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

• 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
• 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

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.

项目摘要/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。 蛋白质的功能。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 强调的是先天性糖基化紊乱(CDG)，这是一组破坏 细胞糖基化机制。受影响的患者表现出严重的神经缺陷。的遗传基础 CDG提供了利用小鼠模型鉴定糖基化的神经生物学功能的机会 糖蛋白组学。了解神经系统中的糖基化将有助于阐明糖尿病的病理生理学。 CDGs和其他神经疾病，使针对糖基化途径的治疗进展成为可能，以及 告知正常的糖基化功能。 GALNT2-CDG是由GALNT2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O-糖基化的第一步。GALNT2-CDG患者患有 癫痫和全球发育迟缓。Galnt2基因敲除小鼠概括了许多患者的情况 神经缺陷。我的中心假设是神经蛋白上O-糖基化的位置特异性丢失 会导致神经功能障碍。这个项目的具体目标是找出 GALNT2-CDG的神经功能障碍。这将通过确定Galnt2的细胞起源来实现 Cre介导的神经细胞Galnt2缺失与缺乏性神经功能障碍 用糖蛋白组学鉴定这些细胞中被破坏的O-糖基化。 这项拟议的五年职业发展计划侧重于实现四个目标：培养研究技能 在小鼠模型和糖蛋白组学方面，增加了我在神经科学和糖生物学方面的知识基础， 建立一个以糖基化在神经生物学背景下的作用为重点的工作机构，并获得 过渡到独立所需的技能。导师将由公认的周兆兰博士提供 在开发和研究影响大脑的遗传性疾病小鼠模型方面处于领先地位 发育和功能，以及本杰明·加西亚博士，他是发展定量质量的公认专家 用于询问翻译后修饰的光谱技术。 通过这些调查和职业发展计划培养出来的技能将使我成为独一无二的 准备揭示CDG和其他神经疾病的糖基化介导的机制，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 工具和基础，以建立长期研究计划，以调查各种不同的病理生理学 神经系统糖化相关疾病和我准备成为独立的R01资助 内科科学家。