Functional identification of CLL drug response and resistance

项目总结： 尽管在过去十年中慢性淋巴细胞性白血病的治疗取得了令人印象深刻的进展，但复发的肿瘤 往往对已知的治疗方法产生抵抗力仍然是一个问题。认识抗药性，开发新的 要将慢性淋巴细胞性白血病转变为一种可治愈的疾病，治疗策略是必要的。除了几个值得注意的例外， 对化疗的反应通常通过细胞凋亡发生，这是细胞程序性死亡的一种形式。我们的目标是 确定使CLL细胞对凋亡敏感的药物，并确定将 防止复发。虽然常规细胞毒性的化学筛选是满足这一要求的一个有吸引力的策略 目标，这种方法由于不能可靠地培养超过48小时的CLL细胞而受到阻碍。 活着。这最终反映了我们在体外化学干扰CLL的能力方面的技术差距，以及 测量功能和临床相关的表型。在这里，我们将使用一种新型的化学筛选 一种称为动态BH3分析(DBP)的方法，该方法能够对化学成分进行功能测量 诱导细胞凋亡变化，只需要6-24小时的体外培养，从而最大限度地 分子保真度和肿瘤细胞活力。使用2000多种药物的专家组，我们不仅可以识别 使CLL细胞对凋亡敏感的新分子，但我们也将确定化学凋亡 个别患者在复发时失去或获得的敏感度。仅能唯一地使人过敏的药物 复发的肿瘤为联合化疗提供了令人兴奋的机会。终于明白了 治疗前和治疗后样本中的凋亡脆弱性的机制，我们将关联我们的 用项目1和项目2中的大分子数据集测量AT的功能性凋亡反应 至少104例慢性淋巴细胞性白血病患者。总而言之，在慢性淋巴细胞性白血病中成功识别出促凋亡药物将不会 这只会促进我们对CLL的分子理解，但可能会带来真正新颖的治疗方案。

Project Summary: Despite impressive therapeutic advances in the treatment of CLL over the past decade, relapsed tumors that are often resistant to known therapies remain a problem. Understanding resistance and developing new treatment strategies will be necessary to turn CLL into a curable disease. With few notable exceptions, responses to chemotherapy typically occur through apoptosis, a form of programmed cell death. Our goal is to identify drugs that sensitize CLL cells to apoptosis, and to identify combinations of drugs that would prevent relapse. While chemical screening of conventional cytotoxicity is an attractive strategy to meet this goal, such an approach is impeded by the inability to reliably culture CLL cells for longer than 48 hours ex vivo. This ultimately reflects a technological gap in our ability to chemically perturb CLL ex vivo, and measure functional and clinically relevant phenotypes. Here, we will use a novel chemical screening approach called dynamic BH3 profiling (DBP), which enables functional measurements of chemically induced apoptotic changes, and requires only 6-24 hours of ex vivo culture, thereby maximizing molecular fidelity and tumor cell viability. Using a panel of over 2000 drugs, not only will we identify novel molecules that sensitize CLL cells for apoptosis, but we will also determine chemical apoptotic sensitivities that are lost or gained on relapse for individual patients. Drugs that uniquely sensitize only relapsed tumors present exciting opportunities for combination chemotherapy. Finally to understand mechanisms of apoptotic vulnerabilities in pre- and post-treatment samples, we will correlate our measurements of functional apoptotic responses with the large molecular datasets in Project 1 and 2 for at least 104 CLL patients. In sum, the successful identification of apoptotic sensitizing drugs in CLL, will not only advance our molecular understanding of CLL, but could result in truly novel therapeutic options.