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。 几十年了。这些疾病影响了近5%的人口，每年造成超过5%的经济负担 800亿美元。过敏反应可能危及生命。虽然我们已经学到了很多关于过敏的知识 反应发生时，过敏是如何发展的仍有许多未知之处。遗传易感性在 发生过敏，但很少有特定基因的明确突变会导致过敏性疾病。 最近，我们对突变小鼠进行了过敏筛查，以发现调节 产生过敏原特异性的IgE，它介导过敏反应。在几种感兴趣的表型中， 苯那曲利的特点是IgE水平降低90%，对过敏反应的抵抗力，其他方面免疫正常。 苯那君被归因于一种突变，一种产生用于糖基化的甘露糖的基因(向 蛋白质)、新陈代谢和针对溶酶体的蛋白水解酶。IgE糖基化对其结构有重要作用 和功能。 因此(1)，我们将通过鉴定细胞类型来确定苯乙酰突变对IgE的影响 通过体外细胞分析和条件性基因敲除小鼠模型的组合，导致低IgE。 为了确定甘露糖改变的效果(2)，我们将检查IgE糖基化、稳定性和 功能，同时测量新陈代谢和溶酶体运输的变化。评估…的重要性 苯那曲在变态反应性疾病中的作用(3)，我们将确定苯那君小鼠是否对IgE介导的抗体产生抵抗力 过敏反应和食物过敏。 成功完成后，这项研究的发现将揭示一条对IgE重要的新途径 这将是一个吸引人的治疗目标。我计划利用这些结果开始我的职业生涯，使用 这种小鼠品系了解改变的IgE糖基化如何调节IgE的稳定性、清除性和功能。 很少有其他研究人员在探索这个话题，这将使我能够在未来独立研究这个问题 不会与我目前导师的科学研究路线有重叠。 我的职业目标是成为一名独立的内科科学家，研究调节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。这份职业 发展奖将帮助我获得分子生物学、过敏动物模型、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我早期的职业里程碑，包括 提交多篇论文。机构赠款撰写资源将用于成功竞争 R01以获得独立。

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.