HEPATOCYTE GROWTH FACTORS

肝细胞生长因子

• 批准号: 2139537
• 负责人: WALLACE LEE MCKEEHAN
• 金额: $ 19.54万
• 依托单位: TEXAS A&M AGRILIFE RESEARCH
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-02-01 至 1998-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2139537
• 关键词: HTC cell; biological signal transduction; cell growth regulation; cell type; chemical conjugate; fibroblasts; growth factor receptors; heparan sulfate; immunochemistry; laboratory rat; ligands; liver cells; liver regeneration; protein isoforms; protein purification; protein tyrosine kinase; receptor expression; recombinant DNA; site directed mutagenesis; transforming growth factors

Understanding the molecular mechanisms underlying the order and precision of compensatory liver regeneration is essential for understanding and intervention in liver carcinogenesis and toxicology as well as development of strategies for liver cell transplantation and gene therapy. Increasing evidence indicates that liver regeneration is orchestrated by activation and repression of the activity of multiple cytokines within the liver rather than external hormones. Of the polypeptide regulators endogenous to the local liver environment, the FGF family of eight ligands and their receptors (FGF-R) (four genes, 16 splice variants resulting in >100 isoforms) has large potential for transient regulation of growth and function in both parenchymal and non-parenchymal cells. This continuation project will isolate and characterize a potentially novel parenchymal cell-specific member of the FGF ligand family extracted directly from human hepatoma cells and perfused liver in vivo. Expression and molecular basis for FGF-1-specificity and specificity in signal transduction of FGF- R4 isoforms, a major FGF-R gene expressed in liver, will be determined. Expression of splice variants of the parenchymal cell-specific FGF-R2 gene, interaction with the FGF-R4 isoforms and their-role in signal transduction will be determined in resting and regenerating liver and derived cells. Ligand-, tyrosine kinase receptor- and cell type-specific natural heparan sulfate proteoglycan components of the FGF-R complex will be isolated and characterized. A potentially novel nuclear role of a specific complex of FGF-1 and a fragment of its receptor will be investigated. The temporal expression of the TGF-beta receptor and two novel serine/threonine kinase receptors during liver regeneration in parenchymal and nonparenchymal cells will be determined. FGF-toxin conjugates will be employed to probe the role of FGF ligands and receptor isotypes in regenerating liver in vivo. The project employs combined rigorous experimental approaches which include protein micropurification, immunochemistry, recombinant technologies (bacterial, baculoviral-insect cells, transfected mammalian cells) and site-directed mutagenesis to achieve the goals of the project.

了解顺序和精度的分子机制 补偿性肝脏再生对于理解和 干预肝癌发生和毒理学以及发育 肝细胞移植和基因疗法的策略。 增加 证据表明肝脏再生是通过激活而策划的 和抑制肝内多种细胞因子的活性 而不是外部激素。 多肽调节剂内源性 到当地的肝脏环境，FGF家族八个配体及其 受体（FGF-R）（四个基因，16个剪接变体，导致> 100 同工型）具有巨大的生长和 在实质和非核细胞中的功能。 这个延续 项目将隔离并表征潜在的新型实质 直接从FGF配体家族中提取的细胞特异性成员 人肝癌细胞和体内灌注肝。表达和分子 FGF-1特异性和FGF信号转导特异性的基础 R4同工型将确定在肝脏中表达的主要FGF-R基因。 实质特异性FGF-R2的剪接变体的表达 基因，与FGF-R4同工型的相互作用及其在信号中的角色 转导将在静止和再生肝脏中确定，并确定 衍生细胞。 配体 - 酪氨酸激酶受体和细胞类型特异性 FGF-R复合物的天然硫酸乙酰肝素蛋白聚糖成分将 被隔离和表征。 一个潜在的新型核作用 FGF-1及其受体片段的特定复合物将是 调查。 TGF-β受体的时间表达和两个 肝脏再生期间新型丝氨酸/苏氨酸激酶受体 将确定实质性和非实质细胞。 FGF毒素 偶联物将被用来探测FGF配体和受体的作用 在体内再生肝脏中的同种型。 该项目联合使用 严格的实验方法，包括蛋白质微纯化， 免疫化学，重组技术（细菌，杆状病毒启示 细胞，转染的哺乳动物细胞）和位于位置的诱变为 实现项目的目标。