miR-3960 from Mesenchymal Stem Cell-Derived Extracellular Vesicles Inactivates SDC1/Wnt/β-Catenin Axis to Relieve Chondrocyte Injury in Osteoarthritis by Targeting PHLDA2

Abstract

Osteoarthritis (OA) is a serious disease of the articular cartilage characterized by excessive inflammation. Lately, mesenchymal stem cell- (MSC-) derived extracellular vesicles (EVs) have been proposed as a novel strategy for the treatment of OA. We aimed to investigate the effects of EV-encapsulated miR-3960 derived from MSCs on chondrocyte injury in OA. The cartilage tissues from OA patients were collected to experimentally determine expression patterns of miR-3960, PHLDA2, SDC1, and β-catenin. Next, luciferase assay was implemented to testify the binding affinity among miR-3960 and PHLDA2. EVs were isolated from MSCs and cocultured with IL-1β-induced OA chondrocytes. Afterwards, cellular biological behaviors and levels of extracellular matrix- (ECM-) related protein anabolic markers (collagen II and aggrecan), catabolic markers (MMP13 and ADAMTS5), and inflammatory factors (IL-6 and TNF-α) in chondrocytes were assayed upon miR-3960 and/or PHLDA2 gain- or loss-of-function. Finally, the effects of miR-3960 contained in MSC-derived EVs in OA mouse models were also explored. MSCs-EVs could reduce IL-1β-induced inflammatory response and extracellular matrix (ECM) degradation in chondrocytes. miR-3960 expression was downregulated in cartilage tissues of OA patients but enriched in MSC-derived EVs. miR-3960 could target and inhibit PHLDA2, which was positively correlated with SDC1 and Wnt/β-catenin pathway activation. miR-3960 shuttled by MSC-derived EVs protected against apoptosis and ECM degradation in chondrocytes. In vivo experiment also confirmed that miR-3960 alleviated chondrocyte injury in OA. Collectively, MSC-derived EV-loaded miR-3960 downregulated PHLDA2 to inhibit chondrocyte injury via SDC1/Wnt/β-catenin.

Strategies in product engineering of mesenchymal stem cell-derived exosomes: unveiling the mechanisms underpinning the promotive effects of mesenchymal stem cell-derived exosomes

“…However, miR-3960, enriched in the exosomes secreted by MSCs, can be delivered to chondrocyte cells, subsequently degrading PHLDA2 mRNA. Moreover, the expression profiles of miR-3960, PHLDA2, SDC1, and β-catenin in cartilage tissues from OA patients align with findings from their prospective study ( Ye et al, 2022 ).Xia Qingqing et al identified a negative correlation between miR-125a-5p and E2F2 in the cartilage of traumatic osteoarthritis patients and in a corresponding mouse model. The exosomes derived from bone marrow MSCs, which are taken up by chondrocytes in mice, contain miR-125a-5p that suppresses E2F2 expression.…”

Analysis of extracellular vesicle microRNA profiles reveals distinct blood and lymphatic endothelial cell origins

“…The qPCR validation of three selected miRNAs with a high logFC (smaller than–6) between Cell and EV for ECs and a selection of control cells indicated that the identified subset of miRNAs may be secreted actively by other cell types as well hinting towards EV specific miRNA species. All three miRNAs were reported to be associated to EVs not only from EC but for example in plasma derived EVs or in cancer related studies (Umair et al., 2022 ; Ye et al., 2022 ; Zhong et al., 2021 ). MicroRNA miR‐3960 was shown to promote the differentiation process of vascular smooth muscle cells into osteoblasts (Xia et al., 2015 ).…”

Mesenchymal stem cell-derived exosomes as a promising cell-free therapy for knee osteoarthritis

“…It was reported that BMMSC-derived exosomal miR-320c increased chondrocyte proliferation by increasing the expression of SOX9 and decreasing MMP-13 levels ( Sun et al, 2019 ). Another study showed that BMMSC-Exos could upregulate the levels of COL2 and aggrecan alongside downregulate ADAMTS-5 and MMP-13 expression by encapsulating miR-3960 ( Ye et al, 2022 ). Moreover, BMMSC-derived exosomal miR-125a-5p was demonstrated to alleviate chondrocyte ECM degradation via inhibiting E2F2 in post-traumatic OA ( Xia et al, 2021 ).…”