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，平衡在关键领域的课程工作与密集的科学发展。我的指导团队在职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）从事高度转化的研究。此外，在补助金写作和领导力培训课程的招生将提高我的能力，实现和成功，在我的职业目标，成为一个非常成功的独立研究员。如果我在我的职业生涯成功过渡到独立，我希望利用我在糖蛋白质组学，小分子定量和定量蛋白质组学的培训，开发新的策略，以确定位点特异性糖基化模式分析。我希望将这些策略应用于病理和病因的阐明 涉及糖基化的复杂疾病。我将通过接触我的导师的科学和翻译专业知识接受的培训将是我作为一个独立研究人员成功的关键。 该提案的总体科学目标是解决蛋白质糖基化分析中的关键挑战，并在尿路感染的背景下更彻底地确定蛋白质糖基化，这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，并且研究不足。该提案通过进行位点特异性糖基化谱（SSGP）分析来研究尿蛋白糖基化的新细节。不幸的是，仍然存在重大挑战，阻止SSGP成为常规，SSGP目前是低吞吐量的。我们建议通过开发SSGP库来改进SSGP分析，以使未来的SSGP分析更容易获得，更快，更灵敏。我们将进一步使用该文库建立健康个体的健康基线尿SSGP。此外，已知糖基化会影响尿路感染（UTI）。因此，将进行一项重点研究，以确定复发性尿路感染个体中的SSGP，并在尿液中寻找新的保护因子。这项研究的目的是：目标1：确定复发性UTI患者和健康对照的尿糖蛋白SSGP;目标2：建立健康个体尿液中SSGP的综合文库。 我在这些科学目标中得到了极好的研究环境的支持。波士顿儿童医院（BCH）一直被评为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有数十位国际知名的科学家，包括该项目的主要导师玛莎·摩西博士。两名科学家因在BCH的贡献而获得诺贝尔奖。重要的是，泌尿科有一个基本的、分类良好的生物样本库计划，这对本研究的成功至关重要。

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.