Intersections of matrix biology with inflammation in a new model of gout

痛风新模型中基质生物学与炎症的交叉点

• 批准号: 10579760
• 负责人: Robert A. Terkeltaub
• 金额: --
• 依托单位: VA SAN DIEGO HEALTHCARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-10-01 至 2023-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10579760
• 关键词: Acceleration; Acute; Adult; Affect; Amyloid beta-Protein Precursor; Anti-Inflammatory Agents; Arthritis; Attenuated; Binding; Biological Assay; Biological Markers; Biology; C-terminal; CD44 gene; Cartilage; Cathepsin G; Chronic; Clinical Data; Cross-Sectional Studies; Crystal Formation; Crystallization; Custom; Deposition; Development; Disease; Disease Progression; Dryness; Elements; Extracellular Matrix; Female; Flare; G-substrate; Generations; Genetic; Genetic Transcription; Glycoproteins; Gout; Gouty Arthritis; Heritability; High Prevalence; Hip region structure; Homeostasis; Human; Hyperuricemia; Immunoassay; In Vitro; Inflammasome; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Arthritis; Inflammatory Response; Injectable; Injections; Joints; Knee joint; Laboratory Study; Lactoferrin; Length; Light; Link; Liquid substance; Lubricants; Macrophage; Mediating; Modeling; Mucins; Mus; Nature; Pain; Palpable; Pathogenesis; Pathway Analysis; Pathway interactions; Patients; Peptide Hydrolases; Phagocytes; Phenotype; Physiological; Prevention; Production; Proteolysis; Proteomics; Recombinants; Regulation; Resolution; Sampling; Serum; Severity of illness; Structure; Supplementation; Surface; Symptoms; Synovial Fluid; Synovitis; THBS1 gene; Testing; Therapeutic Agents; Thrombospondin 1; Thrombospondins; Tissues; Urate; Uric Acid; Validation; Variant; Veterans; Whole Blood; Work; Wrist; Xanthine Oxidase; arthropathies; articular cartilage; clinical phenotype; congenic; copolymer; disorder prevention; experimental study; foot; gain of function; gene product; glycosylation; in vivo; inhibitor; joint injury; lubricin; monocyte; new therapeutic target; novel; prevent; proband; receptor; response; screening; targeted treatment; therapeutic target; transcriptome; translational impact; treatment strategy

Hyperuricemia is fundamental in promoting the formation and accumulation of tissue deposits of monosodium urate crystals in gout, a high prevalence disease in Veterans. Urate crystals deposited in the joint promote acute, episodic flares of painful and incapacitating inflammatory arthritis by multiple pathways. Most importantly, urate crystals induce monocyte and macrophage NLRP3 inflammasome-mediated IL-1b release, and phagocyte influx and activation. Chronic inflammation lies beneath acute gout flares, including formation of articular tophi, which cause persistent synovitis and pannus-like erosive joint disease. However, several key aspects of gout pathogenesis remain unclear. In this light, asymptomatic hyperuricemia is over 5 times as prevalent than gout, and synovial fluid urate is enriched relative to serum urate in gout. Thus, passive filtering into the joint of excess circulating urate cannot be the only factor promoting tissue urate crystal deposition and symptomatic arthritis in gout. Furthermore, beyond high body urate burden, native factors that limit only ~15% of gout patients to develop palpable tophi in “tophaceous gout” are unclear. Better understanding factors that limit tophaceous gout, and their biomarkers, is vital due to association with erosive joint damage. To study novel gout prevention and therapy targets, we propose to test a paradigm-shifting core hypothesis for a gout regulation via intersection of inflammation with altered homeostasis of the extracellular matrix O-glycoprotein boundary lubricant lubricin. We specifically posit that the major gouty arthritis NLRP3/IL- 1b driver intersects with the homeostasis of lubricin to increase articular urate crystal deposition, inflammation, and disease progression in gout. Lubricin is an established, constitutive suppressor of experimental gouty inflammation. We observe that IL-1b induces xanthine oxidase and uric acid generation by macrophages, an effect blocked by exogenous lubricin. We find that lubricin markedly suppresses urate crystal formation in vitro. We also have characterized a remarkable “experiment of nature” in a 22 year old female female without hyperuricemia, who developed rapidly progressive crystal-proven gout, with classic and frequent acute flares, and erosive, destructive arthritis, affecting the feet, wrist, and hip. The proband had a heritable NLRP3 gain of function variant, an inflammatory profile of serum biomarkers and the whole blood transcriptome, attenuated serum levels of lubricin, and collective evidence consistent with altered lubricin homeostasis. Our model is that paucity of lubricin in “common gout”, due to heterogeneous mechanisms, enhances articular uric acid and increases the capacity of monosodium urate crystals to form in the joint. We thereby posit that articular lubricin deficiency, suppressible by injectable lubricin supplementation, promotes more severe acute and chronic gouty phenotypes in vivo. Translationally impactful studies will test our model by moving from in vitro to mouse to human studies. In doing so, we will test and compare mucin, and N- and C-terminal domain requirements for lubricin to limit urate crystal formation and macrophage xanthine oxidase activity, and to suppress selected gout pro-inflammatory mechanisms in macrophages. We will test the specific hypothesis that lubricin deficiency promotes ineffective resolution of IL-1b and urate crystal-induced acute articular inflammatory responses. Last, in a large, cross-sectional analysis of sera, we will test for relationships of more frequent gouty arthritis flares and tophaceous gout with low serum levels of lubricin, and certain entirely novel serum biomarkers of lubricin homeostasis pertinent to gout, including lubricin-degrading Cathepsin G activity, the Cathepsin G co-activator Lactoferrin, two inhibitors (Thrombospondin1 (TSP1/THBS1) and SERPINB6) of lubricin-degrading proteases, and the Cathepsin G substrate APP (Amyloid Precursor Protein). Completion of this work is projected to elucidate novel serum biomarkers to help guide gout treatment strategy and intensity, and new therapy targets to limit urate crystal deposition, acute and chronic synovitis, and joint damage in the disease.

