Non-Transcriptional Mechanisms of PEAK1 Action During TGFbeta-Induced EMT

TGFbeta 诱导的 EMT 过程中 PEAK1 作用的非转录机制

• 批准号: 9208078
• 负责人: Jonathan Kelber
• 金额: $ 36.25万
• 依托单位: CALIFORNIA STATE UNIVERSITY NORTHRIDGE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-13 至 2020-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9208078
• 关键词: Actins; Address; Adult; Alpha Cell; Animal Model; Apoptosis Promoter; Binding; Biochemical; Biochemistry; Biological Assay; Cell membrane; Cells; Cicatrix; Complement Factor B; Complex; Cytoskeleton; Data; Development; Disease; Disease Progression; Embryonic Development; Epithelial; Fibronectins; Fibrosis; Funding; Future; Gene Proteins; Goals; Growth; Health; Homeostasis; Human; ITGB3 gene; Immunology; In Vitro; Intervention; Knowledge; Lead; Ligands; MAP Kinase Gene; Malignant Neoplasms; Mediating; Membrane; Mesenchymal; Methods; Microscopy; Modeling; Molecular; Neoplasm Metastasis; Non-Malignant; Pathway interactions; Phosphotransferases; Physiological; Physiological Processes; Positioning Attribute; Process; Productivity; Proteins; Proteomics; Publications; Publishing; Regenerative Medicine; Regulation; Research; Research Training; Role; Science; Signal Pathway; Signal Transduction; Structure; System; TYRP1 gene; Testing; Tissues; Transforming Growth Factor beta; Transforming Growth Factors; Translations; Up-Regulation; Work; Wound Healing; combat; cytokine; design; growth factor receptor-bound protein 2; human disease; in vivo; innovation; insight; member; migration; molecular dynamics; neoplastic cell; novel; novel strategies; programs; protein B; public health relevance; response; spatiotemporal; training opportunity; tumor progression

PROJECT SUMMARY Transforming Growth Factor β (TGFβ) is a secreted protein and the first member of the TGFβ su- perfamily of ligands to be described. However, the molecular mechanisms that govern TGFβ’s ability to switch between its paradoxical growth suppressing and epithelial-mesenchymal transition (EMT) promoting functions remain to be fully elucidated. Since TGFβ elicits pleiotropic functions during normal development, adult tissue homeostasis and pathophysiological processes such as cancer and fibrosis, it is essential that future research efforts focus on producing a complete mechanistic understanding of TGFβ function. To this end, the objective of this proposal is to investigate the cellular and molecular mechanisms by which PEAK1 (pseudopodium-enriched atypical kinase one) regulates TGFβ signaling and mediates TGFβ-induced EMT during disease progression. The proposed work is an extension of our recently published and compelling preliminary data showing that (i) PEAK1 mediates TGFβ-induced EMT, migration, proliferation and cancer metastasis; (ii) PEAK1 localizes to membrane actin structures and regulates Src/Grb2/MAPK signaling in response to TGFβ/fibronectin stimulation; and (iii) inhibition of PEAK1 translation blocks the pathophysiological effects of TGFβ signaling. Thus, the central hypoth- esis of this proposal is that eIF5A-driven PEAK1 translation promotes the assembly of a Src/Grb2/PEAK1 complex in the context of membrane ITGB3 activation to enable TGFβ-induced MAPK signaling, ZEB1 upregulation and EMT. The approach is innovative because it will employ a combina- tion of state-of-the-art cellular, molecular, biochemical, microscopy, proteomic and model organism methods to elucidate the mechanisms of action for the novel eIF5A/PEAK1 translation and Src/Grb2/PEAK1/MAPK cytoskeletal signaling nodes as novel regulators of TGFβ-induced EMT. Furthermore, the proposed research is significant because it will address the following two major challenges and needs within the field of TGFβ research: Specific Aim 1 will characterize translational and post-translational mechanisms of TGFβ-induced EMT; and Specific Aim 2 will identify context- dependent spatiotemporal dynamics for molecular regulators of TGFβ responses. The collective knowledge gained from these mechanistic studies will identify and characterize details of the cellular and molecular contexts in which TGFβ is dysregulated to cause disease, and novel methods for block- ing the negative consequences of TGFβ signaling.

项目总结 转化生长因子β(转化生长因子β)是一种分泌型蛋白，是转化生长因子β家族的第一成员。 每个待描述的配体家族。然而，控制转化生长因子βS能力的分子机制 反常生长抑制和上皮-间充质转化(EMT)之间的转换 促进作用还有待于充分阐明。由于转化生长因子β在正常情况下具有多效性 发育、成人组织动态平衡和病理生理过程，如癌症和纤维化， 至关重要的是，未来的研究努力将重点放在从机制上对 转化生长因子β功能。为此，这项建议的目标是研究细胞和分子 富含伪足的非典型激酶1调节转化生长因子β信号转导机制的研究 并在疾病进展过程中介导转化生长因子β诱导的EMT。拟议的工作是对 我们最近发表的令人信服的初步数据表明：(I)PEAK1介导转化生长因子β诱导 EMT、迁移、增殖与肿瘤转移；(Ii)PEAK1定位于膜肌动蛋白结构 并调节src/grb2/MAPK信号通路，以响应转化生长因子β/纤维连接蛋白刺激；和(Iii)抑制 阻断转化生长因子β信号转导的病理生理效应。因此，中心假设是-- 这一建议的实质是，eIF5A驱动的PEAK1翻译促进了 Src/Grb2/PEAK1复合体在膜ITGB3激活背景下激活转化生长因子β诱导的MAPK 信号转导、ZEB1上调和EMT。这种方法是创新的，因为它将使用一种组合- 最新细胞、分子、生化、显微镜、蛋白质组和模式生物的研究 方法阐明新的eIF5A/PEAK1翻译的作用机制和 SrC/Grb2/PEAK1/MAPK细胞骨架信号转导节点作为转化生长因子β诱导内膜转移的新调节因子。 此外，拟议的研究具有重要意义，因为它将解决以下两个主要问题 转化生长因子β研究领域内的挑战和需求：具体目标1将描述翻译 和转化生长因子β诱导的EMT的翻译后机制；以及特定目标2将识别上下文- 转化生长因子β反应的分子调控的独立时空动力学。集体 从这些机制研究中获得的知识将识别和描述细胞的细节 以及转化生长因子β失控致病的分子背景，以及阻断转化生长因子DNA的新方法。 探讨转化生长因子β信号转导的负面后果。