RA synovial fibroblast exosomes(RA-EXo) mediated bone erosion via AhR/TRAF2pathway

RA滑膜成纤维细胞外泌体（RA-EXo）通过AhR/TRAF2通路介导骨侵蚀

• 批准号: 10622327
• 负责人: HUANG-GE ZHANG
• 金额: --
• 依托单位: LOUISVILLE VA MEDICAL MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10622327
• 关键词: ARA9 protein; Affect; Animal Model; Arthritis; Aryl Hydrocarbon Receptor; Attenuated; Autoimmune Diseases; Biological; Biological Markers; Biological Response Modifiers; Biotin; Biotinylation; Bone Development; CRISPR/Cas technology; Cartilage; Cells; Chondrocytes; Chronic; Complex; Cytosol; Data; Development; Diagnosis; Dissociation; Endothelial Cells; Exocytosis; Extracellular Space; Fibroblasts; Future; Guidelines; Health Status; Heat-Shock Proteins 90; High Prevalence; Human; Hyperplasia; Immune; Immunocompetent; Implant; Individual; Inflammation; Invaded; Knock-out; Knowledge; Label; Ligase; MAPK8 gene; Mediating; Mediator; Military Personnel; Modeling; Molecular Target; Mus; Mutate; NF-kappa B; Osteitis; Outcome; Outcome Study; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Pharmacotherapy; Phosphorylation; Phosphotransferases; Play; Population; Predictive Value; Prevention; Process; Proliferating; Proteins; Publishing; Receptor Activation; Research; Rheumatoid Arthritis; Role; SCID Mice; Severities; Site; Sorting; Synovial Cell; Synovial Fluid; Synovial Membrane; System; TNF Receptor-Associated Factors; TNF gene; TNF receptor-associated factor 2; Testing; Therapeutic; Tissues; United States Department of Veterans Affairs; Vesicle; Veterans; Virulence Factors; Woman; arthritis therapy; arthropathies; bone; bone erosion; cartilage degradation; cell type; disability; exosome; experimental study; extracellular; improved; in vivo; intercellular communication; joint destruction; joint inflammation; joint injury; member; men; mouse model; novel therapeutics; patient population; personalized medicine; precision drugs; predictive marker; prevent; protein protein interaction; receptor expression; recruit; valosin-containing protein

Rheumatoid Arthritis (RA) is more prevalent among veterans than nonveterans. Women comprise an increasing proportion of military personnel and have a higher prevalence of arthritis than men. Thus, RA is a highly relevant problem for the Veterans Administration (VA). In RA synovium, rheumatoid arthritis synovial fibroblasts (RASFs) are hyper proliferated, the leading cell type in the terminal layer of the hyperplastic synovial tissue that invades and degrades adjacent cartilage and bone. TRAF2 mediated activation of NF-B and JNK leads to hyperproliferation of RASFs, chronic inflammation and the erosive arthritis which is the hallmark of RA. How NF- B and JNK is activated in rheumatoid synovial fibroblast remains elusive, though studies have demonstrated that the blockade of TNF mediated activation of NF-B and JNK attenuates the RA progression. However, a large population of RA patients are non-response to anti-TNF therapy. Research from our group and others has shown that exosomes released from rheumatoid synovial fibroblasts (RA-Exo) contribute to RA pathogenicity and inflammation in several different animal models of joint disease, including RA. Exosomes are small cell derived vesicles that are 30-100 nm in size and of endocytic origin. Exosomes released by exocytosis into the extracellular space, where they are considered important drivers of intercellular communication. Exosomes in synovial fluid of RA patients can lead to inflammation, degeneration of cartilage, and destruction of joints. However, exosomes in synovial fluid could be released by different types of cells including RASFs in the synovium. Which cell types release exosomes that contribute to joint inflammation and degeneration of cartilage is not well studied. Of particular relevance to this proposal, we have demonstrated that RASF exosomes (RA-Exo) activate the TNF- receptor-associated factor 2 (TRAF2) mediated activation of the NF-B and JNK pathways is promoted by stabilization of TRAF2. More importantly, our preliminary data clearly demonstrate that aryl hydrocarbon receptor (AhR) expression is significantly elevated in synovium from patients with RA. Expression positively correlates with bone erosion and is localized with TRAF2, strongly supporting the relevance of this pathway in the pathogenesis of RA in humans. Mechanistically, AhR is enriched in the RA-Exo and RA-Exo AhR is required for stabilizing TRAF2. Our OVERALL HYPOTHESIS is that targeting AhR centered networks in RA-Exo leads to TRAF2 degradation, and the prevention of TRAF2 mediated activation of the NF-B and JNK pathways in RASFs. Therefore, disruption of the AhR network in RA exosomes will eliminate/attenuate bone erosion and inflammation by affecting RA-Exo mediated pathways contributing to the development of bone erosion in synovium of RA patients. Our plan to accomplish this objective are outlined in these three specific aims: 1. Identify the RA-Exo AhR network that results in the development of arthritis in the mouse models. 2. Determine whether RA-Exo AhR-associated kinase activity mediates dissociation of TRAF2AF from the TRAF2 complex. 3. Determine whether blocking the recruitment of VCIP135 to RA-Exo AhR results in preventing the development of erosive arthritis in the huRASF/SCID mouse model. The positive outcome of this study for future products would provide a basis for developing mechanism-driven novel drugs for RA therapy. Moreover, it is of high value for predicting biomarkers from RA-Exo of non-responsive individuals prior to commencing anti-TNF or personalized drug therapy.

