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.

高尿酸血症对于促进组织沉积物的形成和积累至关重要 痛风中的单钠尿酸盐晶体，痛风是退伍军人中一种高发疾病。尿酸盐结晶沉积在关节内 通过多种途径促进疼痛和失能性炎症关节炎的急性发作。最多 重要的是，尿酸盐晶体诱导单核细胞和巨噬细胞 NLRP3 炎性体介导的 IL-1b 释放， 以及吞噬细胞的流入和激活。慢性炎症是急性痛风发作的基础，包括形成 关节痛风石，引起持续性滑膜炎和血管翳样糜烂性关节病。然而，几个关键 痛风发病机制的各个方面仍不清楚。由此看来，无症状高尿酸血症是无症状高尿酸血症的5倍以上。 比痛风更常见，并且痛风时滑液尿酸盐相对于血清尿酸盐富集。因此，无源滤波 进入过量循环尿酸盐的关节 不可能是促进组织尿酸盐晶体沉积的唯一因素 痛风症状性关节炎。此外，除了体内尿酸负担较高之外，固有因素仅限制约 15% 痛风患者中出现“痛风石性痛风”的可触及痛风石的比例尚不清楚。更好地理解影响因素 由于与糜烂性关节损伤有关，限制痛风石性痛风及其生物标志物至关重要。 为了研究新的痛风预防和治疗目标，我们建议测试范式转变的核心 通过炎症与细胞外稳态改变的交叉来调节痛风的假设 基质O-糖蛋白边界润滑剂lubricin。我们特别假设主要的痛风性关节炎 NLRP3/IL- 1b 驱动因素与润滑素的稳态相交叉，增加关节尿酸盐晶体沉积、炎症、 和痛风的疾病进展。 Lubricin 是一种已证实的实验性痛风的组成型抑制剂 炎。我们观察到 IL-1b 诱导巨噬细胞产生黄嘌呤氧化酶和尿酸，这是一种 作用被外源性润滑素阻断。我们发现润滑素在体外显着抑制尿酸盐晶体的形成。 我们还对一名 22 岁的女性进行了一项非凡的“自然实验”，但没有 高尿酸血症，患者发展为快速进展的经晶体证实的痛风，伴有典型且频繁的急性发作， 以及侵蚀性、破坏性关节炎，影响足部、手腕和臀部。先证者的可遗传 NLRP3 增益为 功能变异，血清生物标志物和全血转录组的炎症特征，减弱 润滑素的血清水平，以及与润滑素稳态改变一致的集体证据。我们的模型是这样的 由于异质机制，“普通痛风”中润滑素的缺乏会增加关节尿酸和 增加尿酸钠晶体在关节中形成的能力。因此我们假设关节润滑素 缺乏可通过注射润滑素补充剂来抑制，导致更严重的急性和慢性痛风 体内表型。具有转化影响力的研究将通过从体外研究到小鼠研究来测试我们的模型 人类研究。在此过程中，我们将测试并比较粘蛋白以及 N 端和 C 端结构域的要求 润滑素限制尿酸盐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。我们将检验润滑素缺乏的具体假设 促进 IL-1b 和尿酸盐晶体诱导的急性关节炎症反应的无效消退。最后的， 在对血清进行大型横断面分析中，我们将测试更频繁的痛风性关节炎发作之间的关系 和具有低血清润滑素水平的痛风石性痛风，以及某些全新的润滑素血清生物标志物 与痛风相关的稳态，包括润滑素降解组织蛋白酶 G 活性、组织蛋白酶 G 共激活剂 乳铁蛋白、润滑素降解蛋白酶的两种抑制剂（血小板反应蛋白 1 (TSP1/THBS1) 和 SERPINB6）， 和组织蛋白酶 G 底物 APP（淀粉样前体蛋白）。这项工作预计完成 阐明新的血清生物标志物，以帮助指导痛风治疗策略和强度以及新的治疗目标 限制尿酸盐结晶沉积、急性和慢性滑膜炎以及疾病中的关节损伤。