Regulation of Hedgehog Receptor Trafficking and Function

Hedgehog受体运输和功能的调控

• 批准号: 6460054
• 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/6460054
• 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.

描述（由申请人提供）：作为第一年的助理教授 儿科，申请人对发育遗传学和精炼感兴趣 作为学术领域的独立研究人员取得成功所需的技能 药这项研究提案的重点涉及探索刺猬 (Hh)途径，一个基本的信号级联负责早期胚胎 发育中的胎儿几乎每个器官系统的模式。Hh也 信号通路也在控制细胞生长和增殖中起作用， 成熟的生物体，而这一途径的突变会在易感的 组织中关于亚细胞定位的调节知之甚少， Hh受体复合物的运输或功能。特别感兴趣的是 Hh受体Patched（Ptc）和Hh配体本身似乎 不仅被其他蛋白质修饰，而且可能被其他蛋白质调节， 脂肪，特别是胆固醇。这五年的奖励将丰富 申请人的研究职业发展，通过调查 Hedgehog受体运输和功能发育的调节， 研究Hedgehog受体运输和功能的调节， 胆固醇和小窝蛋白-L。最初，共焦成像研究将是 用于评估受体的运输/亚细胞定位 复合物，包括受体内化Hh配体的能力。 将使用Caveline-1和胆固醇阻断剂的突变体构建体 确定Hh中每个组件所扮演的角色 受体运输和配体内化。荧光素酶报告 由Hh下游靶基因Gli组成的构建体将用于 测定Hh受体的功能。这将有助于确定Ptc的区域 和小窝蛋白是Hh受体功能的关键， 胆固醇在这个过程中的作用小窝蛋白-l缺失细胞系和小窝蛋白-l 将产生腺病毒载体，用于动态研究 caveolin-1在运输和功能中的作用。这项工作将与 导师实验室正在进行的关于caveolin-l基因敲除小鼠的研究， 检查发育缺陷和癌症形成。蒂莫西博士 汤普森的实验室，其主要重点是小窝蛋白-l在控制 细胞生长，分化和肿瘤发生，是一个很好的环境， 协助制定本提案。 由于无处不在的作用， Hh通路及其与癌症易感性的联系，该项目可能 提供临床应用，以了解异常的人类胎儿 细胞生长和分化的发育和控制。了解 这一途径的调节可能提示基因治疗的靶点， 发育中的胎儿或某些Hh相关癌症的治疗剂。