The role of microRNAs in cartilage homeostasis and osteoarthritis arthritis treatment

microRNA在软骨稳态和骨关节炎治疗中的作用

• 批准号: 9906170
• 负责人: HIROSHI ASAHARA
• 金额: $ 42.35万
• 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 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9906170
• 关键词: Adult; Affect; Animal Model; Arthritis; Bioinformatics; CRISPR/Cas technology; Cartilage; Cells; Chondrocytes; Chondrogenesis; Degenerative polyarthritis; Developmental Process; Disease; Disease Progression; Event; Experimental Models; Extracellular Matrix; Family; Gene Expression; Genes; Growth; High Mobility Group Proteins; Histologic; Homeostasis; Human; In Vitro; Inflammatory; Interleukin-1 beta; Knee joint; Knockout Mice; Life; Limb Development; Maintenance; Mesenchymal; MicroRNAs; Microarray Analysis; Molecular; Movement; Mus; Neonatal; Nucleotides; Operative Surgical Procedures; Pathogenesis; Pattern; Pharmaceutical Preparations; Phenotype; Physiologic Ossification; Physiological Processes; Play; Process; Regulation; Regulator Genes; Regulatory Pathway; Reporting; Role; Signal Transduction; Surgical Models; Surveys; Synovial joint; System; Testing; Therapeutic Effect; Therapeutic Intervention; Tissues; Untranslated RNA; Work; arthropathies; articular cartilage; bone; cartilage development; early onset; experimental study; insight; novel therapeutic intervention; overexpression; postnatal; prevent; programs; skeletal; transcription factor

ABSTRACT Chondrocytes play an important role in limb development, postnatal skeletal growth and in maintaining cartilage tissues during adult life. Dysregulation of gene expression in chondrocytes is a key event in the pathogenesis of osteoarthritis (OA). MicroRNAs are a family of noncoding RNAs that are evolutionarily conserved and regulate gene expression by posttranscriptional mechanisms. Many miRs show tissue specific expression patterns, suggesting that these miRs play a crucial role in tissue specific physiological or developmental processes. However, chondrocyte specific miRs, their upstream molecular signals, and target genes are only beginning to be identified. We and others reported that miR-140, a cartilage specific miRNA, plays a critical role both in cartilage development and homeostasis while the reduced expression in OA contributes to pathogenesis. In our preliminary experiments, both strands of miR-455 (miR-455-5p and miR-455-3p) as well as miR-140 were identified as targets of Sox9, a main transcription factor for cartilage specific genes. Expression of miR- 455-5p/3p was down-regulated in human OA cartilage compared with normal cartilage. Using CRISPR/Cas9 system, we generated miR-455-5p/3p knockout mice. These mice showed a severe OA phenotype at 6 months. Microarray analyses revealed a set of possible miR-455 target genes in chondrocytes, including HIF2A, which is a key transcription factor in OA pathogenesis, and a target of both strands of miR455. Importantly, HIF2A was strongly increased in almost all chondrocytes in articular cartilage of miR-455 null mice at 6 months. These observations support the hypothesis that miR-455-5p/3p are critical regulators of cartilage homeostasis and that changes in their expression and function play an important role in diseases affecting articular cartilage. We propose the following specific aims: Aim 1: Analyze the function of miR-455-5p/3p in OA pathogenesis using miR-455 KO mice and human articular chondrocytes. Aim 2: Identify in articular chondrocytes the direct miR-455-5p/3p downstream targets that regulate cartilage homeostasis and OA pathogenesis. Aim 3: Examine the therapeutic effects of intraarticular miR-455s administration in animal models of OA. 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.

摘要 软骨细胞在肢体发育、出生后骨骼生长和维持骨骼系统中起着重要作用。 软骨组织在成年期。软骨细胞中基因表达的失调是软骨细胞凋亡的关键事件。 骨关节炎（OA）的发病机制。MicroRNA是一个非编码RNA家族， 通过转录后机制保守和调节基因表达。许多miR显示组织特异性 表达模式，表明这些miR在组织特异性生理或 发展过程。然而，软骨细胞特异性miRs，其上游分子信号和靶点， 基因的识别才刚刚开始。我们和其他人报道了miR-140，一种软骨特异性miRNA， 在软骨发育和体内平衡中起关键作用，而在OA中表达减少， 有助于发病机制。 在我们的初步实验中，miR-455的两条链（miR-455- 5 p和miR-455- 3 p）以及miR-140 被鉴定为Sox 9的靶点，Sox 9是软骨特异性基因的主要转录因子。miR-1的表达 与正常软骨相比，455- 5 p/3 p在人OA软骨中下调。使用CRISPR/Cas9 系统中，我们产生了miR-455- 5 p/3 p敲除小鼠。这些小鼠在6个月时表现出严重的OA表型。 微阵列分析揭示了软骨细胞中一组可能的miR-455靶基因，包括HIF 2A， 是OA发病机制中的关键转录因子，并且是miR 455的两条链的靶标。重要的是，HIF 2A 在6个月时，miR-455缺失小鼠的关节软骨中几乎所有软骨细胞中的TGFmRNA均强烈增加。 这些观察结果支持了miR-455- 5 p/3 p是软骨的关键调节因子的假设。 体内平衡及其表达和功能变化在影响 关节软骨我们提出以下具体目标： 目的1：利用miR-455基因敲除小鼠和人，分析miR-455- 5 p/3 p在OA发病机制中的作用 关节软骨细胞。 目的2：在关节软骨细胞中鉴定调节软骨的直接miR-455- 5 p/3 p下游靶点 稳态和OA发病机制。 目的3：检查关节内施用miR-455 s在OA动物模型中的治疗效果。 拟议的研究有可能揭示重要的新的调控途径， 软骨发育和体内平衡，为疾病机制和治疗提供新的见解 干预措施。