Project 2: GeorgiaCenter for the Investigation of Factor VIII Inhibitors and Glycosylation

项目 2：乔治亚因子 VIII 抑制剂和糖基化研究中心

• 批准号: 10227917
• 负责人: Li Lei
• 金额: $ 30.89万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10227917
• 关键词: Affect; Amino Acids; Antibodies; Attention; B-Lymphocytes; Birth; Blood Coagulation Disorders; Carbohydrates; Cells; Clinical; Clinical Treatment; Coagulants; Coagulation Process; Complex; Development; Diagnosis; Environment; Epitopes; F8 gene; Factor V; Factor VIII; Functional disorder; Glycopeptides; Glycoproteins; Health Care Costs; Hemophilia A; Immune Tolerance; Immune system; Immunoglobulin G; Immunoglobulins; Intravenous; Investigation; Link; Measurement; Medicine; Modification; Morbidity - disease rate; Patient Monitoring; Patients; Pattern; Perception; Plasma; Property; Protein Glycosylation; Proteins; Recombinants; Replacement Therapy; Reporting; Research; Risk; Sampling; Surrogate Markers; Surveys; T-Lymphocyte; Testing; Translations; Variant; base; de novo mutation; glycosylation; immunogenic; immunogenicity; inhibitor/antagonist; innovation; male; neutralizing antibody; novel; protein aggregation; tool

Summary Congenital Hemophilia A (HA) is an X-linked bleeding disorder due to a lack or dysfunction of coagulation FVIII (FVIII). Nearly 1 in 5,000 live male births are diagnosed with HA with 40% of whom involving of sporadic cases caused by de novo mutations. Plasma-derived FVIII (pdFVIII) and recombinant FVIII (rFVIII) are two major intravenous replacement therapy medicines for HA treatment. FVIII neutralizing antibodies, or inhibitors, occur in 25-30% of severe HA patients receiving protein replacement. These FVIII inhibitors essentially abolish the efficacy of the coagulant treatment, resulting in a considerable increase in the morbidity and healthcare cost. Although both rFVIII and pdFVIII share similar to identical primary amino acids, the risk of FVIII antibodies in previously untreated patients (PUPs) was reported to be higher following rFVIII treatment. As a glycoprotein, pdFVIII and rFVIII are different in glycosylation, a post-translation modification (PTM) that is capable of alter the immunogenicity of protein Therefore, in this project, we will develop a set of novel glycoanalysis toolset for FVIII inhibitors. In order to investigate the effect of the host glyco environment on FVIII, we propose to use FVIII closely associated VWF and FV as surrogate marker to show to potential effect on FVIII glycosylation in HA patients who developed FVIII inhibitors. Also, based on our developed MS-based approach, glycosylation pattern of total immunoglobulins, specific FVIII inhibitors, FV, VWF, and overall glycome changes of the host cell environment will be thoroughly characterized in a large number of HA patients’ samples, close attention to those undergone immune tolerance induction (ITI). The overall results obtained from this project will be significant for understanding the fundamental mechanism of FVIII immunogenicity.

概括 先天性血友病A（HA）是由于缺乏或功能障碍FVIII而导致的X连锁出血障碍 （FVIII）。近5,000名活着的男性出生中有近1个被诊断出患有HA，其中40％涉及零星病例 由从头突变引起。血浆衍生的FVIII（PDFVIII）和重组FVIII（RFVIII）是两个主要 用于HA治疗的静脉替代治疗药物。 FVIII中和抗体或抑制剂发生 在25-30％的严重HA患者接受蛋白质替代的患者中。这些FVIII抑制剂基本上废除了 凝结治疗的功效，导致发病率和医疗保健成本的考虑增加。 尽管RFVIII和PDFVIII均具有相似的原代氨基酸，但FVIII抗体的风险 据报道，在RFVIII治疗后，以前未经治疗的患者（幼崽）较高。作为糖蛋白， PDFVIII和RFVIII的糖基化是不同的，糖基化是一种能够改变的翻译后修饰（PTM） 蛋白质的免疫原性 因此，在这个项目中，我们将开发一套针对FVIII抑制剂的新型甘油分析工具集。为了 调查宿主Glyco环境对FVIII的影响，我们建议使用FVIII密切相关的VWF 和FV作为替代标记，以表现出对患有FVIII的HA患者的FVIII糖基化的潜在影响 抑制剂。同样，基于我们开发的基于MS的方法，总免疫球蛋白的糖基化模式， 特定的FVIII抑制剂，FV，VWF和宿主细胞环境的总体变化将彻底进行 以大量HA患者的样本为特征，密切关注那些受到免疫耐受性的人 归纳（ITI）。从该项目获得的总体结果对于理解基本 FVIII免疫原性的机制。