The role of microRNAs in chondrogenesis and arthritis

microRNA 在软骨形成和关节炎中的作用

• 批准号: 8265710
• 负责人: HIROSHI ASAHARA
• 金额: $ 41.02万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8265710
• 关键词: ADAMTS; Adult; Affect; Apoptosis; Appearance; Arthritis; Binding; Bioinformatics; Biological; Biological Assay; Blood Vessels; Bone Matrix; Cartilage; Cells; Cellular biology; Chondrocytes; Chondrogenesis; Collagen Type II; Degenerative polyarthritis; Development; Developmental Process; Disease; EMSA; Embryonic Development; Enzymes; Extracellular Matrix; Family; Gene Expression; Gene Targeting; Genes; Growth; HMG Domain; Homeostasis; Hypertrophy; In Vitro; Inflammatory; Joints; Knee joint; Knockout Mice; Lead; Life; Limb Development; Maintenance; Mesenchymal; Mesenchymal Stem Cells; MicroRNAs; Molecular; Molecular Target; Morphology; Movement; Mus; Nucleotides; Osteoarthrosis Deformans; Osteoblasts; Pathogenesis; Pattern; Phenotype; Physical condensation; Physiological Processes; Play; Process; Proteoglycan; Regulatory Pathway; Research; Rheumatoid Arthritis; Role; Signal Transduction; Signaling Molecule; Skeleton; Therapeutic; Therapeutic Intervention; Tissue Engineering; Tissues; Transcriptional Regulation; Transgenic Mice; Untranslated RNA; arthropathies; articular cartilage; base; bone; cartilage cell; cartilage development; early onset; extracellular; insight; interest; macromolecule; novel; novel strategies; postnatal; public health relevance; research study; scaffold; skeletal; transcription factor

DESCRIPTION (provided by applicant): The precise patterning of the developing skeletal framework relies on the appropriate control of chondrogenesis, a multistep process during which mesenchymal cells differentiate into chondrocytes. Articular cartilage destruction is a central factor in the pathogenesis of joint diseases, including osteoarthritis (OA) and rheumatoid arthritis (RA). Research on the molecular mechanisms of chondrogenesis is of developmental biological interest and has the potential to lead to novel approaches in tissue engineering and novel arthritis treatments. MicroRNAs (miRNAs) are a family of ~22-nucleotide (nt) noncoding RNAs that regulate gene expression by posttranscriptional mechanisms. To date, chondrocyte specific miRNAs, their upstream molecular signals, and target genes are largely unknown. In our preliminary experiments, we performed microarray profiling of miRNA expression in chondrocytes and miR-140 expression in cartilage development during embryogenesis and adult cartilage chondrocytes. These studies demonstrated that miR-140 has the most cartilage specific expression pattern. Cartilage specific Sox9 null mice did not express miR-140 and the potential Sox9 binding region close to pre-miR-140 was associated with Sox9 by ChIP assay and EMSA, suggesting that Sox9 is a key regulator of miR-140 gene expression. miR-140 expression was down-regulated in osteoarthritic cartilage compared with normal cartilage and by IL-1¿ stimulation of chondrocytes. miR-140 null mice showed earlier onset of osteoarthritic changes in knee joint at 6 months. Microarray and bioinformatics analysis revealed a set of possible miR-140 target genes in chondrocytes, including ADAMTS-5, which is a key tissue-degrading enzyme in osteoarthritis. These observations support the hypothesis that miR-140 is a novel regulator of cartilage development and homeostasis and those changes in its expression and function play an important role in diseases affecting articular cartilage, such as osteoarthritis. We propose the following specific aims: Aim 1: Investigate the transcriptional regulation of miR-140 expression by Sox9 in mesenchymal stem cells and chondrocytes. Aim 2: Examine the role of miR-140 in chondrogenesis and cartilage development. Aim 3: Determine the expression and function of miR-140 in osteoarthritis. Aim 4: Identify novel target genes of miR-140 in chondrocytes and mesenchymal stem cells. The proposed studies have the potential to reveal important new regulatory pathways that control cartilage development and homeostasis and open new insight on disease mechanisms and therapeutic interventions. PUBLIC HEALTH RELEVANCE: Despite substantial recent progress in understanding OA pathogenesis, disease-modifying therapies for the most prevalent joint disease are not available. Examining the role of miR-140 in MSC and chondrocytes may provide new insight into cartilage cell biology as well as a basis for novel approaches in tissue engineering and therapeutic strategies for osteoarthritis.

描述（由申请人提供）：发育中的骨骼框架的精确模式依赖于软骨形成的适当控制，软骨形成是间充质细胞分化为软骨细胞的多步骤过程。关节软骨破坏是关节疾病发病机制的核心因素，包括骨关节炎（OA）和类风湿性关节炎（RA）。对软骨形成分子机制的研究具有发育生物学意义，并有可能带来组织工程和新型关节炎治疗的新方法。 MicroRNA (miRNA) 是一个约 22 个核苷酸 (nt) 非编码 RNA 家族，通过转录后机制调节基因表达。迄今为止，软骨细胞特异性 miRNA、它们的上游分子信号和靶基因在很大程度上还是未知的。在我们的初步实验中，我们对胚胎发生和成体软骨细胞中软骨细胞中的 miRNA 表达和软骨发育中的 miR-140 表达进行了微阵列分析。这些研究表明 miR-140 具有最具软骨特异性的表达模式。软骨特异性 Sox9 缺失小鼠不表达 miR-140，并且通过 ChIP 测定和 EMSA 发现靠近 pre-miR-140 的潜在 Sox9 结合区与 Sox9 相关，表明 Sox9 是 miR-140 基因表达的关键调节因子。与正常软骨相比，通过 IL-1¿ 刺激软骨细胞，骨关节炎软骨中 miR-140 的表达下调。 miR-140 缺失小鼠在 6 个月时表现出膝关节骨关节炎变化的较早发生。微阵列和生物信息学分析揭示了软骨细胞中一组可能的 miR-140 靶基因，包括 ADAMTS-5，它是骨关节炎中的关键组织降解酶。这些观察结果支持这样的假设：miR-140是软骨发育和稳态的新型调节剂，其表达和功能的变化在影响关节软骨的疾病（例如骨关节炎）中发挥重要作用。 我们提出以下具体目标： 目标 1：研究间充质干细胞和软骨细胞中 Sox9 对 miR-140 表达的转录调控。目标 2：检查 miR-140 在软骨形成和软骨发育中的作用。目标 3：确定 miR-140 在骨关节炎中的表达和功能。目标 4：鉴定软骨细胞和间充质干细胞中 miR-140 的新靶基因。 拟议的研究有可能揭示控制软骨发育和稳态的重要新调控途径，并为疾病机制和治疗干预开辟新的见解。 公共卫生相关性：尽管最近在了解 OA 发病机制方面取得了重大进展，但针对最常见的关节疾病的疾病缓解疗法尚不可用。检查 miR-140 在 MSC 和软骨细胞中的作用可能会为软骨细胞生物学提供新的见解，并为组织工程和骨关节炎治疗策略的新方法奠定基础。