机械敏感的 microRNAs 受人类软骨的合成代谢和分解代谢加载调节。

Mechanosensitive MiRs regulated by anabolic and catabolic loading of human cartilage.

机构信息

Research Centre for Experimental Orthopaedics, Orthopaedic University Hospital Heidelberg, Heidelberg, Germany.

Department of Orthopaedics and Rehabilitation, Oregon Health & Science University, Portland, OR, United States.

出版信息

Osteoarthritis Cartilage. 2019 Aug;27(8):1208-1218. doi: 10.1016/j.joca.2019.04.010. Epub 2019 Apr 19.

DOI:10.1016/j.joca.2019.04.010

PMID:31009748

Abstract

OBJECTIVE

Elucidation of whether miRs are involved in mechanotransduction pathways by which cartilage is maintained or disturbed has a particular importance in our understanding of osteoarthritis (OA) pathophysiology. The aim was to investigate whether mechanical loading influences global miR-expression in human chondrocytes and to identify mechanosensitive miRs responding to beneficial and non-beneficial loading regimes as potential to obtain valuable diagnostic or therapeutic targets to advance OA-treatment.

METHOD

Mature tissue-engineered human cartilage was subjected to two distinct loading regimes either stimulating or suppressing proteoglycan-synthesis, before global miR microarray analysis. Promising candidate miRs were selected, re-evaluated by qRT-PCR and tested for expression in human healthy vs OA cartilage samples.

RESULTS

After anabolic loading, miR microarray profiling revealed minor changes in miR-expression while catabolic stimulation produced a significant regulation of 80 miRs with a clear separation of control and compressed samples by hierarchical clustering. Cross-testing of selected miRs revealed that miR-221, miR-6872-3p, miR-6723-5p were upregulated by both loading conditions while others (miR-199b-5p, miR-1229-5p, miR-1275, miR-4459, miR-6891-5p, miR-7150) responded specifically after catabolic loading. Mechanosensitivity of miR-221 correlated with pERK1/2-activation induced by both loading conditions. The miR-response to loading was transient and a constitutive deregulation of mechano-miRs in OA vs healthy articular cartilage was not observed.

CONCLUSIONS

MiRs with broader vs narrower mechanosensitivity were discovered and the first group of mechanosensitive miRs characteristic for non-beneficial loading was defined that may shape the proteome differentially when cartilage tissue is disturbed. The findings prompt future investigations into miR-relevance for mechano-responsive pathways and the corresponding miR-target molecules.

摘要

目的

阐明 miR 是否参与软骨维持或破坏的力学转导途径，这对于我们理解骨关节炎（OA）病理生理学具有特殊意义。本研究旨在探讨机械负荷是否影响人软骨细胞的整体 miR 表达，并确定对有益和无益负荷反应的机械敏感 miR，以作为获得有价值的诊断或治疗靶点，推进 OA 治疗的可能性。

方法

成熟的组织工程化人软骨在受到两种不同的负荷刺激，分别刺激或抑制蛋白聚糖合成后，进行整体 miR 微阵列分析。选择有前途的候选 miR，通过 qRT-PCR 重新评估，并在健康对照和 OA 软骨样本中测试其表达。

结果

在合成代谢负荷后，miR 微阵列分析显示 miR 表达的微小变化，而分解代谢刺激则导致 80 个 miR 的显著调节，通过层次聚类清楚地分离了对照和压缩样本。对选定 miR 的交叉测试表明，miR-221、miR-6872-3p、miR-6723-5p 被两种负荷条件上调，而其他 miR（miR-199b-5p、miR-1229-5p、miR-1275、miR-4459、miR-6891-5p、miR-7150）则仅在分解代谢负荷后特异性应答。miR-221 的力学敏感性与两种负荷条件诱导的 pERK1/2 激活相关。miR 对负荷的反应是短暂的，OA 与健康关节软骨之间并未观察到机械 miR 的持续失调。