miR-92a-3p-inspired shRNA exhibits pro-chondrogenic and chondrocyte protective effects in osteoarthritis treatment through targeting SMAD6/7.

IF 2.4 3区医学 Q3 ENDOCRINOLOGY & METABOLISM Journal of Bone and Mineral Metabolism Pub Date : 2024-01-01 Epub Date: 2023-12-06 DOI:10.1007/s00774-023-01474-3

Chenhuang Zheng, Kazuto Hoshi, Atsuhiko Hikita

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Abstract

Introduction: Osteoarthritis (OA) compromises patients' quality of life and requires further study. Although miR-92a-3p was reported to possess chondroprotective effects, the underlying mechanism requires further clarification. The objectives of this study were to elucidate the mechanism by which miR-92a-3p alleviates OA and to examine the efficacy of shRNA-92a-3p, which was designed based on mature miR-92a-3p.

Materials and methods: TargetScan and luciferase reporter assay were used to predict the target of miR-92a-3p. Adipose-derived stem cells (ADSCs) were transfected with miR-92a-3p/miR-NC mimic for the analysis of chondrogenic biomarkers and SMAD proteins. ADSCs and osteoarthritic chondrocytes were transduced with shRNA-92a-3p for the analysis of chondrogenic biomarkers and SMAD proteins. OA was surgically induced in C57BL/6JJcl mice, and ADSCs with/without shRNA-92a-3p transduction were intra-articularly injected for the assessment of cartilage damage.

Results: SMAD6 and SMAD7 were predicted as direct targets of miR-92a-3p by TargetScan and luciferase reporter assay. Transfection of the miR-92a-3p mimic resulted in a decrease in SMAD6 and SMAD7 levels and an increase in phospho-SMAD2/3, phospho-SMAD1/5/9, SOX9, collagen type II, and aggrecan levels in ADSCs. Furthermore, shRNA-92a-3p decreased SMAD6 and SMAD7 levels, and increased phospho-SMAD2/3, phospho-SMAD1/5/9, SOX9, collagen type II, and aggrecan levels in ADSCs and osteoarthritic chondrocytes. Additionally, ADSC-shRNA-92a-3p-EVs reduced the rate of decrease of SOX9, collagen type II, and aggrecan in osteoarthritic chondrocytes. In mice with surgically induced OA, shRNA-92a-3p-treated ADSCs alleviated cartilage damage more effectively than nontreated ADSCs.

Conclusions: miR-92a-3p and shRNA-92a-3p exhibit therapeutic effects in treating OA by targeting SMAD6 and SMAD7, thereby enhancing TGF-β signaling.

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miR-92a-3p-inspired shRNA 通过靶向 SMAD6/7 在骨关节炎治疗中表现出促进软骨生成和保护软骨细胞的作用。

导言：骨关节炎（OA）影响患者的生活质量，需要进一步研究。尽管有报道称 miR-92a-3p 具有保护软骨的作用，但其基本机制仍需进一步阐明。本研究的目的是阐明 miR-92a-3p 缓解 OA 的机制，并研究基于成熟 miR-92a-3p 设计的 shRNA-92a-3p 的疗效：TargetScan 和荧光素酶报告实验用于预测 miR-92a-3p 的靶点。用 miR-92a-3p/miR-NC mimic 转染脂肪源性干细胞（ADSCs），分析软骨生成生物标志物和 SMAD 蛋白。用 shRNA-92a-3p 转染 ADSCs 和骨关节炎软骨细胞，分析软骨生成生物标志物和 SMAD 蛋白。通过手术诱导C57BL/6JJcl小鼠患上OA，将转导/未转导shRNA-92a-3p的ADSCs关节内注射，以评估软骨损伤情况：结果：通过 TargetScan 和荧光素酶报告实验预测 SMAD6 和 SMAD7 是 miR-92a-3p 的直接靶点。转染 miR-92a-3p mimic 后，ADSCs 中的 SMAD6 和 SMAD7 水平下降，磷酸-SMAD2/3、磷酸-SMAD1/5/9、SOX9、II 型胶原和 aggrecan 水平上升。此外，在 ADSCs 和骨关节炎软骨细胞中，shRNA-92a-3p 降低了 SMAD6 和 SMAD7 水平，增加了磷酸化-SMAD2/3、磷酸化-SMAD1/5/9、SOX9、胶原蛋白 II 型和 aggrecan 水平。此外，ADSC-shRNA-92a-3p-EVs 还能降低骨关节炎软骨细胞中 SOX9、II 型胶原和 aggrecan 的下降速度。结论：miR-92a-3p和shRNA-92a-3p通过靶向SMAD6和SMAD7，从而增强TGF-β信号传导，对治疗OA具有治疗作用。

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来源期刊

Journal of Bone and Mineral Metabolism 医学-内分泌学与代谢

CiteScore

6.30

自引率

3.00%

发文量

审稿时长

6-12 weeks

期刊介绍： The Journal of Bone and Mineral Metabolism (JBMM) provides an international forum for researchers and clinicians to present and discuss topics relevant to bone, teeth, and mineral metabolism, as well as joint and musculoskeletal disorders. The journal welcomes the submission of manuscripts from any country. Membership in the society is not a prerequisite for submission. Acceptance is based on the originality, significance, and validity of the material presented. The journal is aimed at researchers and clinicians dedicated to improvements in research, development, and patient-care in the fields of bone and mineral metabolism.