Mechanisms of FGF Receptor Regulation and Signaling

FGF 受体调节和信号转导机制

• 批准号: 8487214
• 负责人: MOOSA MOHAMMADI
• 金额: $ 80.79万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8487214
• 关键词: 1-Phosphatidylinositol 3-Kinase; Achievement; Adopted; Affect; Affinity; Agonist; Animals; Applications Grants; Bile Acids; Binding; Biochemical; Biological; Biological Assay; Biomedical Engineering; Bypass; C-terminal; Calorimetry; Cells; Cholesterol; Chronic Kidney Failure; Colon Carcinoma; Complex; Craniosynostosis; Crystallography; Data; Dependency; Development; Diabetes Mellitus; Disease; Embryonic Development; Endocrine; Event; FGF1 gene; FGF21 gene; FGF7 gene; FGF8 gene; FGF9 gene; FGFR2 gene; FGFR4 gene; Fibroblast Growth Factor; Fibroblast Growth Factor Receptors; Foundations; Funding; Glucose; Goals; Grant; Homeostasis; Human; Human Development; Kidney; Kinetics; Knowledge; Laboratories; Link; Lipids; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Mediation; Mesoderm; Metabolism; Mitogen-Activated Protein Kinase Kinases; Molecular; Molecular Conformation; Molecular Mechanisms of Action; Morphogenesis; Mutation; N-terminal; NGFR gene; Neurodegenerative Disorders; Obesity; Organogenesis; PTB Domain; Pathway interactions; Patients; Pattern; Pattern Formation; Pharmaceutical Preparations; Phosphorylation; Phosphotransferases; Physiology; Play; Protein Engineering; Protein Isoforms; Protein Tyrosine Kinase; Public Health; Publications; Receptor Activation; Receptor Protein-Tyrosine Kinases; Receptor Signaling; Recombinant Proteins; Recruitment Activity; Regulation; Replacement Therapy; Role; Scaffolding Protein; Serum; Signal Transduction; Specificity; Spectrum Analysis; Spermatogenesis; Stem cells; Structure; Surface Plasmon Resonance; Syndrome; Tail; Therapeutic; Time; Tissues; Titrations; Tyrosine; Tyrosine Kinase Domain; Tyrosine Phosphorylation; Vitamin D; base; driving force; drug development; drug discovery; gain of function; gain of function mutation; hearing impairment; human FRS2 protein; human disease; human embryonic stem cell; hypercholesterolemia; inorganic phosphate; insight; novel therapeutics; paracrine; protein expression; public health relevance; receptor; receptor binding; reproductive; response; self-renewal; skeletal; somitogenesis; stem cell differentiation; tissue repair; wasting

