Regulation of microRNA-221, -222, -21 and -27 in articular cartilage subjected to abnormal compressive forces.

The Journal of Physiology Pub Date : 2021-01-01 Epub Date: 2020-10-31 DOI:10.1113/JP279810
Paulina S Stadnik, Sophie J Gilbert, Jessica Tarn, Sarah Charlton, Andrew J Skelton, Matthew J Barter, Victor C Duance, David A Young, Emma J Blain
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引用次数: 0

Abstract

Key points: microRNAs (miRs) are small non-coding molecules that regulate post-transcriptional target gene expression. miRs are involved in regulating cellular activities in response to mechanical loading in all physiological systems, although it is largely unknown whether this response differs with increasing magnitudes of load. miR-221, miR-222, miR-21-5p and miR-27a-5p were significantly increased in ex vivo cartilage explants subjected to increasing load magnitude and in in vivo joint cartilage exposed to abnormal loading. TIMP3 and CPEB3 are putative miR targets in chondrocytes Identification of mechanically regulated miRs that have potential to impact on tissue homeostasis provides a mechanism by which load-induced tissue behaviour is regulated, in both health and pathology, in all physiological systems.

Abstract: MicroRNAs (miRs) are small non-coding molecules that regulate post-transcriptional target gene expression and are involved in mechano-regulation of cellular activities in all physiological systems. It is unknown whether such epigenetic mechanisms are regulated in response to increasing magnitudes of load. The present study investigated mechano-regulation of miRs in articular cartilage subjected to 'physiological' and 'non-physiological' compressive loads in vitro as a model system and validated findings in an in vivo model of abnormal joint loading. Bovine full-depth articular cartilage explants were loaded to 2.5 MPa (physiological) or 7 MPa (non-physiological) (1 Hz, 15 min) and mechanically-regulated miRs identified using next generation sequencing and verified using a quantitative PCR. Downstream targets were verified using miR-specific mimics or inhibitors in conjunction with 3'-UTR luciferase activity assays. A subset of miRs were mechanically-regulated in ex vivo cartilage explants and in vivo joint cartilage. miR-221, miR-222, miR-21-5p and miR-27a-5p were increased and miR-483 levels decreased with increasing load magnitude. Tissue inhibitor of metalloproteinase 3 (TIMP3) and cytoplasmic polyadenylation element binding protein 3 (CPEB3) were identified as putative downstream targets. Our data confirm miR-221 and -222 mechano-regulation and demonstrates novel mechano-regulation of miR-21-5p and miR-27a-5p in ex vivo and in vivo cartilage loading models. TIMP3 and CPEB3 are putative miR targets in chondrocytes. Identification of specific miRs that are regulated by increasing load magnitude, as well as their potential to impact on tissue homeostasis, has direct relevance to other mechano-sensitive physiological systems and provides a mechanism by which load-induced tissue behaviour is regulated, in both health and pathology.

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microRNA-221, -222, -21和-27在关节软骨承受异常压缩力时的调控作用。
重点:microRNAs (miRs)是调节转录后靶基因表达的非编码小分子。在所有的生理系统中,miRs参与调节细胞对机械负荷的反应,尽管这种反应是否随着负荷的增加而变化在很大程度上是未知的。miR-221、miR-222、miR-21-5p和miR-27a-5p在体外软骨外植体和体内关节软骨暴露于异常负荷时显著升高。TIMP3和CPEB3是软骨细胞中推测的miR靶点。机械调节的miRs对组织稳态有潜在的影响,这为在所有生理系统中调节负荷诱导的组织行为提供了一种机制。摘要:MicroRNAs (miRs)是一种调节转录后靶基因表达的非编码小分子,参与所有生理系统中细胞活动的机械调节。目前尚不清楚这种表观遗传机制是否会随着负荷的增加而受到调节。本研究在体外作为模型系统研究了受“生理”和“非生理”压缩载荷的关节软骨中miRs的机械调节,并在体内异常关节载荷模型中验证了研究结果。将牛全深度关节软骨外植体加载到2.5 MPa(生理)或7 MPa(非生理)(1 Hz, 15 min),并通过下一代测序和定量PCR鉴定机械调节的miRs。下游靶标使用mir特异性模拟物或抑制剂结合3'-UTR荧光素酶活性测定进行验证。一部分miRs在体外软骨外植体和体内关节软骨中受到机械调节。随着负荷强度的增加,miR-221、miR-222、miR-21-5p和miR-27a-5p水平升高,miR-483水平降低。组织金属蛋白酶3抑制剂(TIMP3)和细胞质多腺苷化元件结合蛋白3 (CPEB3)被确定为可能的下游靶点。我们的数据证实了miR-221和-222的机械调节,并在离体和体内软骨负荷模型中展示了miR-21-5p和miR-27a-5p的新型机械调节。TIMP3和CPEB3是软骨细胞中推测的miR靶点。鉴定受增加负荷大小调节的特定miRs,以及它们对组织稳态的潜在影响,与其他机械敏感的生理系统直接相关,并提供了一种在健康和病理方面调节负荷诱导的组织行为的机制。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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