The Modulation of Eotaxin Expression by Statins: Implications for Asthma Therapy

描述(由申请人提供)：这份K08提案的主要目标是帮助我成为一名拥有NIH R01级资金的独立调查员。我选择了吴琳教授作为主要导师，他是该领域的全国领导者，在以前的导师中有着成功和长期的记录。作为一名研究员，我最初在尼古拉斯·肯扬博士的实验室工作，并学会了使用卵清蛋白小鼠模型。这导致了对小鼠和人类呼吸道上皮细胞的初步实验，以确定他汀类药物是否改善过敏性炎症和上皮损伤，如果是，是通过什么机制。2009年，我被授予美国胸科学会(ATS)职业发展奖，以研究辛伐他汀通过抑制甲氧戊酸(MA)途径对气道重塑的影响。从那时起，我还在当地以及与其他机构的同事建立了几次合作，从而获得了一笔资助赠款(TRDRP)和几份出版物。在我担任研究员和博士后期间，我参加了区域会议和ATS年度国际会议上的研究生课程和研讨会。作为目前的K12(KL2)学者，我通过我们的代谢组学核心帮助共同开发了测量和量化肺组织中他汀类药物和MA代谢物的新方法。2012年，我被任命为助理教授常驻教授，这是一个专注于研究的学术系列。这一任命是值得的，并表明了我们学院对我的职业发展的承诺。这份K08提案建立在我之前工作的基础上，并将为确保未来的R01资金奠定基础。环境：加州大学戴维斯分校的整体研究环境非常出色，非常适合我的研究和职业发展目标。比较呼吸生物学和医学中心为研究人员、研究生和临床研究员之间提供一流的、持续的和丰富的智力交流。生物科学学院每周举办分子生物学研讨会，有来自分子生物学所有领域的世界级演讲者参加。加州大学戴维斯分校通过医学院和兽医学院以及加州国家灵长类动物研究中心，对肺部研究的博士后和博士后研究实习生进行了长期的培训。这些学校的肺病研究尤其强大，许多由NIH资助的研究人员和公认的专家在附近工作。我们的临床和转化科学中心(CTSC)和CTSC临床研究中心提供持续的研究基础设施支持，这是我在加州大学戴维斯分校非常熟悉的资源。职业目标：我的首要职业目标是在学术医学中心成为一名独立且富有成效的内科科学家，在哮喘等呼吸道疾病方面拥有临床和基础科学专业知识。我的愿景是领导一个实验室，研究与呼吸道疾病和其他肺部疾病相关的呼吸道上皮生物学和致病机制，并开发创新的治疗方法。最终，我还将指导和培训住院医生、研究员、研究生和初级教员在他们的研究和学术生涯中。职业发展计划：该计划有4个培训领域(为期3年的授权期)：研究技能、课程和培训、手稿和补助金。我已经概述了一个个性化的教育计划，以不同的百分比努力突出我的研究计划的发展。我将把至少75%的时间用于研究，这与25%的临床时间很好地吻合，包括重症哮喘诊所(1/2天/周)，住院轮换(6周/年)，以及行政职责/每周研讨会/讲座。这项计划包括一个指导团队和咨询委员会，并定期计划召开会议来评估我的进展。研究项目：尽管目前有治疗方法，但一些哮喘患者的哮喘症状仍未得到有效控制。嗜酸性粒细胞趋化因子(-1，2，3)是一种有效的嗜酸性粒细胞特异性趋化因子，在严重哮喘中起重要作用。尤其是嗜酸性粒细胞趋化蛋白-3与显著的呼吸道和全身性过敏性炎症以及哮喘易感性的增加密切相关。观察性研究和小型临床试验表明，降脂他汀类药物可能会改善肺部健康。在我们的小鼠模型中，我们发现辛伐他汀(Sim)可以减轻嗜酸性呼吸道炎症、IL13/IL4的产生和呼吸道的高反应性。使用原代正常人支气管上皮细胞，Sim抑制基础和IL13诱导的eoaxin-2和-3的表达和STAT6的磷酸化，而不改变eoaxin-3mRNA的稳定性。由于嗜酸性粒细胞趋化蛋白的表达受IL13诱导的JAK/STAT6的直接转录调控，我假设辛伐他汀(1)抑制呼吸道上皮细胞 通过调节IL13诱导的JAK/STAT6信号，在转录水平上表达eoaxin-2和-3基因，以及(2)减少严重过敏性哮喘患者的病情恶化。目的：1)检测Sim是否在转录水平上同时抑制基础和IL13诱导的嗜酸性粒细胞趋化因子基因表达。目的2)检测Sim是否抑制IL13诱导的JAK/STAT6信号通路。目的3)确定SIM(A)是否能减少TH2过敏性炎症介质在支气管上皮细胞中的表达，以及(B)是否能减轻急性加重和改善肺功能。我将进行一项为期30周的前瞻性、双盲、交叉的早期II期临床试验，将重度哮喘患者随机分为安慰剂和SIM两组，间隔12周，并给予标准护理吸入皮质类固醇和长效？-激动剂。

DESCRIPTION (provided by applicant): The primary goal of this K08 proposal is to help me become an independent investigator with NIH R01 level funding. I have chosen as primary mentor Professor Reen Wu, who is a national leader in his field with a successful and long track record of prior mentees. As a fellow I initially worked in the laboratory of Dr. Nicholas Kenyon and learned to use the ovalbumin mouse model. This led to preliminary experiments in mouse and human airway epithelial cells to determine whether statins ameliorate allergic inflammation and epithelial injury, and if so, by what mechanisms. In 2009, I was awarded an American Thoracic Society (ATS) Career Development Award to study simvastatin's effects on airway remodeling via mevalonate (MA) pathway inhibition. Since then I also formed several collaborations both locally and with colleagues at other institutions that have led to a funded grant (TRDRP) and several publications. Throughout my fellowship and post-doctoral years I attended post-graduate courses and seminars at regional meetings and at the ATS annual international conferences. As a current K12 (KL2) scholar, I helped co-develop novel methodologies via our Metabolomics Core for measuring and quantifying the statins and MA metabolites in lung tissue. In 2012, I was appointed as an Assistant Professor in Residence, which is a research-focused academic series. This appointment is rewarding and demonstrates our Institution's commitment to my career development. This K08 proposal builds on my prior work and will lay the foundations to secure future R01 funding. Environment: The overall research environment at U.C. Davis is outstanding and well-suited for my research and career development goals. The Center for Comparative Respiratory Biology & Medicine provides for first class continuous and rich intellectual exchange