Identifying endosomal Phosphatidylinositol 4-phosphate 5-kinase isoforms regulating growth factor stimulated integrin recycling and migration

鉴定调节生长因子刺激的整合素回收和迁移的内体磷脂酰肌醇 4-磷酸 5-激酶亚型

• 批准号: 10217317
• 负责人: Lorraine C Santy
• 金额: $ 16.05万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10217317
• 关键词: Actins; Adhesions; Affect; Automobile Driving; Binding; Biosensor; Carcinoma; Cell Adhesion; Cell membrane; Cell surface; Cells; Chemicals; Consumption; Core Protein; Cytoskeleton; Development; Dimerization; Disease; Disease Progression; EGF gene; Endosomes; Enzymes; Epithelial; Epithelial Cells; Extracellular Matrix; Fibrosis; Focal Adhesions; Functional disorder; Generations; Genes; Growth Factor; Homeostasis; Hyperactivity; Individual; Integrin Binding; Integrins; Intervention; Knowledge; Lead; Lipid Binding; Lipids; Location; Malignant Epithelial Cell; Membrane; Membrane Proteins; Metastatic to; Monomeric GTP-Binding Proteins; Mutation; Neoplasm Metastasis; Normal tissue morphology; Oncogenic; Pathway interactions; Phosphatidylinositols; Phosphorylation; Phosphotransferases; Play; Process; Production; Protein Isoforms; Protein Kinase; Proteins; RNA Splicing; Recycling; Role; Serum; Side; Signal Pathway; Signal Transduction; Specificity; Spliced Genes; Stromal Cells; Structure; System; Tail; Testing; Tissues; Traction; Variant; Work; base; cancer cell; cancer invasiveness; cell motility; epithelial repair; extracellular; fighting; genetic regulatory protein; inorganic phosphate; knock-down; migration; migration stimulating factor; phosphatidylinositol 4-phosphate; phosphoinositide-3,4,5-triphosphate; recruit; repaired; response; restoration; therapy development; tissue repair; trafficking; tumor microenvironment; tumor progression; tumorigenesis; wound healing

PROJECT SUMMARY Epithelial cells are normally non-motile but can become migratory in response to growth factor signaling. Growth factor stimulated migration plays important roles in development and is an early step in the repair process after tissue damage. During cancer progression however, growth factor signaling can become aberrantly activated through oncogenic mutation or the actions of stromal cells in the tumor microenvironment. Growth factor stimulated migration of carcinoma cells promotes metastasis and further disease progression. A thorough understanding of the signals that initiate migration in response to growth factor signaling is essential for developing interventions to either enhance migration during wound healing or inhibit adhesion and migration of cancer cells. Focal adhesions attach cells to the extracellular matrix and allow the generation of traction forces that cells use to migrate. By necessity, growth factor stimulated migration requires increased assembly of new focal adhesions, which are organized by integrins. Growth factor stimulation leads to the rapid recycling of previously internalized integrins to the plasma membrane. Previous work in the Santy lab demonstrated that cytohesin activity is required for stimulated integrin recycling and that only those cytohesin splice variants that bind phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) can initiate this trafficking process. PI(4,5)P2 is produced locally where needed from PI4P by phosphatidylinositol 4-phosphate 5-kinases (PIP5Ks), implicating PIP5Ks in the stimulated recycling of integrins. Previous work on these understudied kinases has predominately focused on their actions at the plasma membrane during signaling and internalization of membrane proteins. The proposed studies will determine which PIP5K isoforms are required for stimulated integrin recycling and determine where within the endosomal system they produce PI(4,5)P2 to initiate stimulated integrin recycling. A knockdown and rescue strategy will be used to test the impact of PIP5K splice isoforms on integrin recycling. The location where these isoforms act will be determined by localization of both the isoforms and their product, PI(4,5)P2. In an orthogonal approach, chemically induced dimerization will be used to recruit PI(4,5)P2 producing and consuming enzymes to individual endosomal compartments to determine where local production of this lipid is required to stimulate the return of integrins to the cell surface. Understanding the steps that lead to growth factor stimulated integrin recycling will allow development of interventions to modulate this process without affecting growth factor signaling as a whole.

项目摘要 上皮细胞通常是非运动的，但可以响应生长因子信号传导而变化。 生长因子刺激的迁移在发展中起着重要作用，并且是修复的早期一步 组织损伤后的过程。然而，在癌症进展过程中，生长因子信号传导可能会变成 通过致癌突变或肿瘤微环境中基质细胞的作用异常激活。 生长因子刺激癌细胞的迁移促进转移和进一步的疾病进展。一个 对响应生长因子信号启动迁移的信号的透彻理解是必不可少的 为了制定干预措施以增强伤口愈合过程中的迁移或抑制粘附和 癌细胞的迁移。局灶性粘连将细胞连接到细胞外基质上，并允许产生 细胞用来迁移的牵引力。必要时，生长因子刺激的迁移需要增加 由整联蛋白组织的新焦点粘连组装。生长因子刺激导致 先前内部整合素的快速回收到质膜。桑蒂实验室的先前工作 证明刺激整联蛋白回收需要细胞粘着蛋白活性，只有那些细胞粘着蛋白 结合磷脂酰肌醇4,5-双磷酸（PI（4,5）P2）的剪接变体可以启动这种运输过程。 Pi（4,5）P2在局部生产在局部由磷脂酰肌醇4-磷酸5-激酶从PI4P所需的地方产生 （pip5ks），将pip5ks牵涉到整联蛋白的刺激回收中。这些研究的先前工作 激酶在信号传导和 膜蛋白的内在化。拟议的研究将确定需要哪些PIP5K同工型 为了刺激整联蛋白回收并确定内体系统中的何处，它们会产生pi（4,5）p2 启动刺激的整联蛋白回收。敲低和救援策略将用于测试PIP5K的影响 整联蛋白回收的剪接同工型。这些同工型法将通过本地化确定的位置 同工型及其产物Pi（4,5）P2中的两个。在正交方法中，化学诱导的二聚化 将用于招募PI（4,5）P2生产和食用酶 确定需要该脂质的局部产生，以刺激整联蛋白向细胞表面的返回。 了解导致生长因子刺激整联蛋白回收的步骤将允许发展 调节此过程的干预措施而不影响整个生长因子信号传导。