IgA-secreting B cell lines: a novel tool for studies of IgA nephropathy

分泌 IgA 的 B 细胞系：研究 IgA 肾病的新工具

• 批准号: 7569977
• 负责人: JAN NOVAK
• 金额: $ 18.13万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7569977
• 关键词: Acetylgalactosamine; Affect; Antibodies; Antigen-Antibody Complex; Arts; B-Lymphocytes; Basic Science; Binding; Biological Assay; Catabolism; Cell Culture Techniques; Cell Line; Cells; Characteristics; Clinical; Collaborations; Complex; Data; Defect; Deposition; Development; Diagnostic tests; Disease; Disease model; End stage renal failure; Ensure; Enzyme-Linked Immunosorbent Assay; Epitopes; Exhibits; Exposure to; Feasibility Studies; Future; Galactose; Glomerulonephritis; Glycopeptides; Goals; Human Herpesvirus 4; IgA-specific serine endopeptidase; IgA1; Immune; Immunoglobulin A; Immunoglobulin G; Individual; Injury; Kidney; Kidney Diseases; Lead; Lectin; Link; Lymphocyte; Mass Spectrum Analysis; Metabolic; Metabolic Pathway; Methods; Molecular; Molecular Analysis; Pathogenesis; Pathologic; Pathway interactions; Patients; Pattern; Peptide Hydrolases; Peripheral; Play; Polysaccharides; Principal Investigator; Quality of Care; Renal function; Resolution; Resources; Rodent Diseases; Role; Sampling; Serum; Site; Specificity; Structure; Structure of glomerular mesangium; Techniques; Testing; Therapeutic Intervention; Translating; Translations; Western Blotting; base; clinical application; clinically relevant; disorder control; enzyme linked immunospot assay; established cell line; experimental analysis; glycosylation; glycosyltransferase; immortalized cell; innovation; mesangial cell; novel; novel diagnostics; novel therapeutic intervention; novel therapeutics; programs; tool

DESCRIPTION (provided by applicant): (IgAN) is characterized by mesangial IgA1 immune deposits. These deposits originate from circulating immune complexes that contain aberrantly-glycosylated IgA1, i.e., IgA1 that is galactose-deficient in the hinge region O-linked glycans. Several lines of evidence suggest a direct causal relationship between aberrant glycosylation, the formation of immune complexes containing aberrantly glycosylated IgA1, their deposition in the mesangium, and renal injury in IgAN. These data provide the basis for the central hypothesis that an O- glycosylation abnormality in the hinge region of IgA1 is a pivotal pathogenic feature of IgAN. The ability to translate these observations into clinically relevant non-invasive diagnostic tests and novel therapeutics has been frustrated, however, by the current inability to obtain sufficient, informative samples of the pathogenic aberrantly glycosylated IgA1 for molecular analysis. We hypothesize that immortalized IgA1-producing cell lines from patients with IgAN can be used for analysis of the O-glycosylation abnormality and the specific metabolic mechanisms that result in this defect. The goal of the proposed study is to establish immortalized IgA1-secreting cell lines from circulating B cells from patients with IgAN and controls and to evaluate the potential of this new tool for advancing the studies of IgAN pathogenesis. Specifically, we propose to determine the fine structure of the O-glycosylation abnormalities of the hinge region of IgA1 secreted by the immortalized cells from IgAN patients and to identify the enzymatic pathways that may lead to this defect. The feasibility of these studies is enhanced greatly by our development of high throughput methods of analysis, state-of-the-art techniques for analysis of glycopeptides, and our extensive basic and clinical collaborations that permit testing of the hypothesis by experimental manipulation as well as clinical correlations. Relevance: IgAN is the most common primary glomerulonephritis and leads to the loss of renal function in 20-40% patients. The current inability to describe the pathogenesis of IgAN at the molecular level is a major barrier to the development of IgA-specific treatments. The proposed studies will provide an essential new tool for studies of the molecular mechanisms of pathogenesis of IgAN and a key resource for the future development and testing of potential new therapeutic interventions. This would represent a major advance in the quality of care for IgAN patients. IgAN is the most common primary glomerulonephritis. It is characterized by aberrantly- glycosylated IgA1, which represents a pivotal pathogenic feature. The proposed development of cell lines from IgAN patients will provide a significant new tool for the identification of the defects underlying the abnormal glycosylation of IgA1 in patients with IgAN and, thus, lay the basis for the translation of the basic research into clinical applications.

描述（由申请人提供）：（Igan）以肾小球IgA1免疫沉积物为特征。这些沉积物源自循环中的免疫复合物，这些免疫复合物包含异常糖基化的IgA1，即Iga1，在铰链区O-C-C-C-C-C-C-Glycans中是半乳糖缺陷的。几条证据表明，异常糖基化，含有异常的糖基化IgA1的免疫复合物的形成之间存在直接因果关系，它们在中膜中的沉积和Igan的肾损伤。这些数据为中心假设提供了基础，即IGA1铰链区域的O-糖基化异常是Igan的关键致病特征。然而，由于目前无法获得足够的，信息丰富的糖基化IGA1的病原性IGA1进行分子分析，将这些观察结果转化为临床相关的非侵入性诊断测试和新型治疗剂的能力令人沮丧。我们假设来自IGAN患者的永生化IgA1产生细胞系可用于分析O-糖基化异常和导致这种缺陷的特定代谢机制。拟议的研究的目的是建立来自Igan和对照患者循环的B细胞的永生化IgA1分泌细胞系，并评估该新工具的潜力，以推进Igan发病机理的研究。具体而言，我们建议确定来自IGAN患者的永生细胞分泌的IgA1的O-糖基化异常的精细结构，并确定可能导致这种缺陷的酶促途径。通过我们开发高通量分析方法，用于糖肽分析的最新技术以及我们广泛的基本和临床合作，可以通过实验操纵以及临床相关来检验假设，从而大大提高了这些研究的可行性。相关性：Igan是最常见的原发性肾小球肾炎，导致20-40％患者的肾功能丧失。目前无法描述分子水平上Igan的发病机理是IGA特异性治疗发展的主要障碍。拟议的研究将为研究IGAN发病机理的分子机制以及未来开发和测试潜在的新治疗干预措施的关键资源提供一个必不可少的新工具。这将代表IGAN患者护理质量的重大进步。 Igan是最常见的原发性肾小球肾炎。它的特征是异常的糖基化IgA1，它代表了一个关键的致病特征。 IGAN患者的细胞系的拟议开发将为IGA患者的IGA1异常糖基化的缺陷提供一个重要的新工具，因此为将基础研究转化为临床应用的基础是基础。