Investigation of the role of insulin receptor in chromosome stability.

研究胰岛素受体在染色体稳定性中的作用。

• 批准号: 10461138
• 负责人: Eunhee Choi
• 金额: $ 40.5万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-05 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10461138
• 关键词: Affect; Aneuploidy; Binding; Biochemistry; CRISPR/Cas technology; Cell Differentiation process; Cell Proliferation; Cell division; Cell membrane; Cell surface; Cells; Cellular Metabolic Process; Cellular biology; Chromosomal Stability; Chromosome Segregation; Clathrin Adaptors; Clinical Treatment; Complex; Cryoelectron Microscopy; Diabetes Mellitus; Disease; Endocytosis; Ensure; Future; Genes; Genomics; Goals; Growth; Growth Factor; Growth Factor Receptors; Health; Homeostasis; Human; Hyperinsulinism; Immune; Insulin; Insulin Receptor; Insulin-Like-Growth Factor I Receptor; Investigation; Knowledge; Ligands; MAP Kinase Gene; MXI1 gene; Malignant Neoplasms; Metabolic; Metabolism; Mitogens; Mitosis; Mitotic Checkpoint; Mus; Non-Insulin-Dependent Diabetes Mellitus; Nutrient; Organism; Outcome; Pathway interactions; Physiological; Proliferating; Proteins; Proto-Oncogene Proteins c-akt; Receptor Mediated Signal Transduction; Receptor Protein-Tyrosine Kinases; Receptor Signaling; Regulation; Research; Risk; Role; Signal Pathway; Signal Transduction; Surface; T-Cell Lymphoma; Therapeutic; Tissues; Translational Research; cancer cell; cancer immunotherapy; extracellular; genome-wide; loss of function; mouse genetics; novel; receptor function; recruit; spatiotemporal; tumor growth

PROJECT SUMMARY Multicellular organisms develop receptor-mediated signal transduction initiated by extracellular growth factors to proliferate. Insulin has long been known as a growth factor, and hyperinsulinemia can promote and sustain tumor growth. Insulin receptor (IR) localizes to the cell surface plasma membrane both in metabolic tissue cells and in highly proliferative cells such as immune cells and cancer cells. IR activates two downstream signaling pathways, the PI3K-AKT pathway and the MAPK pathway, to regulate cell metabolism, proliferation, and growth. Despite intriguing findings for IR signaling in systemic homeostasis, how the nutrient signaling maintains chromosome stability still remains uncertain. This gap in our knowledge presents a key barrier to our understanding of the function of insulin in cell proliferation and differentiation and its impact on human health, as hyperinsulinemia is associated with various diseases including type 2 diabetes and cancer. The key spindle checkpoint protein MAD2 forms a mitotic checkpoint complex (MCC) and ensures the fidelity of chromosome segregation. Our recent studies showed that MAD2 binds to IR, recruits the clathrin adaptor complex by assembling an MCC-like complex, and promotes IR endocytosis. In our unpublished results, we found that disruption of IR-MAD2 interaction in mice increases aneuploidy in the immune cells and promotes T-cell lymphoma. These results suggest that the spindle checkpoint regulators and IR mutually regulate each other in both mitosis and IR signaling. IR and insulin- like growth factor 1 receptor (IGF1R) are highly homologous receptor tyrosine kinases (RTKs). IGF1R does not bind to MAD2, and its endocytosis mechanism and signaling outcomes are different from that of IR. We recently defined the distinct activation mechanisms of IR and IGF1R, suggesting that ligand-specific induced structural differences might affect the endocytosis and downstream signaling of RTKs. In parallel to our studies on the role of spindle checkpoint proteins in IR signaling, we have also performed unbiased, systematic genome-wide loss of function studies using CRISPR-Cas9. We identified novel genes that increase or decrease the levels of surface IR and IR signaling. Here, we propose to combine approaches in mouse genetics, cell biology, biochemistry, genomics and cryo-EM to determine the function of IR in mitosis, as well as activation mechanisms of IR signaling for cell proliferation vs. metabolism. Our goals over the next five years are to explore the following questions: (1) how does IR ensure accurate chromosome segregation in mitosis, and what is the physiological function for IR in cell division; (2) how does IR selectively activate the PI3K-AKT vs. MAPK signaling branch; and (3) what other factors are required for IR function in cell proliferation and differentiation? Collectively, the proposed research will advance our understanding of the function, regulation, and mechanism(s) of insulin action in physiological cell proliferation and differentiation. Furthermore, our studies will likely serve as a basis for further translational research and future therapeutics as hyperinsulinemia and type 2 diabetes are associated with increased risks for certain cancers and may also be harnessed for cancer immunotherapies.

项目概要 多细胞生物发展由细胞外生长因子启动的受体介导的信号转导 增生。胰岛素长期以来被认为是一种生长因子，高胰岛素血症可以促进和维持肿瘤 生长。胰岛素受体 (IR) 定位于代谢组织细胞和体内的细胞表面质膜。 高度增殖的细胞，例如免疫细胞和癌细胞。 IR 激活两条下游信号通路， PI3K-AKT 通路和 MAPK 通路，调节细胞代谢、增殖和生长。尽管 IR信号在系统稳态中的有趣发现，营养信号如何维持染色体 稳定性仍存在不确定性。我们知识上的这种差距是我们理解这一问题的一个关键障碍。 胰岛素在细胞增殖和分化中的作用及其对人类健康的影响，如高胰岛素血症 与多种疾病有关，包括2型糖尿病和癌症。关键纺锤体检查点蛋白 MAD2 形成有丝分裂检查点复合体 (MCC) 并确保染色体分离的保真度。我们最近的 研究表明，MAD2 与 IR 结合，通过组装 MCC 样复合物来招募网格蛋白接头复合物， 并促进 IR 内吞作用。在我们未发表的结果中，我们发现 IR-MAD2 相互作用的破坏 小鼠免疫细胞的非整倍性增加并促进 T 细胞淋巴瘤。这些结果表明 纺锤体检查点调节因子和 IR 在有丝分裂和 IR 信号传导中相互调节。 IR 和胰岛素- 与生长因子 1 受体 (IGF1R) 一样，都是高度同源的受体酪氨酸激酶 (RTK)。 IGF1R 不 与MAD2结合，其胞吞机制和信号传导结果与IR不同。我们最近 定义了 IR 和 IGF1R 的不同激活机制，表明配体特异性诱导结构 差异可能会影响 RTK 的内吞作用和下游信号传导。在我们对角色的研究的同时 IR 信号传导中纺锤体检查点蛋白的研究，我们还进行了公正的、系统的全基因组丢失 使用 CRISPR-Cas9 进行功能研究。我们发现了增加或减少表面水平的新基因 IR 和 IR 信号传输。在这里，我们建议结合小鼠遗传学、细胞生物学、生物化学、 基因组学和冷冻电镜确定 IR 在有丝分裂中的功能以及 IR 信号传导的激活机制 细胞增殖与代谢。我们未来五年的目标是探讨以下问题：（1） IR如何确保有丝分裂中染色体的准确分离，IR的生理功能是什么 在细胞分裂中； (2) IR如何选择性激活PI3K-AKT vs. MAPK信号分支；以及（3）还有什么其他的 细胞增殖和分化中IR功能需要哪些因子？总的来说，拟议的研究 将增进我们对生理学中胰岛素作用的功能、调节和机制的理解 细胞增殖和分化。此外，我们的研究可能会作为进一步转化的基础 研究和未来的治疗方法，因为高胰岛素血症和 2 型糖尿病与风险增加有关 对于某些癌症，也可用于癌症免疫疗法。