Structural and Biochemical Basis of the Vitamin K cycle

描述(申请人提供)：哮喘是一种慢性肺部炎症性疾病，被认为完全由Th2细胞的产物介导。初步证据表明，过敏原诱导的呼吸道高反应性(AHR)的遗传易感性是由于上皮细胞对吸入性过敏原的不同反应所致。在易感动物中，上皮细胞衍生因子选择性地促进肺髓系树突状细胞(MDC)激活和产生Th17偏斜细胞因子。然而，目前尚不清楚上皮细胞如何增强MDC促进Th17反应的能力，或者Th17和Th2细胞如何合作加剧哮喘。其目的是：1)鉴定促进肺MDCs活化的上皮细胞因子；2)鉴定Th17应答在加重Th2相关病理过程中的作用；3)利用新的工具--来自协和杂交的重组近交系小鼠品系--鉴定新的哮喘易感基因。该提案中概述的研究和职业发展活动将使候选人能够实现其长期的职业目标，即成为一名独立的研究人员，专注于剖析遗传差异如何影响上皮细胞调节树突状细胞活性的能力，并促进Th17T细胞反应。为了获得必要的技能，候选人的K99研究将在玛莎·威尔斯-卡普的实验室进行，玛莎·威尔斯-卡普是哮喘遗传学和肺上皮培养技术领域的专家，在那里他将获得培养和操纵肺上皮细胞的专业知识 小鼠肺上皮细胞。此外，在指导阶段，候选人将通过在辛辛那提大学和杰克逊实验室接受高级培训，获得使用遗传和生物信息学方法识别新的哮喘易感基因的专业知识。这项培训对于完成为该奖项的独立阶段提出的研究至关重要。随后的研究将在小鼠和人类群体中剖析已识别的哮喘易感基因的功能，这将形成未来R01应用的基础，有助于向科学独立性的过渡。

DESCRIPTION (provided by applicant): Asthma is a chronic inflammatory disease of the lung thought to be mediated exclusively by the products of Th2 cells. Preliminary evidence suggests that genetic susceptibility to allergen-induced airway hyperresponsiveness (AHR) results from differential epithelial cell responsiveness to inhaled allergen. In susceptible animals, epithelial cell-derived factors selectively enhance the activation, and production of Th17-skewing cytokines by pulmonary myeloid dendritic cells (mDC). However, it is unclear how the epithelium enhances mDC capacity to promote Th17 responses, or how Th17 and Th2 cells collaborate to exacerbate asthma. The aims are: 1) to identify the epithelial cell factors responsible for promoting the activation of pulmonary mDCs, 2) identify the role of Th17 responses in exacerbating the Th2-associated pathology, and 3) identify novel asthma susceptibility genes using a novel tool, recombinant inbred mouse strains from the Collaborative Cross. The studies and career development activities outlined in this proposal will allow the candidate to achieve his long term career goal of becoming an independent investigator with a focus on dissecting how genetic differences influence the ability of epithelial cells to regulate dendritic cell activity, and promote Th17 T cell responses. To acquire the necessary skill set, the candidate's K99 research will take place in the lab of Marsha Wills-Karp, an expert in the field of asthma genetics, and pulmonary epithelial culture techniques, where he will acquire expertise in the culture and manipulation of murine pulmonary epithelium. Moreover, during the mentored phase, the candidate will acquire expertise in the use of genetic and bioinformatics approaches to identify novel asthma susceptibility genes, through advanced training at the University of Cincinnati, and Jackson Laboratories. This training will be essential for completing the research proposed for the independent phase of this award. Subsequent studies to dissect the function of identified asthma susceptibility genes in murine and human populations will form the basis of future R01 applications, assisting in the transition to scientific independence.