among research faculty, graduate students, and clinical fellows. The College of Biological Sciences holds weekly Seminars in Molecular Biology that feature world- class speakers from all fields of molecular biology. U.C. Davis has a long history of training pre- and post- doctoral research trainees in pulmonary research through the Schools of Medicine and Veterinary Medicine, and California National Primate Research Center. Lung research in these schools is particularly strong with many NIH-funded investigators and recognized experts working in close proximity. Our Clinical and Translational Science Center (CTSC) and CTSC Clinical Research Center provide for continuous research infrastructure support, a resource I am very familiar with here at U.C. Davis. Career Goals: My overarching career goal is to become an independent and productive physician-scientist at an academic medical center, with clinical and basic science expertise in airway diseases such as asthma. My vision is to lead a laboratory that will investigate airway epithelial biology and pathogenic mechanisms relevant to airway diseases and perhaps other lung diseases, and develop innovative and novel therapies. Eventually, I will also mentor and train residents, fellows, graduate students, and junior faculty in their research and academic careers. Career Development Plan: This plan has 4 domains of training (during a 3-year award period): Research Skills, Coursework and Training, Manuscripts, and Grants. I have outlined an individualized educational plan with different percent efforts to highlight the development of my research program. I will devote at least 75% of my time to research, which fits well with the 25% clinical time including severe asthma clinic (1/2 day/week), in- patient rotations (6 weeks/year), and administrative duties/weekly seminars/lectures. This plan includes a mentoring team and Advisory Committee with regular planned meetings to gauge my progress. Research Project: Asthma symptoms remain poorly controlled in some asthmatics despite current treatments. The eotaxins(-1,2,3) are potent TH2 eosinophil-specific chemokines, important in severe asthma. Eotaxin-3 in particular is strongly associated with marked airway and systemic allergic inflammation, and increased asthma susceptibility. Observational studies and small clinical trials suggest that the lipid-lowering statin drugs may improve lung health. In our mouse model, we found that simvastatin (Sim) attenuates eosinophilic airway inflammation, IL13/IL4 production, and airway hyperreactivity. Using primary normal human bronchial epithelial cells, Sim suppressed basal and IL13-induced eotaxin-2 and -3 expression and STAT6 phosphorylation, without altering eotaxin-3 mRNA stability. Since eotaxin expression is under direct IL13-induced JAK/STAT6 transcriptional regulation, I hypothesize that simvastatin (1) inhibits airway epithelial eotaxin-2 and -3 gene expression at the transcriptional level through modulation of IL13-induced JAK/STAT6 signaling, and (2) reduces exacerbations in patients with severe allergic asthma. Specific Aims: Aim 1) To test whether Sim inhibits both basal and IL13-induced eotaxin gene expression at the transcriptional level. Aim 2) To test whether Sim inhibits the IL13-induced JAK/STAT6 signaling pathway. Aim 3) To determine whether Sim (A) decreases mediators of TH2 allergic inflammation in bronchial epithelial cells, and (B) reduces acute exacerbations and improves lung function. I will perform a 30-week prospective, double-blinded, cross-over early Phase II clinical trial and randomize severe asthma patients to both placebo and Sim for 12 week intervals, in addition to standard-of-care inhaled corticosteroid and long-acting ?-agonist.