Functional Genomics Approach to the Role of the JAK/STAT and MAPK Pathway in CBD

描述(由申请人提供) 这项提案概述了一项全面的三年培训计划，候选人将成为学术、肺部和重症监护医学领域的独立科学家。应聘者在华中科技大学同济医学院获得博士学位，在德国乌尔姆大学获得医学博士学位。她的研究重点是为急性髓系白血病(AML)患者开发新的免疫治疗策略。随后，她在科罗拉多大学丹佛分校皮肤科接受博士后培训。她的研究重点是激活的记忆T淋巴细胞和抗原提呈细胞之间的细胞-细胞免疫突触相互作用。此外，她获得了他在国家犹太人健康(NJH)的第一个学术职位，在环境和职业健康科学(DEOHS)的医学系，研究由于暴露于铍而导致的肉芽肿性肺部疾病慢性铍疾病(CBD)。在她被任命后不久，她成功地通过UCD肺科获得了K12机构颁发的职业发展奖。这笔助学金为她提供了加州大学学院讲师级别的教职。在那里，她精通基因组学和CBD的免疫致病机制。 由环境科学、遗传学、基因组学和生物信息学专家、卫生部主任迈尔博士担任主要导师，该计划在三个基本领域为候选人提供额外的培训：1)遗传学、基因组学、生物统计学和更新方面的教育 基本的实验室技能；2)在监督下准备研究出版物；3)指导她独立研究提案的开发。其他指导由共同导师戴博士提供，戴博士是一位具有丰富先前指导经验的肺部先天免疫和炎症专家，以及一个科学家专家小组。 该项目将在UCD和NJH进行，这两个地方都为培养科学家提供了出色的环境。它提供最先进的研究和临床设施，提供绝佳的教育和科学思想交流机会，并坚定地致力于候选人作为独立研究人员的发展。 这项科学提案探索了一种新的假设，即CBD患者对免疫相关基因的不同调节最终会导致免疫功能异常，并增加BE的易感性。具体地说，中心假设是Be暴露导致JAK/STAT通路和MAPK通路的不适当激活，并参与CBD的发病。因此，本研究的目的是通过深入分析CBD中差异表达的候选基因，深入了解CBD的发病机制和致病途径。其具体目的是：1)与铍暴露的BES和Be暴露对照相比，证明JAK/STAT通路和MAPK通路相关基因在CBD中被不适当地激活，并利用qRT-PCR验证这些在微阵列分析中发现的潜在的重要CBD候选基因。2)通过FLP-in体外表达分析系统检测JAK/STAT和MAPK通路候选基因产物的过度表达或siRNA“敲除”分析基因产物的减少，从功能上评价JAK/STAT通路和MAPK通路候选基因在CBD发病机制中的作用。3)研究差异基因是否与CBD(n=100)和BES(n=100)在更大的独立人群中存在关联，从而揭示CBD潜在的新的生物标志物。从这些数据中获得的见解将大大增加对CDB分子机制的了解，并可能导致开发潜在的新生物标记物和新的研究靶点，作为潜在的CBD治疗方法。 公共卫生相关性：令人兴奋的初步研究表明，与暴露在未患病的对照组相比，CBD和BES的外周血细胞显示出与免疫功能和这些过程的发病机制相关的不同基因表达谱。这一建议是通过深入分析CBD中差异表达的候选基因，重点关注Th1免疫反应的途径，来深入了解CBD的重要机制和致病途径。这一结果有助于我们更好地了解CBD的发病因素、影响预后的因素以及治疗的靶点，可作为暴露相关免疫性疾病的模型，并有可能应用于其他环境诱发的疾病。

DESCRIPTION (provided by applicant) This proposal outlines a comprehensive three year training plan for the candidate to become an independent scientist in academic pulmonary and critical care medicine. The candidate earned her Ph.D. from the Tongji Medical College, Huazhong University of Science &Technology (HUST) and M.D. from University of Ulm, Germany. Her research focused on the development of novel immunotherapeutic strategies for patients with acute myeloid leukemia (AML). She subsequently received postdoctoral training in the Department of Dermatology, University of Colorado Denver (UCD). Her research was highly focused on the cell-cell immunological synapse interactions between activated memory T lymphocytes and antigen presenting cells. In addition, she received his first academic appointment at National Jewish Health (NJH) in the Division of Environmental & Occupational Health Sciences (DEOHS) in the Department of Medicine studying the granulomatous lung disease chronic beryllium disease (CBD), which results from exposure to beryllium. Soon after her appointment, she was successful in obtaining a career development award from an institutional K12 through the Pulmonary Division, UCD. This grant provided her with a faculty appointment at an Instructor level at UCD. There, she developed proficiency in genomics, and the immunopathogenic mechanisms of CBD. With Dr. Maier, an expert in environmental sciences, genetics, genomics and bioinformatics, head of DEOHS, serving as the primary mentor, this program provides the candidate additional training in three essential areas: 1) education in genetics, genomics, biostatistics and updates in essential laboratory skills; 2) supervised preparation of research publications; 3) mentored development of her independent research proposal. Additional mentoring is provided by co-mentor Dr. Day, Ph.D., an expert in pulmonary innate immunity and inflammation with extensive prior mentoring experience, and an expert panel of scientists. This program will take place at UCD and NJH, both of which provide outstanding environment for training scientists. It offers state-of-the-art research and clinical facilities, outstanding opportunities for education and the exchange of scientific ideas, and strong commitment to the candidate's development as an independent investigator. The scientific proposal explores the novel hypothesis that patients with CBD have differential regulation of immune-related genes that ultimately leads to abnormalities in immune function and heightened susceptibility to be. Specifically, the central hypothesis is that Be-exposure results in the inappropriate activation of the JAK/STAT pathway and MAPK pathway and contributes to CBD pathogenesis. Therefore, the goal of this study is to gain insight into the mechanisms and pathogenic pathways important in CBD by conducting an in- depth analysis of the differentially expressed candidate genes in CBD. Its specific aims are: 1) To demonstrate that the JAK/STAT pathway and MAPK pathway related genes are inappropriately activated in CBD compared to BeS and Be-exposed controls with beryllium-exposure, validating these potentially important CBD candidate gene identified in the microarray analysis using qRT-PCR. 2) To functionally assess the role of the candidate genes in the JAK/STAT pathway and MAPK pathway in CBD pathogenesis by assessing either the over- expression of the gene product using the Flp-in expression analysis system in vitro, or the reduction of the gene product using siRNA "knock down" analysis. 3) To investigate whether the differentially genes are associated with CBD (n=100) and BeS (n=100) in a larger separate population and thus reveal potential novel biomarkers for CBD. Insights garnered from these data will significantly add to the understanding of molecular mechanisms of CDB and may lead to development of potential novel biomarker and new targets for study as potential therapeutic approaches for CBD. Public Health Relevance: Exciting preliminary studies show that the peripheral blood cells from CBD and BeS demonstrate differential gene expression profiles relevant to the immune function and pathogenesis of these processes, compared to Be-exposed non-diseased controls. This proposal is to gain insight into the mechanisms and pathogenic pathways important in CBD by conducting an in-depth analysis of the differentially expressed candidate genes in CBD focusing on a pathway that is to a Th1 immune response. The results may improve our understanding of factors involved in the development of CBD, aspects which determine prognosis, and targets for therapy, serves as a model of exposure-related immunologic disease with potential application to other environmentally induced diseases.