Krembil Research Institute, University Health Network, and, University of Toronto, Toronto, Ontario, Canada.
Krembil Research Institute, University Health Network, Toronto, Ontario, Canada.
Arthritis Rheumatol. 2021 Mar;73(3):426-439. doi: 10.1002/art.41552. Epub 2021 Feb 8.
MicroRNA-34a-5p (miR-34a-5p) expression is elevated in the synovial fluid of patients with late-stage knee osteoarthritis (OA); however, its exact role and therapeutic potential in OA remain to be fully elucidated. This study was undertaken to examine the role of miR-34a-5p in OA pathogenesis.
Expression of miR-34a-5p was determined in joint tissues and human plasma (n = 71). Experiments using miR-34a-5p mimic or antisense oligonucleotide (ASO) treatment were performed in human OA chondrocytes, fibroblast-like synoviocytes (FLS) (n = 7-9), and mouse OA models, including destabilization of the medial meniscus (DMM; n = 22) and the accelerated, more severe model of mice fed a high-fat diet and subjected to DMM (n = 11). Wild-type (WT) mice (n = 9) and miR-34a-knockout (KO) mice (n = 11) were subjected to DMM. Results were expressed as the mean ± SEM and analyzed by t-test or analysis of variance, with appropriate post hoc tests. P values less than 0.05 were considered significant. RNA sequencing was performed on WT and KO mouse chondrocytes.
Expression of miR-34a-5p was significantly increased in the plasma, cartilage, and synovium of patients with late-stage OA and in the cartilage and synovium of mice subjected to DMM. Plasma miR-34a-5p expression was significantly increased in obese patients with late-stage OA, and in the plasma and knee joints of mice fed a high-fat diet. In human OA chondrocytes and FLS, miR-34a-5p mimic increased key OA pathology markers, while miR-34a-5p ASO improved cellular gene expression. Intraarticular miR-34a-5p mimic injection induced an OA-like phenotype. Conversely, miR-34a-5p ASO injection imparted cartilage-protective effects in the DMM and high-fat diet/DMM models. The miR-34a-KO mice exhibited protection against DMM-induced cartilage damage. RNA sequencing of WT and KO chondrocytes revealed a putative miR-34a-5p signaling network.
Our findings provide comprehensive evidence of the role and therapeutic potential of miR-34a-5p in OA.
在晚期膝骨关节炎（OA）患者的滑液中，miR-34a-5p 的表达升高；然而，其在 OA 中的确切作用和治疗潜力仍有待充分阐明。本研究旨在探讨 miR-34a-5p 在 OA 发病机制中的作用。
在关节组织和人血浆中（n=71）测定 miR-34a-5p 的表达。在人 OA 软骨细胞、成纤维样滑膜细胞（FLS）（n=7-9）以及包括内侧半月板不稳定（DMM；n=22）和高脂肪饮食喂养并接受 DMM 的加速、更严重的小鼠模型中进行 miR-34a-5p 模拟物或反义寡核苷酸（ASO）治疗实验。对野生型（WT）小鼠（n=9）和 miR-34a 敲除（KO）小鼠（n=11）进行 DMM。结果表示为均值±SEM，并通过 t 检验或方差分析进行分析，并进行适当的事后检验。P 值小于 0.05 被认为具有统计学意义。对 WT 和 KO 小鼠软骨细胞进行 RNA 测序。
晚期 OA 患者的血浆、软骨和滑膜以及 DMM 小鼠的软骨和滑膜中 miR-34a-5p 的表达显著增加。肥胖晚期 OA 患者的血浆 miR-34a-5p 表达以及高脂肪饮食喂养的小鼠的血浆和膝关节中，miR-34a-5p 的表达均显著增加。在人 OA 软骨细胞和成纤维样滑膜细胞中，miR-34a-5p 模拟物增加了关键的 OA 病理标志物，而 miR-34a-5p ASO 改善了细胞基因表达。关节内 miR-34a-5p 模拟物注射诱导 OA 样表型。相反，miR-34a-5p ASO 注射在 DMM 和高脂肪饮食/DMM 模型中具有软骨保护作用。miR-34a-KO 小鼠对 DMM 诱导的软骨损伤具有保护作用。WT 和 KO 软骨细胞的 RNA 测序揭示了一个潜在的 miR-34a-5p 信号网络。
我们的研究结果为 miR-34a-5p 在 OA 中的作用和治疗潜力提供了全面的证据。
