Genetic Regulation of IgE Glycosylation

项目摘要 过敏性疾病包括哮喘、湿疹和食物过敏在过去的几年里大幅增加。 几十年这些疾病影响了近5%的人口，每年的经济负担超过 八百亿美元。过敏反应可能危及生命。虽然我们已经了解了很多关于过敏性疾病 尽管过敏反应发生了，但关于过敏是如何发展的仍有很多未知之处。遗传易感性在 过敏性疾病的发生，但很少有明确的特定基因突变已知会导致过敏性疾病。 最近，我们在诱变小鼠中进行了过敏筛选，以发现调节 产生过敏原特异性IgE，介导过敏反应。在几种感兴趣的表型中， 苯那君的特点是IgE水平降低90%，对过敏反应有抵抗力，免疫力正常。 苯海拉明被归因于一个突变，一个产生用于糖基化的甘露糖的基因（将糖加到 蛋白质）、代谢和将蛋白酶靶向溶酶体。IgE糖基化对其结构很重要 和功能 因此（1），我们将通过鉴定细胞类型来确定苯那君突变如何影响IgE 通过体外细胞测定和条件性敲除小鼠模型的组合， 为了确定改变甘露糖的影响（2），我们将检查IgE糖基化的改变，稳定性， 功能沿着测量代谢和溶酶体运输的变化。评估…的重要性 在过敏性疾病中苯那君（3），我们将确定苯那君小鼠是否对IgE介导的 过敏反应和食物过敏。 成功完成后，这项研究的结果将揭示一个新的途径，重要的IgE 这将是一个有吸引力的治疗目标。我计划用这些成果来开展我的职业生涯， 这种小鼠品系，以了解如何改变IgE糖基化调节IgE的稳定性，清除和功能。 很少有其他研究者在探索这个话题，这将使我能够独立地研究这个问题到未来 与我现任导师的科学研究路线没有重叠 我的职业目标是成为一名独立的医生科学家，确定调节IgE的基因 生产，这可以在临床上转化为基因测试和潜在的治疗靶点。这个职业 发展奖将帮助我获得分子生物学，过敏症的动物模型，细胞免疫学， 和蛋白质组学与我的指导委员会一年两次的会议将指导早期的职业里程碑，包括 提交多份文件。机构赠款写作资源将用于成功竞争一个 R01获得独立。

Project Summary Allergic diseases including asthma, eczema, and food allergy have increased substantially over the last few decades. These diseases affect almost 5% of the population and having an annual economic burden of over 80 billion dollars. Allergic reactions can be life-threatening. While we have learned much about how an allergic reaction occurs, much remains unknown about how allergies develop. Genetic predisposition is important in developing allergies, but few defined mutations in specific genes are known to cause allergic disease. Recently, we conducted an allergy screen in mutagenized mice to discover genes that regulate production of allergen-specific IgE, which mediates allergic reactions. Among several phenotypes of interest, benadryl is marked by 90% reduced IgE levels and resistance to anaphylaxis with otherwise normal immunity. benadryl was ascribed to a mutation a gene that produces mannose used for glycosylation (adding sugars to proteins), metabolism and targeting proteases to the lysosome. IgE glycosylation is important for its structure and function. Therefore (1), we will determine how the benadryl mutation affects IgE by identifying the cell type responsible for low IgE through a combination of in vitro cell assays and conditional knock-out mouse models. To determine the effect of altered mannose (2), we will examine alterations to IgE glycosylation, stability, and function along with measuring changes to metabolism and lysosomal trafficking. To assess the importance of benadryl in allergic disease (3), we will determine whether benadryl mice are resistant to IgE mediated anaphylaxis and food allergy. Upon successful completion, the findings of this study will reveal a new pathway important for IgE production that would be an appealing therapeutic target. I plan to use these results to launch my career using this mouse strain to understand how altered IgE glycosylation regulates IgE stability, clearance and function. Few other investigators are exploring this topic, which will allow me to study this into the future independently without overlap with my current mentor’s line of scientific investigation. My career goal is to be an independent physician scientist determining the genes that regulate IgE production, which can be clinically translated to genetic tests and potential therapeutic targets. This career development award will help me gain skills in molecular biology, animal models of allergy, cellular immunology, and proteomics. Biannual meetings with my mentoring committee will guide early career milestones including submission of multiple papers. Institutional grant writing resources will be used to successfully compete for an R01 to gain independence.