高尿酸血症是促进组织沉积的形成和积累的基础 痛风中的尿酸尿酸盐晶体，这是退伍军人的高流行疾病。沉积在关节中的尿酸盐 通过多种途径促进疼痛和无能为力的炎症性关节炎的急性发作耀斑。最多 重要的是，尿酸氢酸酯诱导单核细胞和巨噬细胞NLRP3炎性体介导的IL-1B释放， 和吞噬细胞的影响和激活。慢性感染位于急性痛风之下，包括形成 关节tophi，会导致持续的滑膜炎和pannus样侵蚀性关节疾病。但是，有几个关键 痛风发病机理的各个方面尚不清楚。从此角度来看，不对称的高尿酸血症超过5倍 比痛风流行，相对于痛风中的血清尿酸酯富集。那，被动过滤 进入多余循环尿酸盐的关节 不能是促进组织尿酸盐晶体沉积和的唯一因素 痛风中有症状关节炎。此外，除了高体内尿酸伯恩，本地因素仅限于约15％ 在“托普痛风”中开发可触及的tophi的痛风患者尚不清楚。更好地理解因素 极限痛风及其生物标志物，由于与侵蚀性关节损害的关联至关重要。 为了研究新颖的预防和治疗目标，我们建议测试范式转移核心 通过炎症与细胞外稳态改变的相交痛风调节的假设 基质O-糖蛋白边界润滑剂。我们特别肯定，主要的盖蒂关节炎NLRP3/IL- 1b驱动器与润滑剂的稳态相交，以增加关节尿酸尿酸盐晶体沉积，炎症， 和痛风中的疾病进展。润滑剂是实验性痛风的构成抑制剂 炎。我们观察到IL-1B诱导巨噬细胞诱导黄嘌呤氧化物和尿酸的产生， 外源润滑剂阻断的作用。我们发现润滑剂在体外显着抑制了尿酸盐晶体形成。 我们还描述了一位22岁女性的非凡“自然实验” 高尿酸血症，他发展了迅速进行的晶体验证，并具有经典且经常急性的耀斑， 侵蚀性，破坏性关节炎，影响脚，手腕和臀部。概率具有可遗传的NLRP3收益 功能变体，血清生物标志物和整个血液转录组的炎症谱，减弱 血清润滑剂水平和与润滑剂稳态改变一致的集体证据。我们的模型是 由于异质机制，润滑剂在“普通痛风”中的稀少度增强了关节尿酸和 增加了单钠晶体在关节中形成的能力。因此，我们肯定关节润滑剂 不足，可补充注射润滑剂，可促进更严重的急性和慢性getty 体内表型。翻译具有影响力的研究将通过从体外到小鼠转移到小鼠来测试我们的模型 人类研究。在此过程中，我们将测试和比较粘蛋白以及N-和C末端域的要求 润滑素以限制尿酸氢酸酯的晶体形成和巨噬细胞黄嘌呤氧化酶活性，并抑制选定的 巨噬细胞中的痛风促炎机制。我们将测试润滑剂缺乏症的特定假设 促进IL-1B的无效分辨率，并促进尿酸盐诱导的急性关节炎症反应。最后的， 在对血清的大型横截面分析中，我们将测试更常见的盖蒂关节炎的关系 和高血清润滑剂水平低的托型gout，以及某些完全新颖的血清生物标志物的润滑剂 稳态相关的稳态，包括降解润滑剂的组织蛋白酶G活动，组织蛋白酶G共激活器 乳铁蛋白，两个抑制剂（血小板传播1（TSP1/THBS1）和SERPINB6）的溶解蛋白酶，降解蛋白酶， 和组织蛋白酶G底物应用（淀粉样蛋白前体蛋白）。这项工作的完成预计 阐明新型血清生物标志物，以帮助指导痛风治疗策略和强度，以及新的疗法靶标 限制尿酸盐晶体沉积，急性和慢性滑膜炎以及疾病中的关节损伤。