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.

项目摘要 转化生长因子β（TGFβ）是一种分泌的蛋白质，也是TGFβsu-的第一个成员 要描述的配体的完美。但是，控制TGFβ的分子机制 在其矛盾的生长抑制和上皮 - 间质转变（EMT）之间切换 促进功能尚未完全阐明。由于TGFβ在正常情况下引起多效性功能 发育，成人组织稳态和病理生理过程，例如癌症和纤维化， 未来的研究工作至关重要，集中于对 TGFβ功能。为此，该提案的目的是研究细胞和分子 峰值1（富含假昆虫的非典型激酶One）调节TGFβ信号传导的机制 并在疾病进展过程中介导TGFβ诱导的EMT。拟议的工作是 我们最近发表和引人入胜的初步数据，表明（i）峰介导TGFβ诱导的 EMT，迁移，增殖和癌症转移； （ii）峰1位于膜肌动蛋白结构 并根据TGFβ/纤连蛋白刺激调节SRC/GRB2/MAPK信号传导； （iii）抑制 峰1翻译阻断了TGFβ信号传导的病理生理效应。那，中央假设 该提议的ESI是EIF5A驱动的峰1翻译促进了A组装 在膜ITGB3激活的情况下，SRC/GRB2/Peak1复合物使TGFβ诱导的MAPK能够 信号传导，ZEB1上调和EMT。这种方法具有创新性，因为它将采用一个组合 最先进的细胞，分子，生化，显微镜，蛋白质组学和模型生物的影响 阐明新型EIF5A/peak1翻译的作用机理的方法 SRC/GRB2/Peak1/MAPK细胞骨架信号淋巴结作为TGFβ诱导的EMT的新调节剂。 此外，拟议的研究很重要，因为它将解决以下两个主要 TGFβ研究领域内的挑战和需求：特定目标1将表征翻译 TGFβ诱导的EMT的翻译后机制；具体目标2将确定背景 - 依赖性时空动力学，用于TGFβ反应的分子调节剂。集体 从这些机械研究中获得的知识将识别并表征细胞的细节 以及TGFβ失调以引起疾病的分子环境，以及用于块的新方法 产生TGFβ信号的负面后果。