The role and regulation of mTORC2 in cell migration

mTORC2在细胞迁移中的作用和调控

• 批准号: 9900031
• 负责人: Pascale G Charest
• 金额: $ 30.53万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9900031
• 关键词: Actins; Address; Advanced Development; Autoimmune Diseases; Binding; Biochemical Genetics; Breast Cancer Cell; Cardiovascular system; Catalytic Domain; Cell Proliferation; Cell membrane; Cell model; Cells; Complex; Cues; Cytoskeleton; Data; Development; Dictyostelium; Dictyostelium discoideum; Disease; ERBB2 gene; Epidermal Growth Factor; Epidermal Growth Factor Receptor; Epithelial Cells; Experimental Models; F-Actin; FRAP1 gene; Family; Focal Adhesions; Future; Guanosine Triphosphate Phosphohydrolases; Human; Immune; Inflammatory; Knowledge; Literature; MCF10A cells; Malignant Neoplasms; Mediating; Molecular; Movement; Mutate; Neoplasm Metastasis; Oncogenes; PH Domain; Pathologic; Pathway interactions; Phosphotransferases; Physiology; Play; Protein Family; Protein Isoforms; Proteins; Proto-Oncogene Proteins c-akt; Public Health; RAS genes; RIPK3 gene; Regulation; Research; Role; Signal Pathway; Signal Transduction; Testing; Work; base; cancer cell; cancer therapy; cell behavior; cell growth; cell motility; drug development; innovation; insight; interest; mammary epithelium; migration; novel; novel therapeutics; phosphoinositide-3,4,5-triphosphate; ras Proteins; recruit; response; therapeutic development; therapy development; tumor

PROJECT SUMMARY The directed migration of cells is an important cellular behavior key to normal physiology and deregulated in disease, including cancer. Yet, how cells direct their movements in response to migration cues is not understood. The mechanistic Target of Rapamycin Complex 2 (mTORC2) plays an evolutionarily conserved role in regulating the actin cytoskeleton and controlling the migration of cells. mTORC2 is one of two multiprotein signaling complexes formed by the mTOR kinase. mTORC1 is a key regulator of cell growth and proliferation, and its regulation and signaling pathway are well described. Much less is known about mTORC2, but recent research revealed a role for mTORC2 in promoting cancer cell migration and tumor dissemination. The ability to target the mTORC2 pathway in cancer therapy, however, is greatly hampered by a lack of mechanistic insight into how mTORC2 is activated, regulated, and signals to the cell motility machinery. We aim to address this knowledge gap by identifying the key molecular mechanisms that direct mTORC2 activity and function in cell migration. Recently, we discovered that two Ras family GTPases, Ras and Rap1, bind mTOR and the mTORC2 component RIP3/SIN1, respectively, and control mTORC2 activity in the experimental model Dictyostelium discoideum. Importantly, our preliminary data indicate that these mTORC2 regulatory mechanisms are conserved in human cells. Our overall objective is to determine the mechanism and role of Ras and Rap1-mediated regulation of mTORC2 activity and function in cell migration. Our central hypothesis is that Ras and Rap1 independently regulate mTORC2 in response to a migration signal through distinct interactions with components of mTORC2, thereby controlling mTORC2's signaling functions in cell migration and playing a key role in promoting cancer cell migration. We will test our hypothesis in three specific aims. In Aim 1, we will take advantage of the mTOR interaction with the Dictyostelium Ras protein RasC that we have identified to define the mechanism and role of Ras-mediated mTORC2 regulation in cell migration, using Dictyostelium as experimental model. In Aim 2, we will identify the role of Rap1 in regulating mTORC2 function in cell migration, using prototypical human epithelial cells as experimental model. In Aim 3, we will define the mTORC2 pathway controlling breast cancer cell migration, applying our findings from the mechanistic studies performed in Aim 1 and Aim 2 to specifically interrogate the role of Ras- and Rap1- regulated mTORC2 in promoting the migration of breast cancer cells. Altogether, the proposed work will lead to the description of novel molecular mechanisms involved in regulating mTORC2 and cell migration, including that of cancer cells, which will provide innovative opportunities for the development of therapeutic strategies for inhibiting the migration of cancer cells in metastasis. Furthermore, our findings will also advance the development of treatments for other diseases that involve the pathological migration of cells, including cardiovascular, inflammatory and autoimmune disorders, in which mTOR pathways play important roles.

项目概要 细胞的定向迁移是正常生理和解除管制的关键细胞行为 疾病，包括癌症。然而，细胞如何根据迁移线索指导其运动并不清楚。 明白了。雷帕霉素复合物 2 (mTORC2) 的机制靶点在进化上具有保守性 调节肌动蛋白细胞骨架和控制细胞迁移的作用。 mTORC2 是两个之一 由 mTOR 激酶形成的多蛋白信号复合物。 mTORC1 是细胞生长的关键调节因子 增殖及其调控和信号传导途径已有详细描述。人们对 mTORC2 知之甚少， 但最近的研究揭示了 mTORC2 在促进癌细胞迁移和肿瘤扩散方面的作用。 然而，在癌症治疗中靶向 mTORC2 通路的能力因缺乏 深入了解 mTORC2 如何激活、调节并向细胞运动机制发出信号。我们 旨在通过确定指导 mTORC2 活性的关键分子机制来解决这一知识差距 并在细胞迁移中发挥作用。最近，我们发现两个 Ras 家族 GTPases Ras 和 Rap1 结合 mTOR 和 mTORC2 组件 RIP3/SIN1 分别控制 mTORC2 的活性 实验模型盘基网柄菌。重要的是，我们的初步数据表明这些 mTORC2 调节机制在人类细胞中是保守的。我们的总体目标是确定机制 以及 Ras 和 Rap1 介导的 mTORC2 活性和功能在细胞迁移中的调节作用。我们的中央 假设 Ras 和 Rap1 通过独立调节 mTORC2 来响应迁移信号 与 mTORC2 成分的独特相互作用，从而控制 mTORC2 在细胞中的信号传导功能 迁移并在促进癌细胞迁移中发挥关键作用。我们将在三个具体方面检验我们的假设 目标。在目标 1 中，我们将利用 mTOR 与盘基网柄菌 Ras 蛋白 RasC 的相互作用， 我们已经确定了 Ras 介导的 mTORC2 调节在细胞迁移中的机制和作用， 使用盘基网柄菌作为实验模型。在目标 2 中，我们将确定 Rap1 在调节 mTORC2 中的作用 使用原型人上皮细胞作为实验模型，在细胞迁移中发挥作用。在目标 3 中，我们将 应用我们的发现，定义控制乳腺癌细胞迁移的 mTORC2 通路 在目标 1 和目标 2 中进行的机制研究专门询问 Ras- 和 Rap1- 的作用 调节mTORC2促进乳腺癌细胞的迁移。总而言之，拟议的工作将导致 涉及调节 mTORC2 和细胞迁移的新分子机制的描述，包括 癌细胞的研究，这将为开发癌症治疗策略提供创新机会 抑制转移中癌细胞的迁移。此外，我们的研究结果也将推动 开发涉及细胞病理性迁移的其他疾病的治疗方法，包括 心血管、炎症和自身免疫性疾病，其中 mTOR 通路发挥着重要作用。