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

项目1总结/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，而是在治疗过程中出现了严重的并发症。 患有一种慢性复发性但最终致命的疾病两个挑战立即变得明显。最 迫切需要为现有有效药物类别失败的患者找到替代疗法。第二， 了解为什么患者可能首先耐药，并寻求方法，给药策略和 新的药物组合，可以防止或克服耐药复发。 我们提出了一个创新的战略“直接药物”筛选的500个主要患者样本， 化学基因组学研究解决了这两个大问题。我们的假设是， 对个体患者进行药物分析将提高反应率，降低不必要的毒性， 通过为每位患者确定FDA批准的药物的最有效组合来降低成本。这 为了本提案的目的，该战略还将提供一个样本和临床数据集数据库， 以探索药物敏感性和耐药性的基因组或临床相关性。 我们的目标将通过成功追求三个具体目标来实现，建立在广泛的细胞系基础上， 和主要患者样本数据。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样本中的CTEP化合物。第二，我们将进行组合筛选， 寻求IMiD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CRBN中测试的治疗剂如布罗莫结构域抑制剂 项目2中的鼠标。第三，使用我们的M3 P突变面板，我们将进行化学基因组学询问， 利用预处理和存活细胞检查药物敏感性和抗性的特定相关性， 主要患者机器人屏幕。这些研究将确定特定基因突变的频率， 对MM治疗中使用的最有效药物或药物组合耐药的细胞亚群，并将共享 在整个程序中进行进一步的表观遗传和转录分析和双向反馈， 项目2和项目3。 对于当前的U 54应用至关重要，该策略将提供500个多发性骨髓瘤样本的数据库 以及链接的临床数据集，这些数据集一起构建了药物敏感性和临床表型的马赛克， 该计划的所有内容都可以用于探索药物敏感性的基因组或临床相关性， 阻力

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.