Multi-omic analysis of Myc-driven splicing for prostate cancer therapeutic development

摘要 我们提出了一个发现和治疗开发计划，以建立选择性剪接作为一种治疗方法。 Myc驱动的前列腺癌的治疗靶点。原癌基因的Myc家族，包括c-Myc，N-Myc， 和L-Myc，在这种癌症和许多其他癌症的发病机制中起核心作用。然而，抑制Myc的疗法 行动尚未到达诊所。作为转录因子，Myc蛋白很难被靶向， 分子或免疫治疗剂。靶向Myc表达下游效应子的间接策略 计划可能会更成功。 我们的策略是靶向剪接因子和选择性剪接异构体， 是导致前列腺癌的关键Myc基因最近被证明可以控制选择性剪接模式 对Myc驱动的肿瘤生长至关重要。我们假设这些肿瘤依赖于特定的剪接 可以被识别并可能被小分子靶向的调节蛋白。同时，我们 假设Myc驱动可变剪接将产生适合于 免疫治疗发展。鉴于Myc失调在人类癌症中的普遍存在，我们预计， 我们的研究结果将对癌症研究界具有广泛的意义。 我们在加州大学洛杉矶分校组建了一个具有丰富生物信息学经验的研究团队（Yi Xing）、选择性剪接（道格拉斯布莱克）和癌细胞生物学和免疫学（Owen Witte）。我们 该提案将癌症基因组数据的分析与使用独特的 myc转化的人前列腺材料。我们将收集Myc依赖性可变剪接的数据， 来自大型数据集（TCGA，GTEx）的正常前列腺组织和原发性癌症以及数据集 代表着晚期疾病将对我们的转化蛋白进行总蛋白质组学分析。 用于确认候选同种型的蛋白质表达的材料。我们将采用高通量筛选 在我们的一个实验室开发的平台，用于识别Myc依赖性剪接的遗传或化学调节剂 事件选择用于免疫疗法开发的候选细胞表面同种型将携带癌症- 特异性外显子-外显子连接，适合于用噬菌体展示文库开发抗体。高亲和力，高- 特异性抗体将被构建到CAR T细胞中以进行进一步的开发

ABSTRACT We present a discovery and therapeutics development program to establish alternative splicing as a therapeutic target of Myc-driven prostate cancer. The Myc-family of proto- oncogenes, including c-Myc, N-Myc, and L-Myc, play a central role in the pathogenesis of this and many other cancers. Yet therapies inhibiting Myc action have yet to reach the clinic. As transcription factors, Myc proteins are difficult to target with either small molecules or immunotherapeutics. Indirect strategies that target downstream effectors of the Myc expression program may prove more successful. Our strategy is to target the splicing factors and alternatively spliced isoforms that the multiple Myc paralogs rely on to drive prostate cancer. Myc has been recently shown to control alternative splicing patterns that are crucial to Myc-driven tumor growth. We hypothesize that these tumors rely on specific splicing regulatory proteins that can be identified and potentially targeted with small molecules. In parallel, we hypothesize that Myc-driven alternative splicing will create cancer-specific protein isoforms suitable for immunotherapeutics development. Given the ubiquity of Myc deregulation in human cancer, we anticipate that our results will be of broad relevance to the cancer research community. We have assembled a team of investigators at UCLA with extensive experience in bioinformatics (Yi Xing), alternative splicing (Douglas Black), and cancer cell biology and immunology (Owen Witte). Our proposal integrates the analyses of cancer genomic data with focused experimental research using unique Myc-transformed human prostate materials. We will gather data on Myc-dependent alternative splicing in normal prostate tissues and primary cancers from large datasets (TCGA, GTEx) as well as datasets representing advanced disease states. Total proteomics analysis will be conducted on our transformed materials to confirm protein expression of candidate isoforms. We will employ a high-throughput screening platform developed in one of our labs to identify genetic or chemical modulators of Myc-dependent splicing events. Candidate cell surface isoforms selected for immunotherapeutics development will carry cancer- specific exon-exon junctions suitable for antibody development with phage display libraries. High-affinity, high- specificity antibodies will be built into CAR T-cells for further development