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

摘要 我们提出了一项发现和治疗开发计划，以建立替代剪接作为 Myc基因驱动前列腺癌的治疗靶点。原癌基因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发挥着核心作用。然而抑制Myc的治疗方法 行动尚未到达诊所。作为转录因子，Myc蛋白很难与任何一种小的 分子或免疫疗法。针对Myc表达下游效应子的间接策略 该计划可能会被证明更成功。 我们的策略是针对剪接因子和多个Myc Paralog依赖于驱动前列腺癌。MYC最近被证明可以控制不同的剪接模式 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 可以识别并可能以小分子为靶点的调节蛋白。同时，我们 假设Myc驱动的选择性剪接将产生适合于 免疫疗法的发展。考虑到Myc去调控在人类癌症中的普遍存在，我们预计 我们的结果将对癌症研究界具有广泛的相关性。 我们在加州大学洛杉矶分校组建了一支在生物信息学方面拥有丰富经验的研究团队(YI Xing)、选择性剪接(Douglas Black)和癌细胞生物学和免疫学(Owen Witte)。我们的 Proposal使用Unique将癌症基因组数据的分析与有重点的实验研究相结合 MYC转化的人类前列腺材料。我们将收集有关依赖于Myc的选择性剪接的数据 来自大型数据集(TCGA、GTEx)以及数据集的正常前列腺组织和原发癌 代表先进的疾病状态。总蛋白质组学分析将对我们转化的 确定候选异构体蛋白表达的材料。我们将采用高通量筛查 我们的一个实验室开发了一个平台，用于识别依赖Myc的剪接的遗传或化学调节器 事件。用于免疫疗法开发的候选细胞表面异构体将携带癌症- 适合利用噬菌体展示文库开发抗体的特定外显子-外显子连接。高亲和力，高 将在CAR T细胞中植入特异性抗体以供进一步开发

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