DESCRIPTION (provided by applicant): Fibroblast growth factor (FGF) signaling plays pleiotropic roles in mammalian development and metabolism, and disease. The paracrine FGF1, FGF4, FGF7, FGF8, and FGF9 subfamilies play essential roles in spermatogenesis, mesoderm induction, somitogenesis, organogenesis, and pattern formation, whereas the FGF19 subfamily acts in an endocrine fashion to regulate major metabolic processes including glucose, lipid, cholesterol, and bile acid metabolism, and phosphate/vitamin D homeostasis. The diverse activities of FGFs are transmitted by the FGF receptor (FGFR) subfamily of receptor tyrosine kinases (RTKs). Perturbed FGF signaling leads to numerous human diseases, including skeletal, reproductive syndromes, hearing loss, renal phosphate wasting, neurodegenerative disorders, and cancer. Several paracrine FGFs and all the endocrine FGFs are being pursued for drug development. The four specific aims of this competing renewal are: I. Characterize the structural basis by which epithelially-expressed FGF4 and FGF9 subfamilies attain their specificity towards mesenchymally-expressed FGFRc isoforms. II. Elucidate the structural basis by which a/bKlotho co-receptors promote signaling by the endocrine FGFs. III. Dissect the role of A-loop tyrosine phosphorylation in the hyperactivation of FGFR tyrosine kinase by pathogenic gain-of-function mutations. IV. Elucidate the structural basis by which FGFR recruits and phosphorylates FRS2a. Recombinant protein expression and engineering, x-ray crystallography, Surface Plasmon Resonance (SPR) spectroscopy, isothermal titration calorimetry (ITC), steady-state kinetics analysis, and time-resolved mass spectrometry will be used to accomplish the Specific Aims of this proposal. The structural and biophysical/biochemical results obtained will also be validated using cell- and animal-based assays. The data obtained under Aim I should provide molecular insights into the roles of paracrine FGFs in embryonic development and also facilitate the discovery of drugs for tissue repair and bioengineering, promotion of self- renewal and differentiation of human embryonic stem cells for cell-replacement therapy. The data generated under Aim II should enhance our understanding of the role of endocrine FGFs in human metabolism and provide blueprints for drug discovery for major human diseases including diabetes, obesity, hypercholesterolemia, colon cancer, and chronic kidney disease, most of which represent a huge burden on public health. The results of Aim III will enhance our understanding of the mechanism of action of pathogenic mutations in FGFRs and other RTKs as well as the regulation of tyrosine kinase activity of the entire RTK superfamily. Since substrate recruitment and phosphorylation by RTKs is a general event in RTK signaling, the mechanistic insights gained under Aim IV will also be directly applicable to the entire RTK superfamily.

描述（由申请人提供）：成纤维细胞生长因子（FGF）信号在哺乳动物发育和代谢以及疾病中发挥多效性作用。旁分泌 FGF1、FGF4、FGF7、FGF8 和 FGF9 亚家族在精子发生、中胚层诱导、体细胞发生、器官发生和模式形成中发挥重要作用，而 FGF19 亚家族以内分泌方式发挥作用，调节主要代谢过程，包括葡萄糖、脂质、胆固醇和胆汁酸 代谢和磷酸盐/维生素 D 稳态。 FGF 的多种活性由受体酪氨酸激酶 (RTK) 的 FGF 受体 (FGFR) 亚家族传递。 FGF 信号传导紊乱会导致多种人类疾病，包括骨骼、生殖综合征、听力损失、肾磷酸盐消耗、神经退行性疾病和癌症。几种旁分泌型 FGF 和所有内分泌型 FGF 正在用于药物开发。这种竞争性更新的四个具体目标是： I. 表征上皮表达的 FGF4 和 FGF9 亚家族获得对间充质表达的 FGFRc 亚型的特异性的结构基础。二.阐明 a/bKlotho 共受体促进内分泌 FGF 信号传导的结构基础。三．剖析 A 环酪氨酸磷酸化在致病性功能获得性突变引起的 FGFR 酪氨酸激酶过度激活中的作用。四．阐明 FGFR 募集和磷酸化 FRS2a 的结构基础。重组蛋白表达和工程、X射线晶体学、表面等离子共振（SPR）光谱、等温滴定量热法（ITC）、稳态动力学分析和时间分辨质谱将用于实现本提案的具体目标。获得的结构和生物物理/生化结果也将使用基于细胞和动物的测定进行验证。目标 I 下获得的数据应为旁分泌 FGF 在胚胎发育中的作用提供分子见解，并促进组织修复和生物工程药物的发现，促进人类胚胎干细胞的自我更新和分化以用于细胞替代疗法。 Aim II 产生的数据应该增强我们对内分泌 FGF 在人类新陈代谢中的作用的理解，并为糖尿病、肥胖、高胆固醇血症、结肠癌和慢性肾病等主要人类疾病的药物发现提供蓝图，其中大多数疾病对公众健康造成了巨大负担。 Aim III的结果将增强我们对FGFR和其他RTK致病突变作用机制以及整个RTK超家族酪氨酸激酶活性调节的理解。由于 RTK 的底物招募和磷酸化是 RTK 信号传导中的普遍事件，因此在 Aim IV 中获得的机制见解也将直接适用于整个 RTK 超家族。