O-glycosylation mechanisms of neurological deficits in congenital disorders of glycosylation

先天性糖基化障碍神经功能缺损的O-糖基化机制

基本信息

批准号：
10040788
负责人：
Andrew Charles Edmondson
金额：
$ 18.08万
依托单位：
CHILDREN'S HOSP OF PHILADELPHIA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10040788
关键词：
Affect Astrocytes Biology Brain Career Mobility Cell Line Cells Cellular Morphology Complex Congenital disorders of glycosylation Constitutional Development Development Plans Developmental Delay Disorders Disease Epilepsy Event Exhibits Family member Foundations Functional disorder Funding Future Genetic Genetic Diseases Glycobiology Glycoproteins Goals Human Individual International Investigation Knockout Mice LoxP-flanked allele Mass Spectrum Analysis Mediating Mentors Microcephaly Modeling Molecular Morphology Mucins Mus Mutation Nervous System Physiology Nervous system structure Neurobiology Neurologic Neurologic Deficit Neurologic Dysfunctions Neurons Neurosciences Pathway interactions Patients Phenotype Physicians Post-Translational Protein Processing Process Protein Isoforms Proteins Rare Diseases Research Role Scientist Signal Transduction Site Symptoms Techniques Therapeutic Training behavioral phenotyping career development cell motility cell type comparative conditional knockout design glycoproteomics glycosylation glycosyltransferase in vitro Assay insight knowledge base molecular phenotype motor deficit mouse model nervous system disorder neurotransmitter release programs protein function rare genetic disorder relating to nervous system sensory integration skills tool

项目摘要

PROJECT SUMMARY/ABSTRACT Glycosylation is an essential, post-translational modification with complex and poorly understood roles in protein function. My long-term objective is to elucidate the neurobiological functions of glycosylation, including identifying the roles of critical glycosylation sites in neuronal protein function. The importance of glycosylation is emphasized by the congenital disorders of glycosylation (CDG), a group of genetic disorders that disrupt cellular glycosylation machinery. Affected patients exhibit severe neurological deficits. The genetic basis of CDG provides an opportunity to identify the neurobiological functions of glycosylation using mouse models and glycoproteomics. Understanding glycosylation in the nervous system will elucidate the pathophysiology of CDGs and other neurological diseases, enable therapeutic advances targeting glycosylation pathways, and inform normal function of glycosylation. GALNT2-CDG is a new CDG type caused by biallelic mutations in GALNT2, which encodes a critical glycosyltransferase initiating the first step in mucin-type O-glycosylation. GALNT2-CDG patients suffer from epilepsy and global developmental delay. Galnt2 constitutional knock-out mice recapitulate many of the patient neurological deficits. My central hypothesis is that site-specific loss of O-glycosylation on neural proteins contributes to neurological dysfunction. The specific objective of this project is to identify the cause of neurological dysfunction in GALNT2-CDG. This will be achieved by determining cellular origins of Galnt2 deficiency-mediated neurological deficits using Cre-mediated deletion of Galnt2 in neural cells and by identifying disrupted O-glycosylation in these cells using glycoproteomics. This proposed five-year career development plan focuses on achieving four objectives: develop research skills in mouse models and glycoproteomics, increase my knowledgebase in neuroscience and glycobiology, establish a body of work focusing on the role of glycosylation in the context of neurobiology, and obtain the necessary skills to transition to independence. Mentoring will be provided by Dr. Zhaolan Zhou, a recognized leader in the development and investigation of mouse models of genetic disorders that affect brain development and function, and Dr. Benjamin Garcia, a recognized expert in developing quantitative mass spectrometry techniques to interrogate post-translational modifications. The skill set developed through these investigations and career development plan will make me uniquely poised to uncover glycosylation-mediated mechanisms of CDG and other neurological diseases, as well as to elucidate the critical roles of glycosylation in human neurological function. These studies will generate new tools and a foundation to establish a long-term research program to investigate the pathophysiology of various glycosylation-related disorders in the nervous system and prepare me to become an independent R01-funded physician scientist.

项目概要/摘要糖基化是一种重要的翻译后修饰，其作用复杂且知之甚少。蛋白质功能。我的长期目标是阐明糖基化的神经生物学功能，包括确定关键糖基化位点在神经元蛋白质功能中的作用。糖基化的重要性先天性糖基化障碍（CDG）强调了这一点，这是一组破坏糖基化的遗传性疾病细胞糖基化机器。受影响的患者表现出严重的神经功能缺损。遗传基础 CDG 提供了使用小鼠模型识别糖基化的神经生物学功能的机会糖蛋白质组学。了解神经系统中的糖基化将阐明其病理生理学 CDG 和其他神经系统疾病使针对糖基化途径的治疗取得进展，并且告知糖基化的正常功能。 GALNT2-CDG 是由 GALNT2 的双等位基因突变引起的一种新的 CDG 类型，编码一个关键的糖基转移酶启动粘蛋白型 O-糖基化的第一步。 GALNT2-CDG 患者患有癫痫和整体发育迟缓。 Galnt2 体质敲除小鼠再现了许多患者的情况神经功能缺陷。我的中心假设是神经蛋白上 O-糖基化的位点特异性丢失导致神经功能障碍。该项目的具体目标是确定问题的原因 GALNT2-CDG 的神经功能障碍。这将通过确定 Galnt2 的细胞起源来实现使用 Cre 介导的神经细胞中 Galnt2 的缺失来治疗缺陷介导的神经功能缺陷使用糖蛋白质组学鉴定这些细胞中被破坏的 O-糖基化。拟议的五年职业发展计划重点关注实现四个目标：培养研究技能在小鼠模型和糖蛋白组学方面，增加我在神经科学和糖生物学方面的知识库，建立一个专注于糖基化在神经生物学中的作用的工作体系，并获得过渡到独立的必要技能。周兆兰博士将提供指导，他是一位公认的影响大脑的遗传性疾病小鼠模型开发和研究的领导者发育和功能，以及本杰明·加西亚博士（Benjamin Garcia），一位公认的定量质量开发专家光谱测定技术来询问翻译后修饰。通过这些调查和职业发展计划培养的技能将使我独一无二准备揭示 CDG 和其他神经系统疾病的糖基化介导机制，以及阐明糖基化在人类神经功能中的关键作用。这些研究将产生新的建立长期研究计划以调查各种疾病病理生理学的工具和基础神经系统中与糖基化相关的疾病，让我做好成为独立 R01 资助的人的准备医师科学家。