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: 8511347
• 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/8511347
• 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-12; Interleukin-6; Interleukins; 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. PUBLIC HEALTH RELEVANCE: Osteoarthritis (OA) is characterized by high levels of IL-12, excessive production of ROS and articular cartilage degeneration in the affected joints. However, its etiology and precise pathogenetic mechanisms remain unclear. MicroRNAs (miRNA) are a class of non-coding RNAs regulating gene expression by sequence-specific inhibition of target mRNA translation. Specific miRNAs have been shown to exhibit an altered pattern of expression in RA synovium and in other diseases. Currently there is limited information available on the expression of miRNAs in human chondrocytes but their role in OA is yet to be defined. In addition there is no information on the effects of dietary polyphenols on miRNA expression in any cell type. By studying the regulation of miRNA expression by bioavailable dietary constituents, our findings are likely to open the door for the development of novel therapeutic strategies for the treatment of OA and other diseases.

描述（由申请人提供）：骨关节炎（OA）是最常见的肌肉骨骼疾病，其特征是软骨退化和关节炎症。目前唯一有效的治疗方法是手术关节置换。 MicroRNA (miRNA) 是一类通过序列特异性抑制靶 mRNA 翻译来调节基因表达的非编码 RNA。特定的 miRNA 已被证明在 RA 滑膜和其他风湿性疾病中表现出表达模式的改变，但它们在 OA 发病机制中的作用尚未确定。在我们的初步研究中，我们分析了用 IL-12 刺激的人软骨细胞中 miRNA 的整体表达，发现功能未知的 miRNA mir-27b (mir-27b) 下调了数倍。计算机分析确定了许多可能的靶标 mRNA，包括基质金属蛋白酶-13 (MMP-13) mRNA。我们之前已经证明，标准化石榴提取物 (PE) 通过抑制体外人软骨外植体中 MMP-13 的表达以及体内小鼠模型中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对mir-27b的下调被PE阻断。我们的基本假设是“PE 通过抑制 mir-27b 和其他针对人软骨细胞中 MMP-13 mRNA 的选定 microRNA 的下调来抑制 OA 中 IL-12 诱导的软骨分解代谢效应”。这一假设的推论是“PE 的生物活性成分通过调节特定 miRNA 的表达来发挥其软骨/软骨保护作用，从而负向调节体内 MMP-13 的表达”。我们提出以下具体目标： 具体目标- 1：我们将鉴定靶向人类软骨细胞中 MMP-13 mRNA 并受 IL-12 差异调节的其他 miRNA。使用大量患者样本，我们将确认 IL-12 对 mir-27b 和上面确定的其他 miRNA 表达的影响，并确定 PE 是否在体外阻止人 OA 软骨细胞中 IL-12 的调节。具体目标 2：确定 mir-27b 和上述确定的其他 miRNA 的表达改变对体外用 IL-12 刺激的人 OA 软骨细胞中 MMP-13 表达的影响。具体目标 3：在 OA 小鼠模型中，我们将开发疾病诱导和进展期间关节中的 miRNA 表达谱。我们还将研究 PE 消耗对小鼠 OA 关节中 miR-27b 和其他靶向 MMP-13 的 miRNA 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以为开发治疗/预防 OA 的新颖且具有成本效益的治疗方法提供见解。 公众健康相关性：骨关节炎 (OA) 的特点是受影响关节中 IL-12 水平高、ROS 产生过多以及关节软骨退化。然而，其病因和确切的发病机制仍不清楚。 MicroRNA (miRNA) 是一类通过序列特异性抑制靶 mRNA 翻译来调节基因表达的非编码 RNA。特定 miRNA 已被证明在 RA 滑膜和其他疾病中表现出表达模式的改变。目前，关于人类软骨细胞中 miRNA 表达的信息有限，但它们在 OA 中的作用尚未确定。此外，没有关于膳食多酚对任何细胞类型中 miRNA 表达影响的信息。通过研究生物可利用的膳食成分对 miRNA 表达的调节，我们的研究结果可能为开发治疗 OA 和其他疾病的新治疗策略打开大门。