高达100 转录本)，适用于高通量、低成本的化学筛选。同时，我们将评估 常规收集福尔马林固定石蜡包埋黑色素瘤MITF信号检测的可行性 MITF标志可作为未来的诊断生物标志物和药效学的活检 抗MITF治疗反应的标志物。在目标2中，我们将在高吞吐量下使用MITF签名 筛选以鉴定能够调制MITF特征的小分子化合物，并通过推断， 能够调节MITF活性的。将特别强调对所有~2,000名已获批准的FDA进行筛选 药物，因为这些药物可以最快地推进到临床前和临床试验。我们的 其他GE-HTS筛查的经验是，FDA批准的化合物可能具有意想不到的活性 在这个屏幕上被发现。在目标3中，我们将验证屏幕上出现的点击，并确定 在黑色素瘤转化试验中具有最大效力、最大生物活性的化合物，以及 因此，将优先考虑未来可能的临床开发的化合物。

Work over the past SPORE funding period has established the transcription factor MITF as a new melanoma oncogene, and one that represents a promising therapeutic target. We propose here a novel chemical genomic approach toward the development of anti-MITF therapeutics for melanoma. The approach is based on the modulation of gene expression signatures using the Gene Expression-Based High Throughput Screening (GE-HTSO method developed in the Golub laboratory. The expectation is that as a result of this effort, compounds with high potential for clinical translation will be identified. To accomplish these goals, the project has been organized into 3 Specific Aims. In Aim 1, we will define a gene expression signature of MITF activation through a combination of gain-of-function experiments, loss-of-function experiments (shRNA-mediated), and computational analysis of MITF-associated gene expression patterns in melanoma patient samples. We will then convert that MITF activation signature to a Luminex bead assay (of up to 100 transcripts) that is suitable for high throughput, low cost chemical screening. In parallel, we will evaluate the feasibility of measuring the MITF signature in routinely collected formalin-fixed paraffin-embedded melanoma biopsies such that the MITF signature could serve as a future diagnostic biomarker and pharmacodynamic marker of anti-MITF therapeutic response. In Aim 2, we will use the MITF signature in a high throughtput screen to identify small molecule compounds capable of modulating the MITF signature, and by inference, capable of modulating MITF activity. Special emphasis will be placed on screening all ~ 2,000 FDAapproved drugs because these can be most rapidly advanced to preclinical and clinical testing. Our experience with other GE-HTS screens is that unexpected activities of FDA-approved compounds are likely to be discovered in this screen. In Aim 3, we will validate the hits that emerge from the screen, and identify those compounds with the greatest potency, most biological activity in melanoma transformation assays, and as such will prioritize compounds for possible future clinical development.