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

基本信息

批准号：
10115614
负责人：
HIROSHI ASAHARA
金额：
$ 41.08万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-07-01 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10115614
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

项目摘要

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.

抽象的软骨细胞在肢体发育，产后骨骼生长和维持中起重要作用成人生活期间软骨组织。在软骨细胞中基因表达的失调是关键事件骨关节炎的发病机理（OA）。 microRNA是一个非编码RNA的家族，在进化上是通过转录后机制保守和调节基因表达。许多mir显示特异性组织表达模式，表明这些miR在组织特异性生理学或发展过程。但是，软骨细胞特异性miR，它们的上游分子信号和靶标基因才开始被鉴定。我们和其他人报告说，Mir-140是软骨特异性miRNA，在软骨发育和稳态中起着至关重要的作用，而OA的表达降低有助于发病机理。在我们的初步实验中，miR-455（miR-455-5p和miR-455-3p）的链以及miR-140 被确定为Sox9的靶标，Sox9是软骨特异性基因的主要转录因子。 mir的表达与正常软骨相比，在人OA软骨中下调了455-5p/3p。使用CRISPR/CAS9 系统，我们生成了miR-455-5p/3p敲除小鼠。这些小鼠在6个月时显示出严重的OA表型。微阵列分析揭示了软骨细胞中的一组可能的miR-455靶基因，包括HIF2A，其中，这些基因是OA发病机理中的关键转录因子，也是MiR455链的靶标。重要的是，HIF2A 在6个月时，在miR-455无效小鼠关节软骨中，几乎所有软骨细胞中的几乎所有软骨细胞都大大增加。这些观察结果支持mir-455-5p/3p是软骨的关键调节剂的假设体内平衡及其表达和功能的变化在影响的疾病中起着重要作用关节软骨。我们提出以下具体目标： AIM 1：使用miR-455 KO小鼠和人类分析miR-455-5p/3p在OA发病机理中的功能关节软骨细胞。目标2：在关节软骨细胞中识别调节软骨的下游目标直接miR-455-5p/3p 稳态和OA发病机理。 AIM 3：检查关节内miR-455S在OA动物模型中给药的治疗作用。拟议的研究有可能揭示控制的重要新调节途径软骨发展和稳态，并开放有关疾病机制和治疗性的新见解干预措施。

项目成果

期刊论文数量（46）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Transcription activator-like effector nuclease-mediated transduction of exogenous gene into IL2RG locus.

DOI：
10.1038/srep05043
发表时间：
2014-05-23
期刊：
Scientific reports
影响因子：
4.6
作者：
Matsubara Y;Chiba T;Kashimada K;Morio T;Takada S;Mizutani S;Asahara H
通讯作者：
Asahara H

Expression of MicroRNA-146a in osteoarthritis cartilage.

DOI：
10.1002/art.24404
发表时间：
2009-04
期刊：
ARTHRITIS AND RHEUMATISM
影响因子：
0
作者：
Yamasaki, Keiichiro;Nakasa, Tomoyuki;Miyaki, Shigeru;Ishikawa, Masakazu;Deie, Masataka;Adachi, Nobuo;Yasunaga, Yuji;Asahara, Hiroshi;Ochi, Mitsuo
通讯作者：
Ochi, Mitsuo

DNA binding-dependent glucocorticoid receptor activity promotes adipogenesis via Krüppel-like factor 15 gene expression.