Suppression of MMP-13 Expression in Arthritis by Pomegranate

石榴抑制关节炎中 MMP-13 的表达

• 批准号: 8706048
• 负责人: Tariq M Haqqi
• 金额: $ 36.4万
• 依托单位: NORTHEAST OHIO MEDICAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-26 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8706048
• 关键词: ADAMTS; Affect; Arthritis; Bioavailable; Cartilage; Chondrocytes; Collagen Arthritis; Computer Simulation; Consumption; Degenerative polyarthritis; Development; Dietary Polyphenol; Dinoprostone; Disease; Disease Progression; Down-Regulation; Etiology; Exhibits; Fruit; Functional RNA; Gene Expression; Gene Targeting; Genetic Translation; Homeostasis; Human; In Vitro; Inflammatory; Inflammatory Response; Interleukin-1; Interleukin-12; Interleukin-6; Joints; Knowledge; Literature; Matrix Metalloproteinases; Messenger RNA; MicroRNAs; Modeling; Molecular; Molecular Mechanisms of Action; Molecular Profiling; Mus; Musculoskeletal Diseases; Operative Surgical Procedures; Pathogenesis; Patients; Pattern; Pomegranate; Preparation; Prevention; Principal Investigator; Production; Proteins; Regulation; Replacement Arthroplasty; Reporting; Rheumatism; Rheumatoid Arthritis; Role; Sampling; Signal Transduction; Stimulus; Symptoms; Synovial Membrane; TRAF6 gene; Testing; Therapeutic; Time; Transcript; Translations; Tumor Necrosis Factor-alpha; articular cartilage; base; cell type; collagenase 3; cost effective; cytokine; dietary constituent; effective therapy; human IRAK1 protein; in vitro Model; in vivo; insight; mouse model; novel; novel therapeutics; polyphenol; prevent; public health relevance; transmission process; treatment strategy

DESCRIPTION (provided by applicant): Osteoarthritis (OA) is the most common musculoskeletal disorder characterized by cartilage degradation and joint inflammation. Currently the only effective treatment is surgical joint replacement. MicroRNAs (miRNA) are a class of non-coding RNAs regulating gene expression by sequence specific inhibition of target mRNA translation. Specific miRNAs has been shown to exhibit altered pattern of expression in RA synovium and in other rheumatic diseases but their role in OA pathogenesis is yet to be defined. In our preliminary studies we analyzed the global expression of miRNAs in human chondrocytes stimulated with IL-12 and discovered that miRNA mir-27b (mir-27b), with no known function, was downregulated several fold. In silico analysis identified a number of possible target mRNAs including the matrix metalloproteinase-13 (MMP-13) mRNA. We have previously shown that a standardized pomegranate extract (PE) exert cartilage and chondroprotective effects by inhibiting the expression of MMP-13 in human cartilage explants in vitro and the development of inflammatory arthritis in a mouse model in vivo. Of relevance to the studies proposed in this application is our finding that downregulation of mir-27b by IL-12 was blocked by PE. Our basic hypothesis is that "PE suppresses the IL-12-induced cartilage catabolic effects in OA by inhibiting the downregulation of mir-27b and other selected microRNAs that target MMP-13 mRNA in human chondrocytes". A corollary of this hypothesis is that "bioactive constituents of PE exert their cartilage/ chondroprotective effects by modulating the expression of specific miRNAs that negatively regulate the expression of MMP-13 in vivo". We propose the following specific aims: Specific Aim- 1: We will identify other miRNAs that target MMP-13 mRNA in human chondrocytes and are differentially modulated by IL-12. Using a larger number of patient samples we will confirm the effect of IL-12 on the expression of mir-27b and other miRNAs identified above and determine whether PE prevents their modulation by IL-12 in human OA chondrocytes in vitro. Specific Aim-2: To determine the impact of altered expression of mir-27b and of other miRNAs identified above on the expression MMP-13 in human OA chondrocytes stimulated with IL-12 in vitro. Specific Aim-3: In a mouse model of OA we will develop the miRNA expression profile in the joints during disease induction and progression. We will also examine the effect of PE consumption on the expression of miR-27b and other miRNAs that target MMP-13 in mouse OA joints and correlate their expression profile with the disease progression. Knowledge gained from these studies may provide insight into developing novel and cost effective therapeutic approaches for the treatment/ prevention of OA.

