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

• 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
• 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

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.

项目摘要/摘要 糖基化的先天性疾病（CDG）是一组罕见的神经代谢遗传疾病， 破坏糖基化，将糖结构添加到蛋白质中。许多CDG基因编码研究了， 潜在的可吸毒蛋白。作为最丰富的翻译后修饰（PTM），糖基化 产生巨大的生物学变异性并介导基本的生物学过程。 CDG患者表现出来 多器官功能障碍，通常具有早期死亡率严重。 CDG的病理生理学归因于 破坏蛋白质糖基化；但是，降低糖基化蛋白的特定身份负责 大多数疾病表现尚不清楚。与理解病理机理有关的关键未满足需求 CDG患者仍然保留有效的疗法。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患者样品中糖蛋白质组学干扰的技术 可吸毒，研究的蛋白质（SLC35A2，SLC35A3和SLC39A8）。了解遗传缺陷如何 编码这些蛋白质的基因破坏糖基化将为您提供新的科学见解 CDG疾病表现的病理生理学，可能提出新的治疗策略，并将告知 糖基化的正常功能和这些研究的蛋白质。 这些选定蛋白的吸毒性在很大程度上是由于它们作为底物转运蛋白的作用，通过它 蛋白质可以吸收和将聚糖亚基的摄取和定位到内质网和高尔基体 聚糖合成和成熟的设备。大多数致病性CDG变体是肌莫尔特突变， 导致假设的假设是，转运蛋白底物的可用性增加可能会克服蛋白质功能障碍， 恢复糖基化并减轻疾病表现。除了评估糖蛋白质组学破坏 在受影响个体的标本中，该提案将测试是否补充运输蛋白底物 可以使疾病状态的糖蛋白质组学破坏归一化，并克服潜在的蛋白质功能障碍。 这项研究将使用最先进的糖蛋白质组学技术和临床相关的患者样品 阐明研究研究的CDG转运蛋白的功能。开拓仪器的进步， 糖蛋白质组学中的实验方法和计算方法可以识别 糖蛋白酶的破坏最终可实现。这些研究将确定降异性糖基化蛋白质和 患者组织中的糖材料并生成数据以解决是否补充底物的问题 在这些CDG中，可以解决疾病表现的根本原因。知识和数据 这些研究产生的将是申请资金和进行未来研究的关键 治疗由SLC35A2，SLC35A3和SLC39A8中缺陷引起的CDG疾病。