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 类风湿性关节炎 (RA) 影响着世界人口的 0.5-1%。 RA的特征是自身抗体 产生，滑膜炎症和肿胀，以及软骨和骨的破坏，导致 进行性残疾。目前尚不清楚是否存在早期反应，例如抗瓜氨酸蛋白抗体 (ACPA) 和类风湿因子 (RF) 是致病性的、调节性的，或者只是一种次要现象，而不是 与发病机制有关。一种可能的情况是，当自身免疫反应发生切换时，就会发生 RA 靶向关节，特别是软骨内或粘附于软骨的蛋白质。 II 型胶原蛋白是体内的主要蛋白质 关节软骨，也是大多数已知的可诱发关节炎的自身抗体的目标。我们有 获得的证据表明，对 II 型胶原蛋白的自身免疫反应常见于 RA 患者，并且可能 是引发疾病的主要机制之一。本文描述的研究计划 专注于基质金属蛋白酶 13 (MMP-13) 对 II 型胶原加工的抗体调节， 负责关节炎期间关节软骨降解的主要胶原酶。我们的假设是 接下来。正常情况下，MMP-13 首先在 Gly775-Leu776 键处裂解 II 型胶原， 然后进一步消化胶原蛋白碎片。在 RA 中，II 型胶原蛋白的自身抗体会抑制 MMP-13在不同阶段的作用，导致胶原片段的稳定产生。胶原蛋白 片段可以激活免疫系统，使其更具致病性或调节性以及修饰 软骨细胞功能，从而在 RA 的引发中发挥作用。这代表了一个新颖的范例 RA 的发病和进展。为了探索这一假设，具体目标是检验 (1) 的影响 在 II 型胶原的 MMP-13 加工过程中，Arg 残基翻译后修饰为瓜氨酸，(2) RA 影响 II 型胶原 MMP-13 加工和随后片段生成的抗体，以及 (3) MMP-13 衍生的 II 型胶原片段对软骨细胞活性和 RA 体内小鼠模型的影响。这些目标 将结合多种策略，包括三螺旋水解的酶动力学分析 结构、II 型胶原片段的蛋白质组学分析和增殖、qRT-PCR、FACS 和 软骨细胞的蛋白质印迹分析。本研究将为特定抗- RA 进展中的胶原抗体。

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