描述(申请人提供)：骨关节炎(OA)是最常见的肌肉骨骼疾病，以软骨退化和关节炎症为特征。目前唯一有效的治疗方法是外科关节置换术。MicroRNAs(MiRNA)是一类非编码RNA，通过序列特异性抑制靶基因的翻译来调节基因的表达。已有研究表明，特定的miRNAs在RA滑膜和其他风湿性疾病中的表达模式发生了改变，但它们在OA发病机制中的作用尚不明确。在我们的初步研究中，我们分析了IL-12刺激的人软骨细胞中miRNAs的整体表达，发现功能未知的miRNA mir-27b(mir-27b)下调了数倍。在计算机分析中确定了一些可能的靶向mRNAs，包括基质金属蛋白酶-13(MMP13)的mRNAs。我们之前已经证明，标准化的石榴提取物(PE)在体外通过抑制人软骨外植体中基质金属蛋白酶-13的表达和在体内抑制小鼠炎症性关节炎的发展来发挥软骨和软骨保护作用。与本申请中提出的研究相关的是，我们发现IL-12下调mir-27b的作用可被PE阻断。我们的基本假设是“PE通过抑制mir-27b和其他靶向人软骨细胞中的基质金属蛋白酶-13mRNAs的下调，从而抑制IL-12诱导的骨性关节炎的软骨分解代谢效应”。这一假说的一个推论是“PE的生物活性成分通过调节体内特定的miRNAs的表达来发挥其软骨/软骨保护作用，该miRNAs负向调节MMP13的表达”。我们提出了以下特定的目标：特定的目标-1：我们将确定其他针对人软骨细胞中的基质金属蛋白酶-13mRNA的miRNAs，并受到IL-12的差异调控。利用大量的患者样本，我们将证实IL-12对mir-27b和其他miRNAs表达的影响，并确定PE是否能在体外阻止IL-12对人骨关节炎软骨细胞的调节。特异性目的-2：确定mir-27b和其他miRNAs的表达变化对IL-12刺激的人骨性关节炎软骨细胞中基质金属蛋白酶-13表达的影响。具体目标-3：在小鼠骨性关节炎模型中，我们将建立疾病诱导和进展过程中关节中miRNA的表达谱。我们还将检测PE摄入对小鼠骨关节炎关节中miR-27b和其他针对基质金属蛋白酶-13的miRNAs表达的影响，并将它们的表达谱与疾病进展相关联。从这些研究中获得的知识可能为开发治疗/预防骨性关节炎的新的、具有成本效益的治疗方法提供洞察力。

项目成果

Effect of a Herbal-Leucine mix on the IL-1β-induced cartilage degradation and inflammatory gene expression in human chondrocytes.

• DOI: 10.1186/1472-6882-11-66
• 发表时间: 2011-08-19
• 期刊: BMC complementary and alternative medicine
• 作者: Akhtar N;Miller MJ;Haqqi TM
• 通讯作者: Haqqi TM

Pomegranate extract inhibits the interleukin-1β-induced activation of MKK-3, p38α-MAPK and transcription factor RUNX-2 in human osteoarthritis chondrocytes.

• DOI: 10.1186/ar3166
• 发表时间: 2010
• 期刊: Arthritis research & therapy
• 影响因子: 4.9
• 作者: Rasheed Z;Akhtar N;Haqqi TM
• 通讯作者: Haqqi TM

Epigallocatechin-3-gallate suppresses the global interleukin-1beta-induced inflammatory response in human chondrocytes.

• DOI: 10.1186/ar3368
• 发表时间: 2011-06-17
• 期刊: Arthritis research & therapy
• 影响因子: 4.9
• 作者: Akhtar N;Haqqi TM
• 通讯作者: Haqqi TM