Identifying understudied protein-related glycoproteome disruption in Congenital Disorders of Glycosylation

识别先天性糖基化障碍中尚未研究的蛋白质相关糖蛋白组破坏

基本信息

批准号：
10725869
负责人：
Andrew Charles Edmondson
金额：
$ 18.8万
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：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10725869
关键词：
Address Affect Applications Grants Biological Biological Process Carrier Proteins Cataloging Cells Clinical Clinical Research Congenital disorders of glycosylation Data Defect Disease Endoplasmic Reticulum Exhibits Fibroblasts Functional disorder Funding Future Genes Genetic Diseases Glycopeptides Glycoproteins Goals Golgi Apparatus Health Human Individual Knowledge Lectin Life Expectancy Link Live Birth Mass Spectrum Analysis Mediating Methodology Methods Mutation N-Glycosylation Site Nature Organ Pathogenicity Pathway interactions Patients Peptides Phase III Clinical Trials Phenotype Physicians Polysaccharides Post Translational Modification Analysis Post-Translational Protein Processing Protein Glycosylation Proteins Proteome Proteomics Quality of life Rare Diseases Research Personnel Role Sampling Scientist Specimen Structure Supplementation Symptoms System Technology Testing Therapeutic Tissues United States National Institutes of Health Variant clinically relevant effective therapy efficacy evaluation frontier glycoproteomics glycosylation improved insight instrumentation mortality novel novel therapeutics protein protein interaction proteogenomics rare genetic disorder sugar treatment strategy uptake

项目摘要

PROJECT SUMMARY/ABSTRACT The congenital disorders of glycosylation (CDG) are a group of rare neurometabolic genetic diseases that disrupt glycosylation, the addition of sugar structures to proteins. Many CDG genes encode understudied and potentially druggable proteins. As the most abundant post-translational modification (PTM), glycosylation generates immense biological variability and mediates fundamental biological processes. CDG patients exhibit multiorgan dysfunction, which is often severe with early mortality. Pathophysiology of CDG is attributed to disrupted protein glycosylation; however, the specific identities of hypoglycosylated proteins responsible for most disease manifestations are unknown. Critical unmet needs related to understanding pathomechanisms and effective therapies remain for CDG patients. Here, we propose to utilize mass spectrometry-based technologies to evaluate glycoproteomic disruptions in CDG patient samples with pathogenic variants in druggable, understudied proteins (SLC35A2, SLC35A3, and SLC39A8). Understanding how genetic defects in genes encoding these proteins disrupts glycosylation will provide new scientific insights into the pathophysiology of CDG disease manifestations, may suggest novel treatment strategies, and will inform normal function of glycosylation and these understudied proteins. The druggability of these selected proteins is largely due to their roles as substrate transporters, through which the proteins enable uptake and localization of glycan subunits to the endoplasmic reticulum and Golgi apparatus for glycan synthesis and maturation. Most pathogenic CDG variants are hypomorphic mutations, leading to the hypothesis that increased availability of transporter substrate may overcome protein dysfunction, restore glycosylation, and mitigate disease manifestations. In addition to evaluating glycoproteomic disruption in specimens from affected individuals, this proposal will test whether transporter substrate supplementation can normalize the glycoproteomic disruption of the disease state and overcome underlying protein dysfunction. This study will use using state-of-the-art glycoproteomics technologies and clinically relevant patient samples to elucidate functions of understudied CDG transporter proteins. Pioneering advances in instrumentation, experimental methodologies, and computational approaches in glycoproteomics make identification of glycoproteome disruption finally achievable. These studies will identify hypoglycosylated proteins and glycosites in patient tissues and generate data to address the question of whether substrate supplementation in these CDG may address the underlying causes of disease manifestations. The knowledge and data generated from these studies will be pivotal for applying for funding and carrying out future studies aimed at treating CDG disorders caused by defects in SLC35A2, SLC35A3, and SLC39A8.

项目概要/摘要先天性糖基化障碍（CDG）是一组罕见的神经代谢遗传病，破坏糖基化，即向蛋白质添加糖结构。许多 CDG 基因编码尚未被研究和潜在可药物化的蛋白质。作为最丰富的翻译后修饰（PTM），糖基化产生巨大的生物变异性并介导基本的生物过程。 CDG 患者表现多器官功能障碍，通常很严重并导致早期死亡。 CDG 的病理生理学归因于蛋白质糖基化被破坏；然而，低糖基化蛋白质的具体身份负责大多数疾病的表现是未知的。与了解病理机制相关的关键未满足需求 CDG 患者仍有有效的治疗方法。在这里，我们建议利用基于质谱的评估具有致病性变异的 CDG 患者样本中糖蛋白组破坏的技术可药物化、正在研究的蛋白质（SLC35A2、SLC35A3 和 SLC39A8）。了解遗传缺陷如何影响编码这些蛋白质的基因会破坏糖基化，这将为我们提供新的科学见解。 CDG 疾病表现的病理生理学，可能会提出新的治疗策略，并将提供信息糖基化的正常功能和这些待研究的蛋白质。这些选定蛋白质的成药性很大程度上是由于它们作为底物转运蛋白的作用，通过它这些蛋白质能够将聚糖亚基摄取并定位到内质网和高尔基体用于聚糖合成和成熟的装置。大多数致病性 CDG 变异是亚等位性突变，导致这样的假设：增加转运底物的可用性可以克服蛋白质功能障碍，恢复糖基化并减轻疾病表现。除了评估糖蛋白组破坏之外在受影响个体的样本中，该提案将测试是否补充转运蛋白底物可以使疾病状态的糖蛋白组破坏正常化并克服潜在的蛋白质功能障碍。该研究将使用最先进的糖蛋白组学技术和临床相关的患者样本阐明正在研究的 CDG 转运蛋白的功能。仪器仪表领域的开创性进步，糖蛋白质组学中的实验方法和计算方法可以识别糖蛋白组破坏终于可以实现。这些研究将鉴定低糖基化蛋白质和患者组织中的糖位点并生成数据以解决是否补充底物的问题这些 CDG 可能会解决疾病表现的根本原因。知识和数据这些研究产生的结果对于申请资金和开展未来的研究至关重要治疗由 SLC35A2、SLC35A3 和 SLC39A8 缺陷引起的 CDG 疾病。