MSC-derived exosomes promote proliferation and inhibit apoptosis of chondrocytes via lncRNA-KLF3-AS1/miR-206/GIT1 axis in osteoarthritis

Abstract

Background: Exosomes secreted by human mesenchymal stem cells (hMSCs) have been shown to promote cartilage regeneration. This study aimed to explore whether exosomal lncRNA-KLF3-AS1 derived from hMSCs can promote chondrocyte proliferation via miR-206/GIT1 axis in osteoarthritis (OA). Methods: hMSCs and MSC-derived exosomes (MSC-exo) were prepared for morphological observation and identification by transmission electron microscopy (TEM) and flow cytometry. IL-1β-induced OA chondrocytes and collagenase-induced mouse OA model were established for the further experiments. Luciferase activity assay was performed to test whether miR-206 could bind to KLF3-AS1 or GIT1. Cell proliferation and apoptosis were evaluated by CCK-8 assay and flow cytometry, respectively. Results: MSC-Exos increased chondrogenic genes Col2a1 (type II collagen alpha 1) and aggrecan, decreased hondrocyte hypertrophy markers MMP-13 (matrix metalloproteinase-13) and Runx2 (runtrelated transcription factor 2) in chondrocytes isolated from OA model mice. Furthermore, MSC-Exos attenuated IL-1β-induced chondrocyte proliferation inhibition and apoptosis induction. Moreover, MSC KLF3-AS1 -Exos (exosomes derived from KLF3-AS1-overexpressing-MSCs) ameliorated IL-1β-induced chondrocyte injury. Results also demonstrated that KLF3-AS1 acted as a competitive endogenous RNA (ceRNA) by sponging miR-206 to facilitate GIT1 expression. In addition, miR-206 overexpression and GIT1 knockdown reversed MSC KLF3-AS1 -Exos-mediated attenuation of chondrocyte injury. Conclusion: Exosomal KLF3-AS1 derived from MSCs involved in MSC-Exos-mediated chondrocyte proliferation induction and chondrocyte apoptosis inhibition via miR-206/GIT1 axis. Abbreviation: G-protein-coupled receptor kinase interacting protein-1 (GIT1) ARTICLE HISTORY

Exosomes in osteoarthritis and cartilage injury: advanced development and potential therapeutic strategies

“…There are two ways to load drugs into exosomes, one is to load drugs into the donor cell of exosomes, such as using transfection and co-incubation; the other is to load drugs into exosomes after they are secreted, such as direct mixing, which the loading efficacy is a big concern. At present, most researchers prefer to obtain the exosomes for OA therapy with high expression of miRNA or lncRNA from modified MSCs [22,[44][45][46] . Exosomal miR-92a-3p [22] , exosomal lncRNA-KLF3-AS1 [44] , exosomal miR-140-5p [45] and exosomal miR-320c [46] from transfected MSCs have been reported to have significant therapeutic effects on OA in vivo and in vitro.…”

Exosomal long noncoding RNAs in aging and age‐related diseases

“…MSCs have been found to play a role in articular cartilage repair in OA . Liu et al found that MSC‐Exos increased the expression of the chondrogenic genes Col2α1 (Type II collagen alpha 1) and aggrecan in chondrocytes of OA model mice decreased the markers of cartilage hypertrophy matrix metalloproteinase (MMP)‐13 and runt‐related transcription factor 2, and also reduced the IL‐1β‐induced apoptosis and inhibition of chondrocyte proliferation. Further studies showed that lncRNA KLF3 Antisense RNA 1 (KLF3‐AS1) in MSC‐Exos promoted the G‐protein‐coupled receptor kinase interacting protein‐1 (GIT1) expression through a miRNA‐206 sponge effect and alleviated the IL‐1β‐induced chondrocyte apoptosis and inhibition of proliferation.…”

Mesenchymal stem cell derived exosomes enhance lymphangiogenesis via exosomal transfer of Ang-2/Tie2

“…MSC-Ex exert effects via delivering MSC-derived cargos (i.e., DNAs, mRNAs, miRNAs, and proteins) to host cells, which lead to changed behaviors of recipient cells to enhance tissue repair. (Liu, Lin et al, 2018, Mayourian, Ceholski et al, 2018, Valadi, Ekstrom et al, 2007 Thus, it will be necessary to determine the factors that control hucMSC-Ex mediated lymphangiogenesis and their antilymphedemic effects. So far antilymphedema studies have only used bone marrow MSC (BM-MSCs) and ADSCs.…”