Proteomic investigation of alternative splicing in the control of cell differentiation

选择性剪接控制细胞分化的蛋白质组学研究

• 批准号: RGPIN-2017-06159
• 负责人: Lu, Yu
• 金额: $ 1.89万
• 依托单位: McMaster University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=710820
• 关键词: Proteomic; investigation; alternative; splicing; control

Alternative splicing plays an important role in generating proteome diversity in eukaryotes, as primary messengers (pre-mRNA) turn into multiple mature messengers by inclusion and exclusion of specific sets of protein-coding exons during the process. Alternative splicing is required for cell differentiation in a wide range of biological processes. Despite its importance, the precise mechanisms controlling alternative splicing during cell differentiation remain poorly understood. Proteomics is a powerful tool to study global protein expression, modification, and interaction in many mechanistic biological studies. To date there has been limited use of proteomics to study alternative splicing, mainly due to the lack of proper methodology. Development and application of proteomic methods to study the global splicing program and the upstream splicing regulators during cell fate changes will be essential to further our understanding of the alternative splicing mechanisms that regulate cell differentiation. My proposed research program seeks to develop and apply proteomic methods to delineate a general framework that mechanistically links signal transduction, splicing factors, and pre-mRNA substrates in controlling cell differentiation. We will use human pluripotent stem cells (PSC) as our model systems of cell differentiation, as they can be conveniently expanded in culture and are able to differentiate into most downstream cell types. We will pursue three specific aims within this proposed NSERC Discovery Grant program. First, we will develop proteomic method to identify upstream splicing regulators of key alternative splicing events that are crucial in directing PSC fate control. Second, we will develop proteomic methods to globally investigate protein splice variants in human PSC before and after differentiation initiation. Last, we will use biochemical and genetic approaches to understand the functions of splice variants and their upstream regulators in human PSC self-renewal and differentiation initiation. Through these studies, we will generate new knowledge about the alternative splicing regulatory mechanisms in eukaryotic cells. These results will also improve our understanding of stem cell and progenitor cell self-renewal and differentiation mechanisms. The proteomic methods developed in this research program can be applied to detect alternative splicing events and upstream regulators that may represent potential therapeutic targets in human diseases such as cancer and neurodegenerative diseases. Furthermore, this research program will allow my laboratory to develop talented young scientists who are crucial for Canada's sustainable economic growth fueled by continuous science and technology development.

选择性剪接在真核生物蛋白质组多样性的产生中起着重要作用，在这一过程中，初级信使（pre-mRNA）通过包含和排除特定的蛋白质编码外显子转变为多个成熟信使。在广泛的生物过程中，细胞分化需要选择性剪接。尽管它很重要，但在细胞分化过程中控制选择性剪接的确切机制仍然知之甚少。