Regulation of Hedgehog Receptor Trafficking and Function

Hedgehog受体运输和功能的调控

• 批准号: 6613777
• 负责人: HEIDI Eigenrauch KARPEN
• 金额: $ 12.59万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-19 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6613777
• 关键词: animal genetic material tag; biological signal transduction; caveolins; cell growth regulation; cell line; cholesterol; confocal scanning microscopy; developmental genetics; gene targeting; genetic regulation; genetically modified animals; human genetic material tag; immunoprecipitation; intermolecular interaction; laboratory mouse; neoplasm /cancer genetics; protein localization; protein structure function; protein transport; receptor; receptor binding; tissue /cell culture

DESCRIPTION (provided by applicant): As a first year Assistant Professor of Pediatrics, the applicant is interested in developmental genetics and refining the skills needed to achieve success as an independent researcher in academic medicine. The focus of this research proposal involves exploring the Hedgehog (Hh) pathway, a fundamental signaling cascade responsible for early embryonic patterning of nearly every organ system in the developing fetus. The Hh also pathway also plays a role in control of cell growth and proliferation in mature organisms, and mutations in this pathway cause cancer in susceptible tissues. Little is known about regulation of the subcellular localization, trafficking or function of the Hh receptor complex. Of particular interest is the fact that the Hh receptor Patched (Ptc), and the Hh ligand itself, seem to be modified, and potentially regulated not only by other proteins, but also by lipids, particularly cholesterol. This five-year award will provide enrichment of the applicant's research career development through investigating the regulation of Hedgehog receptor trafficking and function development through investigating the regulation of Hedgehog receptor trafficking and function by cholesterol and caveolin-l. Initially, confocal imaging studies will be employed to assess trafficking/subcellular localization of the receptor complex, including the ability of the receptor to internalize the Hh ligand. Mutant constructs of caveline-1 and cholesterol blocking agents will be used to identify the individual roles played by each of these components in Hh receptor trafficking and ligand internalization. A luciferase reporter construct, comprised of a Hh downstream target gene, Gli, will be used to assay function of the Hh receptor. This will help determine the regions of Ptc and caveolin that are critical for Hh receptor function and better define the role of cholesterol in this process. Caveolin-l null cell lines and caveolin-l adenoviral vectors will be generated for dynamic studies of the role of caveolin-l in trafficking and function. This work will be integrated with ongoing work in the mentor's lab on a caveolin-l knockout mouse, which will be examined for developmental defects and cancer formation. Dr. Timothy Thompson's lab, whose primary focus is the role of caveolin-l in control of cell growth, differentiation and tumorigenesis, is an excellent environment to aid in the development of this proposal. Because of the ubiquitous role of the Hh pathway, and its link to cancer predisposition, this project may provide clinical application to the understanding of aberrant human fetal development and control of cell growth and differentiation. Understanding the regulation of this pathway may suggest targets for gene therapy in the developing fetus or therapeutic agents for certain Hh-related cancers.

简介(由申请人提供)：担任大学第一年助理教授 小儿科，申请人对发育遗传学和精炼感兴趣 作为一名学术领域的独立研究员取得成功所需的技能 医药。这项研究计划的重点是探索刺猬 (HH)途径，早期胚胎的基本信号级联反应 发育中的胎儿几乎每个器官系统都有图案。HH还 途径也在控制细胞生长和增殖方面发挥作用。 成熟的生物体，而这一途径的突变会导致易感人群的癌症 纸巾。对亚细胞定位的调节知之甚少， HH受体复合体的运输或功能。特别值得关注的是 事实是，HH受体补丁(PTC)和HH配体本身似乎 不仅受到其他蛋白质的修饰和潜在的调节，而且还受到 脂类，尤其是胆固醇。这项为期五年的奖励将提供丰富的 通过调查申请人的研究生涯发展情况 Hedgehog受体转运和功能发育的调控 Hedgehog受体转运和功能调控的研究 胆固醇和小窝蛋白-L。最初，共焦成像研究将是 用来评估受体的运输/亚细胞定位 复合体，包括受体内化HH配体的能力。 卡维林-1和胆固醇阻滞剂的突变结构将被使用 确定这些组件中的每个组件在HH中扮演的单独角色 受体运输和配基内化。一名荧光素酶记者 由HH下游靶基因Gli组成的构建将用于 HH受体的检测功能。这将有助于确定PTC的区域 和小窝蛋白，对HH受体功能至关重要，并更好地定义 胆固醇在这一过程中的作用。洞穴-L空细胞系和洞穴-L 腺病毒载体的产生将用于动态研究 卡维林-L在贩卖中的作用和作用。这项工作将与 导师实验室正在进行一种洞穴-L基因敲除小鼠的工作，这将是 检查发育缺陷和癌症形成。蒂莫西医生 汤普森的实验室，其主要焦点是洞穴-L在控制 细胞的生长、分化和肿瘤的发生是一个极好的环境 帮助制定这项提案。因为无处不在的角色 HH途径及其与癌症易感性的联系，该项目可能 为了解人类畸形胎儿提供临床应用 细胞生长和分化的发育和控制。了解 对这一途径的调节可能提示基因治疗的靶点 开发胎儿或治疗某些HH相关癌症的药物。