Molecular Mechanisms of iNOS Degradation

描述（由申请人提供）：该候选人是最近独立的内科科学家，并表现出对生物医学研究的强烈承诺。他的长期职业目标是在气道炎症的分子机制方面开展强有力的研究项目。他的近期研究目标是了解诱导型一氧化氮合酶（iNOS）在气道细胞中调节NO合成的分子机制，并设计控制方法。一个成功的R01申请使候选人准备继续他目前的研究，启动新的项目，获得新的技能和建立新的合作。需要K02奖来减少候选人目前任命的临床成分，从而允许一段时间的密集研究重点，以促进他的科学发展，扩大他的潜力，做出重大的研究贡献。候选人在一个高度支持的环境中，有优秀的导师和大量的智力发展机会。他是新成立的“炎症生物学”中心的教员。候选人的实验室位于中心及其核心实验室内，该核心实验室由新装修的最先进的开放实验室设计设施组成，占地20,000平方英尺。该中心的研究环境为研究人员提供了一个动态的临界质量，为候选人的研究提供了一个协同的环境。核心研究项目（为期5年的R01资助项目）的总体目标是了解气道上皮细胞中iNOS降解的机制。它包括四个具体目标：1)鉴定人类iNOS降解途径。2)表征泛素化在iNOS降解中的可能作用。3)阐明人iNOS的亚细胞定位。4)变构抑制剂对iNOS降解的调节分析。该候选人正在通过合作研究解决三个额外的具体目标：1)确定iNOS泛素化的特定位点。II)阐明磷酸化在iNOS泛素化中的作用。III)鉴定特异性泛素连接酶（E3）负责iNOS泛素化。研究将在上皮细胞系和原代气道支气管上皮细胞中进行。提出这些研究的理由是，一旦了解了iNOS的降解机制，就可以设计治疗策略来改变这些途径并调节iNOS的降解。这些研究结果将增加我们对iNOS调控的细胞过程的理解，从而为未来针对哮喘等气道炎症疾病中NO合成控制的研究奠定基础。

DESCRIPTION (provided by applicant): The candidate is a recently independent physician-scientist who has shown strong commitment to biomedical research. His long-term career goal is to develop a strong research program in molecular mechanisms of airway inflammation. His immediate research goals are to understand the molecular mechanisms of regulation NO synthesis by inducible nitric oxide synthase (iNOS) in airway cells and devise methods to control it. A successful R01 application has the candidate poised to continue his current studies, initiate new programs, acquire new skills and build new collaborations. The K02 award is needed to reduce the clinical component of the candidate's current appointment, thus allowing a period of intensive research focus to foster his scientific development and expand his potential to make significant research contributions. The candidate is in a highly supportive environment with excellent mentors and numerous opportunities for intellectual development. He is a faculty member of a newly established center for "Biology of Inflammation". The candidate's laboratory is within the center and its core laboratories, which comprise 20,000 sq, ft. of newly renovated state-of-the-art-open lab design facilities. The center's research environment offers a dynamic critical mass of investigators providing a synergistic environment for the candidate's research. The overall objective of the core research project (a five-year R01 funded 8/1/02) is to understand the mechanisms of iNOS degradation in airway epithelial cells. It consists of four specific aims: 1) Identification of human iNOS degradation pathway. 2) Characterization of possible role of ubiquitination in iNOS degradation. 3) Elucidation of the subcellular localization of human iNOS. 4) Analysis of modulation of iNOS degradation by allosteric inhibitors. The candidate is addressing three additional specific aims through collaborative studies: I) Determination of specific sites of iNOS ubiquitination. II) Elucidation of Role of phosphorylation in iNOS ubiquitination. III) Identification of specific ubiquitin ligase (E3) responsible for iNOS ubiquitination. Studies will be conducted in epithelial cell lines and in primary airway bronchial epithelial cells. The rational for the proposed studies is that once the degradation mechanisms of iNOS are understood, therapeutic strategies can be designed to alter these pathways and modulate iNOS degradation. The results of these studies will increase our understanding of the cellular process of iNOS regulation and thus lay the groundwork for future studies aiming at controlling NO synthesis in diseases of airway inflammation such as asthma.