Project 1 - High Throughtput Drug Screening and Correlations with Mutational Status in Myeloma Cell Lines and Patient Samples

项目1摘要/摘要 尽管最近在治疗方面取得了进展，但大多数多发性骨髓瘤(MM)患者并未治愈，而是 患有一种慢性复发，但最终致命的疾病。两个挑战立即变得明显起来。多数 当务之急是需要为未能通过现有有效药物类别的患者找到替代疗法。第二，是 首先了解患者可能会产生抵抗力的原因，并寻求方法学、剂量策略和 可以预防或克服抗药性复发的新药物组合。 我们提出了一种创新的策略，即对500名初级患者样本进行直接药物筛查 解决这两个大问题的化学基因组学审问。我们的假设是一个直接的 对个别患者进行药物分析将提高应答率，降低不必要的毒性，减少药物使用 通过为每个患者确定FDA批准的最有效的药物组合来降低成本。这 战略还将为本提议的目的提供一个样本和临床数据集的数据库 用来探索药物敏感性和耐药性的基因组或临床相关性。 我们的目标将通过在广泛的细胞系基础上成功追求三个具体目标来实现 和初级患者样本数据。首先，我们将测量79种MM疗法的体外敏感性，包括 500例原发性骨髓瘤患者标本中CTEP化合物的检测第二，我们将进行组合筛选， 寻求IMIDS和蛋白酶体抑制剂作为基础化合物与其他活性MM的协同组合 可以在动物模型(如人类CRBN)中测试的治疗方法，如溴域抑制剂 第三，使用我们的M3P突变小组，我们将进行化学基因组学询问，以 利用前处理细胞和存活细胞检测药物敏感性和耐药性的特定相关性 初级病人的机器人屏幕。这些研究将确定特定基因突变或 对多发性骨髓瘤治疗中使用的最有效药物或药物组合具有耐药性的细胞亚群将被共享 在整个计划中进行进一步的表观遗传和转录分析，并获得双向反馈 来自项目2和项目3。 这一战略对目前的U54应用至关重要，它将提供一个包含500个多发性骨髓瘤样本的数据库 以及链接的临床数据集，它们共同构建了药物敏感性和临床表型的马赛克 这项计划拨款的所有要素都可以用来探索药物敏感性和临床相关性的基因组或临床相关性 抵抗。

PROJECT 1 SUMMARY/ABSTRACT Despite recent advances in therapy the majority of multiple myeloma (MM) patients are not cured but rather suffer from a chronic relapsing, yet ultimately fatal disease. Two challenges immediately become evident. Most urgent is the need to find alternative therapies for patients who fail existing potent drug classes. Second, is to understand why patients may be resistant in the first place and to seek methodologies, dosing strategies and new drug combinations which can prevent or overcome drug-resistant relapse. We propose an innovative strategy of “direct to drug” screening of 500 primary patient samples with chemogenomic interrogation which addresses both of these two big questions. Our hypothesis is that a direct to drug analysis of individual patients will improve response rates, lower unnecessary toxicity and reduce drug costs through identification of the most effective combinations of FDA approved drugs for each patient. This strategy will also, for the purposes of this proposal, provide a database of samples and clinical data sets from which to explore genomic or clinical correlates of drug sensitivity and resistance. Our goal will be attained through the successful pursuit of three specific aims, building upon extensive cell line and primary patient sample data. First, we will measure the in vitro sensitivity of 79 MM therapeutics including CTEP compounds in 500 primary myeloma patient samples. Second, we will conduct combination screens to seek synergistic combinations of IMiDs and proteasome inhibitors as base compounds with other active MM therapeutics such as bromodomain inhibitors which can be tested in animal models such as the human CRBN mouse in project 2. Third, using our M3P mutation panels, we will conduct a chemogenomic interrogation to examine specific correlates of drug sensitivity and resistance utilizing pre-treatment and surviving cells from primary patient robotic screens. These studies will determine the frequency of specific genetic mutation or cellular subsets resistant to the most active drugs or drug combinations used in MM therapy and will be shared throughout the program for further epigenetic and transcriptional analysis and bi-directional feedback derived from both Projects 2 and 3. Critical to the current U54 application this strategy will provide a database of 500 multiple myeloma samples and linked clinical data sets which together build a mosaic of drug sensitivity and clinical phenotype from which all elements of this program grant can exploit to explore genomic or clinical correlates of drug sensitivity and resistance.