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（由适用提供）：发育中的骨骼框架的精确模式依赖于适当的软骨形成，这是一种多步骤过程，在此过程中，间充质细胞分化为软骨细胞。关节软骨破坏是关节疾病发病机理的核心因素，包括骨关节炎（OA）和类风湿关节炎（RA）。关于软骨发生的分子机制的研究具有发育生物学的兴趣，并有可能导致组织工程和新型关节炎治疗的新方法。 microRNA（miRNA）是〜22-核苷酸（NT）非编码RNA的家族，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尚不清楚。在我们的初步实验中，我们在胚胎发生和成人软骨软骨细胞过程中对软骨细胞中miRNA表达和miR-140表达进行了微阵列分析。这些研究表明，miR-140具有最多的软骨特异性表达模式。软骨特异性Sox9 null小鼠未表达miR-140，而接近MIR-140的潜在SOX9结合区域与ChIP Assay和EMSA与SOX9相关，这表明SOX9是miR-140基因表达的关键调节剂。与正常软骨和通过IL-1刺激软骨细胞相比，在骨关节炎软骨中，miR-140表达在骨关节炎软骨中下调。 miR-140零小鼠早先显示了6个月时膝关节骨关节炎的变化。微阵列和生物信息学分析揭示了包括ADAMTS-5在内的软骨细胞中的一组可能的miR-140靶基因，这是骨关节炎的关键组织降解酶。这些观察结果支持以下假设：miR-140是软骨发育和稳态的新调节剂，其表达和功能的变化在影响关节软骨的疾病中起着重要作用，例如骨关节炎。我们提出以下特定目的：目标1：研究间充质干细胞和软骨细胞中SOX9对miR-140表达的转录调节。目标2：检查miR-140在软骨发生和软骨发育中的作用。 AIM 3：确定miR-140在骨关节炎中的表达和功能。 AIM 4：确定软骨细胞和间充质干细胞中miR-140的新型靶基因。拟议的研究有可能揭示重要的新调节途径，以控制软骨发展和体内平衡，并开放有关疾病机制和治疗干预措施的新见解。 公共卫生相关性：尽管了解OA发病机理的最新进展，但对最普遍的联合疾病的疾病改良疗法尚无。检查miR-140在MSC和软骨细胞中的作用可能会提供有关软骨细胞生物学的新见解，也可以为骨关节炎的组织工程和治疗策略的新方法提供基础。