Uridylation of miRNAs by ZCCHC6 Regulates IL-6 Expression in Arthritis

ZCCHC6 对 miRNA 的尿苷化调节关节炎中 IL-6 的表达

• 批准号: 9041542
• 负责人: Tariq M Haqqi
• 金额: $ 58.04万
• 依托单位: NORTHEAST OHIO MEDICAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9041542
• 关键词: 3' Untranslated Regions; Affect; Age; American; Animal Model; Arthritis; Biological; Cartilage; Cartilage Matrix; Catabolism; Cells; Chondrocytes; Collagen; Collagen Type II; Data; Degenerative Disorder; Degenerative polyarthritis; Development; Disease Progression; Elderly; Elements; Enzymes; Gene Expression; Human; Immune; Inflammation; Inflammation Mediators; Inflammatory; Interleukin-1 beta; Interleukin-6; Joints; Knowledge; Lead; Malignant Neoplasms; Mediating; Mediator of activation protein; Messenger RNA; MicroRNAs; Modification; Mouse Strains; Musculoskeletal Diseases; Nucleotides; Occupations; Overweight; Pathogenesis; Patients; Physically Handicapped; Play; Population; Post-Transcriptional Regulation; Prevalence; Proteoglycan; Regulation; Reporting; Repression; Role; Seeds; Serum; Site; Stress; Synovial Fluid; Testing; Time; Tissues; Transferase; Translations; Untranslated RNA; Uridine; Wild Type Mouse; Woman; abstracting; articular cartilage; base; collagenase 3; cytokine; deep sequencing; design; effective therapy; interest; knock-down; loss of function; mRNA Expression; men; novel; novel marker; novel therapeutics; protein expression; public health relevance; response; targeted treatment; uridylate

 DESCRIPTION (provided by applicant): Abstract An estimated 27 million Americans age 25 and older have osteoarthritis (OA), a degenerative disease of articular cartilage characterized by loss of the cartilage matrix, mainly collagen and proteoglycans, leading to joint tissue destruction and loss of function. Before age 45, more men than women have osteoarthritis; after age 45, it is more common in women. The pro-inflammatory cytokine IL-6 has recently gained interest due to its elevated levels in synovial fluid and sera from OA patients and its capacity to upregulate the expression of MMP-13 and inhibit the expression of type-II collagen. Mature miRNAs are non-coding RNAs that inhibit the translation and stability of target mRNAs and are now known to undergo addition of non-template nucleotides (largely uridines) to the 3' ends (3'NTAs). The enzymes responsible for 3' end modifications of miRNAs have only recently been identified in cancer and immune cells but have not yet been described in cartilage or chondrocytes. Our preliminary data show that (1) ZCCHC6-a nucleotidyl transferase-is highly expressed in damaged OA cartilage; and (2) regulates IL-6 expression in human chondrocytes. ZCCHC6 is known to uridylate mRNAs thus suggesting that ZCCHC6-mediated miRNA 3'end modifications could play a significant role in the post-transcriptional regulation of IL-6, and possibly of other inflammatory mediators, in OA. We will pursue 4 specific aims to test the fundamental hypothesis that ZCCHC6 uridylates mature miRNAs in chondrocytes to regulate IL-6 expression and inflammation in OA. Specific Aim-1. Studies will test the hypothesis that "cytokine targeting miRNAs are uridylated in OA cartilage". Specific Aim-2. We will test the hypothesis that "miRNA uridylation abrogates repression of IL-6 expression in human OA chondrocytes". Specific Aim-3. We will test the hypothesis that "expression of ZCCHC6 is essential for IL-6 expression in human OA chondrocytes". Specific Aim-4. In an animal model of human OA we will test the hypothesis that "ZCCHC6 mediates miRNA uridylation and regulate IL-6 expression during the induction and progression of the disease". Knowledge gained from these studies will be important since by understanding how miRNA 3'end modifications affect the expression of inflammatory mediators in OA may lead to the development of novel biomarkers and therapies for the effective treatment of OA.

 描述（由申请人提供）：摘要估计有 2700 万 25 岁及以上的美国人患有骨关节炎 (OA)，这是一种关节软骨退行性疾病，其特征是软骨基质（主要是胶原蛋白和蛋白多糖）损失，导致关节组织破坏和关节软骨丧失。 45 岁之前，患有骨关节炎的男性多于女性；45 岁之后，由于促炎细胞因子 IL-6 引起了人们的兴趣。 OA 患者滑液和血清中其水平升高及其能力 上调 MMP-13 的表达并抑制 II 型胶原蛋白的表达 成熟 miRNA 是非编码 RNA，可抑制靶 mRNA 的翻译和稳定性，目前已知其会添加非模板核苷酸（主要是尿苷）。负责 miRNA 3' 端修饰的酶最近才在癌症和免疫细胞中被发现，但尚未在软骨或细胞中得到描述。我们的初步数据表明，(1) ZCCHC6（一种核苷酸转移酶）在受损的 OA 软骨中高表达；(2) 已知 ZCCHC6 可以调节 mRNA 的尿苷酸化，从而表明 ZCCHC6 介导的 miRNA。 3' 端修饰可能在 OA 中 IL-6 以及可能的其他炎症介质的转录后调节中发挥重要作用。我们将追求 4 个具体目标来检验 ZCCHC6 使软骨细胞中的成熟 miRNA 尿苷酸化以调节 OA 中的 IL-6 表达和炎症的基本假设。 具体目标 1 研究将检验“针对 OA 软骨中的细胞因子被尿苷酸化”的假设。具体目标 2：我们将检验“miRNA 尿苷化消除人类 OA 中 IL-6 表达的抑制”这一假设。具体目标 3。我们将检验以下假设：“ZCCHC6 的表达对于人 OA 软骨细胞中 IL-6 的表达至关重要”。具体目标 4：在人类 OA 动物模型中，我们将检验以下假设：“ ZCCHC6 在疾病的诱导和进展过程中介导 miRNA 尿苷化并调节 IL-6 表达。从这些研究中获得的知识非常重要，因为通过了解 miRNA 3' 端修饰如何影响炎症表达OA 介质可能会导致开发有效治疗 OA 的新型生物标志物和疗法。