Regulation of Bcl-2 dependence in multiple myeloma

项目总结/摘要 多发性骨髓瘤是浆细胞的一种不可治愈的恶性肿瘤， 迫切需要各种办法。目前的治疗目标是浆细胞生物学，这被认为是所有人共同的 骨髓瘤然而，对这些治疗的反应是高度可变的，这表明在这些治疗中存在潜在的异质性。 生物学最近发现的一个具有重要治疗意义的异质性来源包括 依赖于Bcl-2蛋白家族，包括Bcl-2、Bcl-xL和Mcl-1。这些蛋白质调节细胞 存活率，并且是正在研究用于骨髓瘤的一类新药的靶点。虽然两者都正常 浆细胞和骨髓瘤通常依赖Mcl-1存活，Mcl-1是骨髓瘤的一个亚群，具有B细胞样 表型是Bcl-2依赖性的并且对维奈托克（一种Bcl-2特异性抑制剂）敏感。这种B细胞表型是 值得注意的是，因为B细胞也是Bcl-2依赖性的，并且B细胞恶性肿瘤如CLL对Bcl-2高度敏感。 维奈托克本提案的目的是检验以下假设：在骨髓瘤的一个亚群中，B细胞 转录和信号网络仍然活跃，并有助于Bcl-2依赖和维奈托克 敏感性以及对常规浆细胞定向治疗的抗性。拟议的研究将确定 骨髓瘤中Bcl-2家族依赖性的机制和鉴定能够选择骨髓瘤的生物标志物 最有可能对Bcl-2抑制剂如维奈托克有反应的患者。目标1将描述 BATF 3活性是一种AP-1转录因子，可增加Bcl-2依赖性并使骨髓瘤对 venetoclax，使用CRISPR介导的BATF 3过表达的细胞的基因组和表观基因组分析，或 击倒。目的2将直接探讨Bcl-2依赖和维奈托克内在耐药的起源 在骨髓瘤中，使用CRISPR筛选靶向在维奈托克敏感性和耐药性中差异表达的基因， 抗性细胞最后，Aim 3将在离体检测的骨髓瘤患者中使用最先进的质谱细胞术 样本开发一种新的生物标志物，可用于未来的精确医学试验，以预测反应 Bcl-2、Bcl-xL和Mcl-1的抑制剂。这些研究将由Vikas Gupta博士进行，他的长期研究 职业目标是成为一个独立资助的医生，科学家研究的分子发病机制， 多发性骨髓瘤他正在寻求NIH K 08职业发展奖，以获得必要的额外技能 为了独立在一个多学科专家小组作为导师和顾问的帮助下，他将 接受先进研究技术的实践培训，如靶向CRISPR筛选，RNA测序， 生物信息学、质谱仪以及指导手稿和资助写作、临床试验 发展，实验室管理和领导技能。完成这里提出的工作将奠定 为R 01未来应用研究克服Bcl-2抗性机制的方法奠定基础 家族抑制剂

PROJECT SUMMARY/ABSTRACT Multiple myeloma is an incurable malignancy of plasma cells for which new treatments and precision guided approaches are acutely needed. Current treatments target the plasma cell biology thought to be common to all myeloma. However, response to these treatments is highly variable pointing to an underlying heterogeneity in biology. One recently identified source of heterogeneity with important therapeutic implications involves dependence on the Bcl-2 family of proteins, which includes Bcl-2, Bcl-xL, and Mcl-1. These proteins regulate cell survival and are the targets of a new class of drugs being studied for use in myeloma. Although both normal plasma cells and myeloma are typically dependent on Mcl-1 for survival, a subset of myeloma with a B cell-like phenotype is Bcl-2 dependent and sensitive to venetoclax, a Bcl-2 specific inhibitor. This B cell phenotype is notable because B cells are also Bcl-2 dependent and B cell malignancies such as CLL are highly sensitive to venetoclax. The objective of this proposal is to test the hypothesis that in a subset of myeloma, B cell transcriptional and signaling networks remain active and contribute to Bcl-2 dependence and venetoclax sensitivity as well as resistance to conventional plasma cell directed therapy. The studies proposed will define the mechanisms of Bcl-2 family dependence in myeloma and identify biomarkers capable of selecting myeloma patients most likely to respond to Bcl-2 inhibitors such as venetoclax. Aim 1 will characterize the mechanisms of BATF3 activity, an AP-1 transcription factor that increases Bcl-2 dependence and sensitizes myeloma to venetoclax, using genomic and epigenomic analyses on cells with CRISPR mediated BATF3 overexpression or knockdown. Aim 2 will directly interrogate the origins of Bcl-2 dependence and intrinsic resistance to venetoclax in myeloma using CRISPR screens targeting genes that are differentially expressed in venetoclax sensitive and resistant cells. Finally, Aim 3 will utilize state of the art mass cytometry on ex vivo tested myeloma patient samples to develop a novel biomarker that could be used for future precision medicine trials to predict response to inhibitors of Bcl-2, Bcl-xL, and Mcl-1. These studies will be conducted by Dr. Vikas Gupta, whose long-term career goal is to become an independently funded physician-scientist studying the molecular pathogenesis of multiple myeloma. He is seeking an NIH K08 career development award to acquire the additional skills necessary for independence. With the help of a multi-disciplinary group of experts serving as mentors and advisors, he will receive hands on training in advanced research techniques such as targeted CRISPR screens, RNA sequencing, bioinformatics, and mass cytometry as well as mentoring in manuscript and grant writing, clinical trial development, laboratory management, and leadership skills. Completion of the work proposed here will lay the foundation for future R01 applications studying approaches to overcome mechanisms of resistance to Bcl-2 family inhibitors.