Molecular Signatures of Melanoma: Predicting Response to Therapy & Targeting

在过去的SPORE资助期间的工作已经将转录因子MITF确定为一种新的黑色素瘤 癌基因，一个代表一个有前途的治疗靶点。我们在这里提出一种新的化学物质 基因组方法，以发展抗MITF治疗黑色素瘤。该方法是基于 使用基于基因表达的高吞吐量 筛选（Golub实验室开发的GE-HTSO方法。人们期望，由于这一点， 通过努力，将确定具有高临床转化潜力的化合物。为了实现这些目标， 该项目分为三个具体目标。在目标1中，我们将定义以下基因表达特征： 通过功能获得实验、功能丧失实验的组合激活MITF （shRNA介导的）和计算分析黑素瘤中MITF相关基因表达模式 患者样本。然后，我们将MITF激活特征转换为Luminex珠测定（高达100 转录物），其适用于高通量、低成本的化学筛选。同时，我们将评估 在常规采集的福尔马林固定石蜡包埋黑色素瘤中测量MITF特征的可行性 活检，这样MITF签名可以作为未来的诊断生物标志物和药效学 抗MITF治疗反应的标志物。在目标2中，我们将在高吞吐量中使用MITF签名 筛选以鉴定能够调节MITF特征的小分子化合物，并且通过推断， 能够调节MITF活性。将特别强调筛选所有FDA批准的约2，000个 因为这些药物可以最快地进入临床前和临床试验。我们 根据其他GE-HTS筛选的经验，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，我们将验证从屏幕上出现的命中，并识别 在黑色素瘤转化试验中具有最大效力、最大生物活性的那些化合物，以及 因此将优先考虑用于可能的未来临床开发的化合物。

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.