Autoantibody modulation of cartilage turnover in rheumatoid arthritis

类风湿关节炎软骨更新的自身抗体调节

• 批准号: 10199516
• 负责人: GREGG B FIELDS
• 金额: $ 42.92万
• 依托单位: FLORIDA ATLANTIC UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-09 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10199516
• 关键词: ADAMTS; Adaptive Immune System; Affect; Animal Model; Antibodies; Antigen-Presenting Cells; Apoptosis; Arthritis; Autoantibodies; Autoimmune; Autoimmune Responses; B-Lymphocytes; Binding; Cartilage; Catabolism; Chondrocytes; Citrulline; Cleaved cell; Clinical; Collagen; Collagen Type II; Digestion; Disease; Enzyme Kinetics; Epitopes; Fibroblasts; Gene Expression; Gene Proteins; Health; Hydrolysis; Immune response; Immune system; Individual; Inflammation; Inflammatory; Inhibition of Matrix Metalloproteinases Pathway; Joints; Kinetics; Light; Matrix Metalloproteinases; Minority; Onset of illness; Pathogenesis; Pathogenicity; Patients; Pattern; Physiological; Play; Population; Post-Translational Protein Processing; Process; Production; Proteins; Proteomics; Quantitative Reverse Transcriptase PCR; Regulation; Research; Research Proposals; Rheumatoid Arthritis; Rheumatoid Factor; Role; S-Phase Fraction; Signal Pathway; Site; Specificity; Structure; Swelling; Synovitis; T-Lymphocyte; Testing; Western Blotting; aggrecanase; articular cartilage; base; bone; cartilage cell; citrullinated protein; collagenase; collagenase 3; cytokine; disability; experimental study; in vivo; mouse model; novel; triple helix

ABSTRACT Rheumatoid arthritis (RA) affects 0.5-1% of the world population. RA is characterized by autoantibody production, synovial inflammation and swelling, and destruction of cartilage and bone resulting in progressive disability. It is not known whether early reactivities, such as anti-citrullinated protein antibodies (ACPAs) and rheumatoid factor (RF), are pathogenic, regulatory, or only a secondary phenomenon not related to the pathogenesis. A possible scenario is that RA occurs when the autoimmune response switches to targeting joints, in particular proteins within or adhering to cartilage. Type II collagen the major protein in joint cartilage and is also the target of most known autoantibodies that can induce arthritis. We have obtained evidence that autoimmune reactivities to type II collagen commonly occur in RA patients and may be one of the major mechanisms whereby the disease is initiated. The research plan described herein focuses on antibody modulation of type II collagen processing by matrix metalloproteinase 13 (MMP-13), the main collagenase responsible for degradation of articular cartilage during arthritis. Our hypothesis is as follows. Under normal circumstances, MMP-13 cleaves type II collagen initially at the Gly775-Leu776 bond, followed by further digestion of collagen fragments. In RA, autoantibodies to type II collagen inhibit the action of MMP-13 at different stages, resulting in the stable production of collagen fragments. The collagen fragments could activate the immune system to be more pathogenic or regulatory as well as modify chondrocyte functions, and thereby play a role in the initiation of RA. This represents a novel paradigm for RA onset and progression. To explore this hypothesis, the specific aims are to examine the effects of (1) posttranslational modification of Arg residues to citrulline on MMP-13 processing of type II collagen, (2) RA antibodies on MMP-13 processing of type II collagen and subsequent fragment production, and (3) MMP-13 derived type II collagen fragments on chondrocyte activity and in in vivo mouse models of RA. These aims will incorporate a variety of strategies, including enzyme kinetic analysis of hydrolysis of triple-helical structures, proteomics analysis of type II collagen fragments, and proliferation, qRT-PCR, FACS, and western blot analysis of chondrocytes. The present study will shed new light on the roles of specific anti- collagen antibodies in RA progression.

抽象的 类风湿关节炎（RA）影响世界人口的0.5-1％。 RA的特征是自身抗体 生产，滑膜炎症和肿胀以及软骨和骨骼的破坏，导致 渐进式残疾。尚不清楚早期重新激素，例如抗硝化蛋白抗体是否 （ACPA）和类风湿因子（RF）是致病性，调节性的，或仅是次要现象 与发病机理有关。一种可能的情况是，自动免疫响应切换时会发生RA 靶向关节，特别是内部或粘附软骨的蛋白质。 II型胶原蛋白是主要蛋白 关节软骨，也是可能诱导关节炎的最著名自身抗体的靶标。我们有 获得了证据，表明对II型胶原蛋白的自身免疫性反应率通常发生在RA患者中，并且可能 成为疾病开始的主要机制之一。本文描述的研究计划 专注于基质金属蛋白酶13（MMP-13）的II型胶原加工的抗体调节， 关节炎期间关节软骨降解的主要胶原酶。我们的假设是 跟随。在正常情况下，MMP-13裂解II型胶原蛋白最初是在Gly775-Leu776债券处 然后进一步消化胶原蛋白片段。在RA中，II型胶原蛋白的自身抗体抑制 MMP-13在不同阶段的作用，导致胶原蛋白片段的稳定产生。胶原蛋白 碎片可以激活免疫系统，以使其更具致病性或调节性并修改 软骨细胞功能，从而在RA的启动中发挥作用。这代表了一个新颖的范式 RA发作和进展。为了探讨这一假设，具体的目的是检查（1） citrulline在MMP-13加工II型胶原蛋白处理上的ARG残基的翻译后修饰，（2）RA II型胶原蛋白和随后的片段生产的MMP-13加工的抗体以及（3）MMP-13 衍生的II型胶原蛋白片段在软骨细胞活性和RA的体内小鼠模型中。这些目标 将结合各种策略，包括三螺旋水解的酶动力学分析 结构，II型胶原蛋白片段的蛋白质组学分析以及增殖，QRT-PCR，FACS和 软骨细胞的蛋白质印迹分析。本研究将为特定抗 - 的作用提供新的启示 RA进展中的胶原蛋白抗体。

项目成果

Surface Ig variable domain glycosylation affects autoantigen binding and acts as threshold for human autoreactive B cell activation.

• DOI: 10.1126/sciadv.abm1759
• 发表时间: 2022-02-11
• 期刊: Science advances
• 影响因子: 13.6
• 作者: Kissel T;Ge C;Hafkenscheid L;Kwekkeboom JC;Slot LM;Cavallari M;He Y;van Schie KA;Vergroesen RD;Kampstra ASB;Reijm S;Stoeken-Rijsbergen G;Koeleman C;Voortman LM;Heitman LH;Xu B;Pruijn GJM;Wuhrer M;Rispens T;Huizinga TWJ;Scherer HU;Reth M;Holmdahl R;Toes REM
• 通讯作者: Toes REM