Regulation of self-renewal of pluripotent stem cells by integrin signaling

整合素信号传导调节多能干细胞的自我更新

• 批准号: 2026049
• 负责人: Luis Villa Diaz
• 金额: $ 55.02万
• 依托单位: Oakland University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2026049&HistoricalAwards=false
• 关键词: Regulation; self; renewal; pluripotent; stem

Stem cells are cells with the unique ability to divide into identical daughter cells or to differentiate into specialized cell types. A better understanding of stem cell biology will advance knowledge of how organs and tissues are formed and maintained during the life of an organism, as stem cells are responsible for those functions. In particular, this project will elucidate the role of three different components of a signaling pathway preventing the differentiation of pluripotent stem cells, which translates into how stem cells are maintained during an individual’s life span. In addition, it will provide training opportunities for two graduate students and multiple undergraduates, who will be directly involved in the research activities of this project. Furthermore, dozens of graduate students will be trained in methods to effectively communicate scientific knowledge to the public.The long-term goal of the principal investigator is to elucidate the molecular signaling mechanisms regulating quiescence, self-renewal, and differentiation of stem cells and their connection with the microenvironment in which they reside. In this project, the investigators will determine the role of specific spliced isoforms of the transmembrane protein integrin alpha6 in the inactivation of its heterodimer partner integrin beta1, which results in signaling that sustains self-renewal of pluripotent stem cells. For this, the investigators will use cell-sorting of stem cell populations containing the specific spliced isoform and their genetic manipulation to determine their molecular regulation. In addition, the molecular function of two protein kinases involved in integrin signaling: Focal Adhesion Kinase (FAK) and Integrin-Linked Kinase (ILK), will be determined. The hypothesis that both kinases play key roles as scaffold proteins promoting the degradation of transcription factors related to cell lineage determination, and therefore sustaining self-renewal of pluripotent stem cells, will be tested. For this, the investigators will verify the complexation of these scaffold proteins with specific ubiquitinase proteins and targeted transcription factors by immunoprecipitation assays, and chemical/genetic manipulations of these interactions. The potential findings from these experiments may have significant implications as they could provide a better understanding of how integrin signaling pathways mediate the interconnection between stem cells and their microenvironment.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

干细胞是具有分裂成相同的子细胞或分化成专门细胞类型的独特能力的细胞。 对干细胞生物学的更好理解将促进对器官和组织在生物体生命期间如何形成和维持的知识，因为干细胞负责这些功能。特别是，该项目将阐明阻止多能干细胞分化的信号通路的三种不同组分的作用，这转化为干细胞在个体生命周期中的维持方式。此外，它将为两名研究生和多名本科生提供培训机会，他们将直接参与本项目的研究活动。此外，还将培养数十名研究生，使他们掌握向公众有效传播科学知识的方法。长期目标是阐明调节干细胞静止、自我更新和分化的分子信号机制及其与微环境的联系。在这个项目中，研究人员将确定跨膜蛋白整合素α 6的特定剪接亚型在其异二聚体伴侣整合素β 1失活中的作用，这导致维持多能干细胞自我更新的信号传导。为此，研究人员将使用含有特定剪接异构体的干细胞群的细胞分选及其遗传操作来确定其分子调控。此外，将确定参与整联蛋白信号传导的两种蛋白激酶的分子功能：粘着斑激酶（FAK）和整联蛋白连接激酶（ILK）。这两种激酶作为支架蛋白发挥关键作用，促进与细胞谱系决定相关的转录因子降解，从而维持多能干细胞的自我更新的假设将被检验。为此，研究人员将通过免疫沉淀试验和这些相互作用的化学/遗传操作来验证这些支架蛋白与特定泛素酶蛋白和靶向转录因子的复合。这些实验的潜在发现可能具有重要意义，因为它们可以更好地了解整合素信号通路如何介导干细胞与其微环境之间的相互联系。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估来支持。