Validation of analytical methods for quantification of a pentasaccharide biomarker in efficacy assessment of AVV treatment for GM1 gangliosidosis

AVV 治疗 GM1 神经节苷脂沉积症疗效评估中五糖生物标志物定量分析方法的验证

• 批准号: 10360564
• 负责人: Xuntian Jiang
• 金额: $ 39.38万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10360564
• 关键词: Affect; Aftercare; Biological Assay; Biological Markers; Bone structure; Brain; Cerebrospinal Fluid; Cessation of life; Clinical; Clinical Trials; DCNU; Data; Defect; Diagnosis; Disease; Disease Progression; Evaluation; Exhibits; Felis catus; Foundations; GLB1 gene; Galactose; Ganglioside GM1; Gangliosidosis GM1; Gene therapy trial; Genetic Diseases; Glycoconjugates; Glycopeptides; Goals; Human; Impaired cognition; Keratan Sulfate; Liquid Chromatography; Liver; Longevity; Measures; Methods; Modeling; Motor; Natural History; Nerve Degeneration; Neuraxis; Neurologic Symptoms; Normal Range; Oligosaccharides; Ophthalmology; Organ; Outcome Measure; Patients; Peripheral; Pharmacodynamics; Phase I/II Clinical Trial; Phase I/II Trial; Plasma; Pre-Clinical Model; Reference Values; Reproducibility; Sampling; Sensitivity and Specificity; Spleen; Symptoms; Tissues; Treatment Effectiveness; Treatment Efficacy; Urine; Validation; Viral; Viral Genes; Visual; Work; analytical method; beta-Galactosidase; clinical efficacy; clinical predictors; disease-causing mutation; drug development; effective therapy; efficacy evaluation; enzyme activity; gene therapy; gene therapy clinical trial; longitudinal analysis; patient population; potential biomarker; premature; response; response biomarker; restoration; symptom treatment; tandem mass spectrometry; tool

PROJECT SUMMARY/ABSTRACT GM1-gangliosidosis is a rare, fatal, neurodegenerative genetic disease caused by the deficiency of β- galactosidase enzyme activity and characterized clinically by a wide range of variable neurovisceral, ophthalmological and dysmorphic features. There are currently no effective therapies for GM1-gangliosidosis and only symptomatic treatments are available. In preclinical models, adeno-associated viral (AAV) gene therapy that restores the β-galactosidase enzyme activity is the most promising therapy for delaying symptom onset, reducing storage in the brain and peripheral tissues, and increasing lifespan. These impressive results have provided the foundation for AAV gene therapy clinical trials. One of the major challenges for developing treatments for GM1-gangliosidosis is the difficulty in the evaluation of treatment efficacy due to the small and heterogeneous patient population as well as slow progression in non-infantile patients. Recently we used liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify a pentasaccharide (referred to as H3N2b) that is elevated > 20-fold in patient urine, plasma, and cerebrospinal fluid (CSF), and in the central nervous system (CNS) of the GM1-gangliosidosis cat. The CNS H3N2b levels in the GM1-gangliosidosis cat are reduced in response to AAV-treatment. H3N2b has potential as a pharmacodynamics/response biomarker for assessment of AAV-treatment efficacy in GM1-gangliosidosis. The goal of this proposal is to validate LC-MS/MS methods for determination of H3N2b in human urine, plasma, CSF, which will be used to assess AAV gene therapy treatment efficacy in a clinical trial. The aims of this application are 1) validation of LC-MS/MS methods for quantification of H3N2b in human plasma, urine, and CSF; 2) assessment of H3N2b in samples collected from GM1-gangliosidosis natural history study; and 3) application of H3N2b for assessment of treatment efficacy of AAV gene therapy. The proposed work will provide a much-needed tool for assessing therapeutic efficacy.

项目总结/摘要 GM 1-神经节苷脂沉积症是一种罕见的、致命的、神经退行性遗传性疾病，由β- 半乳糖苷酶活性和临床特征在于广泛的可变神经内脏， 眼科和畸形特征。目前还没有有效的治疗GM 1-神经节苷脂沉积症 并且只有对症治疗可用。在临床前模型中，腺相关病毒（AAV）基因治疗 恢复β-半乳糖苷酶活性是延迟症状发作的最有希望的疗法， 减少大脑和周围组织中的储存，并延长寿命。这些令人印象深刻的结果 为AAV基因治疗的临床试验提供了基础。发展的主要挑战之一是 GM 1-神经节苷脂沉积症的治疗是评价治疗效果的困难， 异质性患者群体以及非婴儿患者的缓慢进展。最近我们用液体 色谱-串联质谱法（LC-MS/MS）鉴定五糖（称为H3 N2 b） 在患者尿液、血浆和脑脊液（CSF）中以及中枢神经系统中升高> 20倍 GM 1-神经节苷脂沉积症猫的CNS H3 N2 b水平降低 对AV治疗的反应。H3 N2 b有潜力作为药效学/反应生物标志物 在GM 1-神经节苷脂沉积症中的AAV治疗功效的评估。本提案的目标是验证LC-MS/MS 建立了检测人尿液、血浆、脑脊液中H3 N2 b的方法，用于评估AAV基因 临床试验中的治疗效果。本申请的目的是1）LC-MS/MS方法的验证 用于定量人血浆、尿液和CSF中的H3 N2 b; 2）评估收集的样本中的H3 N2 b 来自GM 1-神经节苷脂沉积症自然史研究;以及3）H3 N2 b用于评估治疗功效的应用 AAV基因治疗的研究进展拟议的工作将提供一个急需的工具，用于评估治疗效果。