Functional identification of drug response and resistance in Richter's Syndrome

摘要（30行，广泛，长期） 随着更有效的CLL疗法的出现，更优先考虑确定更好的CLL治疗。 里希特综合征（RS），因为这已成为最重要的未满足的医疗需求，以解决 CLL患者。该项目的主要目标是利用功能性精准医疗技术， 了解里希特细胞的生物脆弱性，并确定新的药理干预措施， 增加这些细胞中的凋亡信号。我们的首要任务是直接研究RS患者的活样本 与生物标本中心合作获得的。我们试图找出最有前途的组合， 随后在其他机制资助的RS患者的早期临床试验中进行探索。这份工作 精准医疗的关键是将正确的病人与正确的药物相匹配。基因组测序的策略 取得了一些成功，如用EGFR抑制剂治疗EGFR突变型肺癌;然而，迄今为止， RS还没有屈服于识别有针对性的遗传异常，这将是项目的重点 1.在项目3中，我们提出了一种替代方法，我们称之为功能精准医学。而不是 依靠静态组学数据，我们提出了干扰细胞功能的方法，使我们能够识别活性药物。 这一策略的核心是BH 3分析，这是一种将活细胞中的线粒体暴露于 合成BH 3肽并测量线粒体透化。从这个分析中，我们可以了解哪些抗- 细胞存活所依赖的凋亡蛋白，无论是BCL-2、BCL-XL、MCL-1、其某种组合，或 没有这些蛋白质。该信息具有直接翻译应用，作为临床BH 3模拟抑制剂 目前，这三种蛋白质都已存在。事实上，BH 3谱用于指导BCL-2抑制剂的治疗 Venetoclax用于CLL和AML，Venetoclax现已获得FDA批准的两种适应症。我们 建议使用BH 3谱来鉴定哪种BH 3模拟物在RS中最活跃。BH 3分析可以 还提供了一个概括性的衡量一个细胞是多么接近凋亡的阈值。当与一个 短暂的药物暴露前，动态BH 3谱（DBP）可以识别任何类型的药物， 癌细胞中的凋亡信号，使它们更接近凋亡阈值。我们先前已经 在几种液体和实体肿瘤的情况下证明，这种策略可以准确地识别具有体内 活性，并可以预测患者的临床反应。我们建议使用DBP来识别 活性药物，之后我们将探索它们相互结合以及与适当的BH 3 上面鉴定的模拟物。当我们确定药物的脆弱性，为小组的RS样本进行研究， 项目，我们将比较他们与临床，基因组，转录组学和蛋白质组学注释准备 本P01中的其他项目和核心。我们希望利用这些信息深入了解上游 驱动药物诱导的凋亡信号的信号机制。

Summary (30 lines, broad, long-term) With the advent of more effective therapies in CLL, there is a greater priority on identifying better treatment of Richter's Syndrome (RS), as this has become one of the most important unmet medical needs to address for CLL patients. The main goals of this Project are to utilize functional precision medicine techniques to better understand biological vulnerabilities in Richter's cells and to identify novel pharmacologic interventions to increase apoptotic signaling in these cells. Our priority is on the direct study of viable samples from RS patients obtained in collaboration with the Biospecimens core. We seek to identify the most promising combinations to subsequently explore in early phase clinical trials for patients with RS, funded by other mechanisms. The job of precision medicine is to match the right patient to the right drug. A strategy of genomic sequencing has yielded several successes, such as treating EGFR mutant lung cancer with EGFR inhibitors; however, to date RS has not yielded to the identification of targetable genetic abnormalities, and this will be the focus of Project 1. In Project 3, we propose an alternative approach, which we call functional precision medicine. Rather than rely on static -omic data, we propose to perturb cellular function in ways that allow us to identify active drugs. Central to this strategy is BH3 profiling, a technique in which we expose mitochondria from living cells to synthetic BH3 peptides and measure mitochondrial permeabilization. From this assay we can learn which anti- apoptotic proteins the cell relies on for survival, whether BCL-2, BCL-XL, MCL-1, some combination thereof, or none of these proteins. This information has direct translational application, as clinical BH3 mimetic inhibitors of all three listed proteins now exist. In fact, BH3 profiling was used to direct therapy of the BCL-2 inhibitor venetoclax to CLL and AML, two indications for which venetoclax has now received FDA approval. We propose to use BH3 profiling to identify which BH3 mimetic(s) would be most active in RS. BH3 profiling can also provide a summary measure of how close a cell is to the threshold of apoptosis. When coupled with a brief preceding drug exposure, dynamic BH3 profiling (DBP) can identify drugs from any class that induce apoptotic signaling in cancer cells, moving them closer to the threshold of apoptosis. We have previously demonstrated in several liquid and solid tumor contexts that this strategy accurately identifies drugs with in vivo activity for individual tumors, and can predict clinical response in patients. We propose to use DBP to identify active drugs, after which we will explore their use in combination with each other and with the appropriate BH3 mimetics identified above. As we identify drug vulnerabilities for the panel of RS samples to be studied in this Project, we will compare them with clinical, genomic, transcriptomic, and proteomic annotations prepared by other Projects and Cores in this P01. We hope to use this information to gain insight into the upstream signaling mechanisms that drive drug-induced apoptotic signaling.