系统性研究癌症中RNA剪接异常的病理功能和分子机理

Most human genes undergo alterative splicing, which is mis-regulated at a global scale in human cancers. Despite the wide spread splicing misregulation in cancer cells, their regulatory mechanism and functional consequence on tumorigenesis are largely unclear. We have used bioinformatic methods to analyze the transcriptome of thousand of cancer patients in 12 types of different cancers, and found more than 70 cancer-specific abnormal splicing in multiple tumors and a large number of RNA-binding proteins with common expression changes in cancers. Therefore, we propose to use both computational and experimental approaches to systematically study the regulatory rules and physiological function of abnormal splicing in cancers. We will also determine the function of RNA-binding proteins in controlling cancer-specific splicing, and examine the role of alternative splicing events in key cell signaling pathways including mTOR pathway, Hippo-YAP pathway, MAPK pathway and p53 pathway. We will specifically manipulate cancer-specific splicing events using CRISPR-cas method or engineered splicing factors with designer specificity, and determine how the splicing manipulations can affect tumor cell proliferation, migration and apoptosis. The studies proposed here will provide new insight to the global regulatory rules of alternative splicing in cancer and its .biological function, which will not only improve our understanding of basic principles in gene regulation but also provide new cancer biomarkers or anti-cancer therapeutic targets.

RNA可变剪接失调与肿瘤密切相关，但剪接异常在肿瘤生长扩散中的功能和机制尚不清楚。我们利用生物信息方法分析了12种癌症中几千个病人样本的剪接数据，发现70余种RNA在多类癌症中都有选择性剪接异变，同时发现大量RNA结合蛋白在肿瘤中表达异常。本课题将运用计算和实验结合的方法，系统性研究这些RNA结合蛋白对异变剪接事件的调控，并阐明癌症中RNA剪接的整体规律和病理生理功能。我们将进一步探讨这些异变剪接事件对mTOR、Hippo-YAP、MAPK、P53等癌症细胞关键信号通路的调控作用，以及这些信号通路中剪接异变对肿瘤发生的生物学意义；最后我们将通过CRISPR和人工剪接因子技术来特异性操控这些异常的RNA剪接事件，在整体水平干预肿瘤的发生发展。本课题的研究结果将有助于系统性解析RNA剪接的整体规律及病理生理功能，并加深对基因调控规则的深入理解，为癌症的诊断治疗提供新的分子标志物和潜在靶点。

RNA剪接失调是肿瘤发生的分子标志之一。剪接因子在癌症中的异常表达和活性改变能够调节许多癌症相关基因的异常剪接，参与了细胞增殖、迁移和凋亡过程。这些异常剪接调控可以作为癌症治疗的新靶点。本项目通过确定肿瘤组织中异常的RNA可变剪接模式在肿瘤发生和生长中的作用，以及明确肿瘤组织中异常表达的RNA结合蛋白在RNA选择性剪接和肿瘤发生中的作用，进一步阐明RNA 结合蛋白在肿瘤细胞周期中的调控机制，并通过构建筛选RNA反式作用因子的功能的PUF-ESF系统，来干预肿瘤发生中的RNA剪接模式。本项目的研究结果有助于系统性解析RNA剪接的整体规律及病理生理功能，并加深对基因调控规则的深入理解，为癌症的诊断治疗提供了新的分子标志物和潜在靶点。