Establishment and Application of a Library of Site-Specific Glycosylation Pattern

描述(由申请人提供)：我已经制定了一个职业发展战略，在关键领域的课程工作和密集的科学发展之间取得平衡。我的指导团队在事业、阶段和纪律方面都很平衡。我将通过他们的指导进一步发展我的技术技能和科学知识。我的两位导师(李博士和摩西博士)从事高度翻译的研究。此外，报名参加基金撰写和领导力培训课程将增强我实现并成功实现成为一名非常成功的独立研究人员的职业目标的能力。如果我的事业成功地过渡到独立，我希望利用我在糖蛋白组学、小分子定量和定量蛋白质组学方面的培训，开发新的策略来确定特定部位的糖基化模式分析。希望将这些策略应用于阐明该病的病理和病因。 涉及糖基化的复杂疾病。通过接触导师的科学和翻译专业知识，我将接受的培训对我作为一名独立研究人员的成功至关重要。这项建议的总体科学目标是解决蛋白质糖基化分析中的关键挑战，并在尿路感染的情况下更彻底地确定蛋白质糖基化，尿路感染是一种已知受糖基化影响的疾病。糖基化是一种非常重要的蛋白质修饰，但研究还不够深入。这项建议通过执行特定部位的糖基化图谱(SSGP)分析来研究尿蛋白糖基化的新细节。不幸的是，阻碍SSGP成为常规的重大挑战仍然存在，而且SSGP目前的吞吐量很低。我们建议通过开发SSGP文库来改进SSGP分析，使未来的SSGP分析更容易、更快和更灵敏。我们将进一步利用该文库建立健康个体的健康基线尿SSGP。此外，糖基化已知会影响尿路感染(UTI)。因此，将开展一项有重点的研究，在反复尿路感染的个体中识别SSGP，并寻找新的尿液保护因素。本研究的目的是：目的1：测定复发性尿路感染患者和健康对照组的尿糖蛋白SSGP；目的2：建立健康人尿中SSGP的综合文库。我在这些科学目标上得到了极好的研究环境的支持。波士顿儿童医院(BCH)一直是美国最好的儿童医院之一。在医院附近有4栋高层建筑，居住着来自BCH的研究人员。BCH有数十位国际公认的科学家，其中包括该项目的主要导师玛莎·摩西博士。两位科学家因在波士顿大学做出的贡献而获得诺贝尔奖。重要的是，在泌尿科内有一个基本的、分类良好的生物库计划，这将是这项研究成功的关键。

DESCRIPTION (provided by applicant): I have developed a career development strategy which balances course work in critical areas with intensive scientific development. My mentoring team is well-balanced by career stage and discipline. I will further develop my technical skills and scientific knowledge through their mentorship. Two of my mentors (Dr. Lee and Dr. Moses) engage in highly translational research. Additionally, enrollment in courses on grant writing and leadership training will enhance my ability to achieve and succeed in my career goal of becoming a highly successful independent researcher. If I am successful in my career transition to independence, I hope to use my training in glycoproteomics, small molecule quantitation, and quantitative proteomics to develop novel strategies to determine site-specific glycosylation pattern analysis. I hope to apply these strategies to the elucidation of the pathology and etiology of complex diseases involving glycosylation. The training I will receive through exposure to my mentors' scientific and translational expertise will be critical for my success as an independent researcher. The overall scientific goal of this proposal is to address critical challenges in the analysis of protein glycosylation, and determine protein glycosylation more thoroughly in the context of urinary tract infection, a condition known to be affected by glycosylation. Glycosylation is a critically important protein modification, and is understudied. This proposal investigates urinary protein glycosylation to new detail by performing a site- specific glycosylation profile (SSGP) analysis. Unfortunately, significant challenges remain preventing SSGP from becoming routine, and SSGP is currently low-throughput. We propose to improve SSGP analysis by developing a library of SSGP to make future SSGP analyses more accessible, faster and more sensitive. We will further use this library to establish the healthy baseline urinary SSGP among healthy individuals. In addition, glycosylation is known to affect urinary tract infections (UTI). Therefore a focused study will be performed to identify SSGP among individuals who have recurrent urinary tract infections, and to seek out new protective factors in urine. The aims of the study are: Aim 1: Determine urinary glycoprotein SSGP from patients with recurrent UTI and healthy controls; and Aim 2: Create a comprehensive library of SSGP in the urine of healthy individuals. I am extremely well supported in these scientific goals by an excellent research environment. Boston Children's Hospital (BCH) is consistently ranked among the finest children's hospitals in the US. There are 4 high-rise buildings housing researchers from BCH in the immediate vicinity of the hospital. There are dozens of internationally recognized scientists at BCH, including the primary mentor on this project, Dr. Marsha Moses. Two scientists have been recognized with the Nobel prize for contributions made at BCH. Importantly, there is an essential, well-categorized biobanking program within the department of Urology which will be essential to the success of this study.