类风湿性关节炎 (RA) 在退伍军人中比非退伍军人更常见。妇女包括 军人比例不断增加，关节炎患病率高于男性。因此，RA 是一种高度 退伍军人管理局（VA）的相关问题。在 RA 滑膜中，类风湿性关节炎滑膜成纤维细胞 (RASF) 过度增殖，是增生性滑膜组织终末层的主要细胞类型， 侵入并降解邻近的软骨和骨骼。 TRAF2 介导的 NF-κB 和 JNK 激活导致 RASF 过度增殖、慢性炎症和糜烂性关节炎（RA 的标志）。如何 NF- 尽管研究表明，B 和 JNK 在类风湿滑膜成纤维细胞中被激活仍然难以捉摸 阻断 TNFα 介导的 NF-κB 和 JNK 激活可减弱 RA 的进展。然而，一个大 RA 患者群体对抗 TNFα 治疗无反应。我们小组和其他人的研究表明 类风湿滑膜成纤维细胞释放的外泌体 (RA-Exo) 有助于 RA 致病性， 几种不同的关节疾病动物模型（包括 RA）中的炎症。外泌体源自小细胞 大小为 30-100 nm 且具有内吞起源的囊泡。外泌体通过胞吐作用释放到细胞外 空间，它们被认为是细胞间通讯的重要驱动因素。滑液中的外泌体 RA患者可导致炎症、软骨退化和关节破坏。然而，外泌体在 滑液可以由不同类型的细胞释放，包括滑膜中的 RASF。哪些细胞类型 释放导致关节炎症和软骨变性的外泌体尚未得到充分研究。 与该提案特别相关的是，我们已经证明 RASF 外泌体 (RA-Exo) 激活 TNF- 受体相关因子 2 (TRAF2) 介导的 NF-κB 和 JNK 通路的激活由 TRAF2 的稳定性。更重要的是，我们的初步数据清楚地表明芳烃受体 (AhR) 表达在 RA 患者的滑膜中显着升高。表达量正相关 骨侵蚀并且与 TRAF2 相关，有力地支持了该通路在发病机制中的相关性 人类 RA 的发生。从机制上讲，AhR 在 RA-Exo 中富集，并且 RA-Exo AhR 是稳定 交易2。我们的总体假设是，针对 RA-Exo 中以 AhR 为中心的网络会导致 TRAF2 降解，以及防止 TRAF2 介导的 RASF 中 NF-κB 和 JNK 通路的激活。 因此，破坏 RA 外泌体中的 AhR 网络将消除/减轻骨侵蚀和炎症 通过影响 RA-Exo 介导的途径，促进 RA 滑膜骨侵蚀的发展 患者。 我们实现这一目标的计划概述为以下三个具体目标： 1. 鉴定导致小鼠模型发生关节炎的 RA-Exo AhR 网络。 2.确定RA-Exo AhR相关激酶活性是否介导TRAF2AF解离 来自 TRAF2 复合体。 3. 确定阻断 VCIP135 向 RA-Exo AhR 的募集是否可以防止 huRASF/SCID 小鼠模型中糜烂性关节炎的发展。 这项研究对未来产品的积极成果将为开发机制驱动的新型产品提供基础 用于 RA 治疗的药物。此外，它对于预测无反应性 RA-Exo 的生物标志物具有很高的价值。 在开始抗 TNFα 或个性化药物治疗之前。