Exosomes from osteoarthritic fibroblast-like synoviocytes promote cartilage ferroptosis and damage via delivering microRNA-19b-3p to target SLC7A11 in osteoarthritis

Abstract

ObjectiveOur previous studies revealed that normal synovial exosomes promoted chondrogenesis, and microRNA (miR)-19b-3p independently related to osteoarthritis (OA) risk. Subsequently, this study intended to further explore the effect of OA fibroblast-like synoviocyte (OA-FLS) exosomal miR-19b-3p on OA ferroptosis and its potential mechanisms.MethodsInterleukin (IL)-1β-stimulated chondrocytes and medial meniscus surgery were used to construct the OA cellular model and the OA rat model, respectively. OA-FLS exosomes with/without miR-19b-3p modification were added to the IL-1β-stimulated chondrocytes and OA rat models, followed by direct miR-19b-3p mimic/inhibitor transfection with/without SLC7A11 overexpression plasmids. miR-19b-3p, ferroptosis-related markers (malondialdehyde (MDA), glutathione (GSH)/oxidized glutathione (GSSG), ferrous ion (Fe2+), glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), and acyl-CoA synthetase long-chain family member 4 (ACSL4)), mitochondrial membrane potential (MMP), and reactive oxygen species (ROS) levels were detected.ResultsEnhanced ferroptosis reflected by dysregulated ferroptosis-related markers, a reduced MMP, and an increased ROS was observed in cartilage tissues from OA patients vs. controls, IL-1β-stimulated chondrocytes vs. normal ones, and OA rat models vs. sham, so did miR-19b-3p. OA-FLS exosomes promoted MDA, Fe2+, ACSL4, and ROS but reduced cell viability, GSH/GSSG, GPX4, SLC7A11, and MMP in IL-1β-stimulated chondrocytes, whose effect was enhanced by miR-19b-3p mimics and attenuated by miR-19b-3p inhibitors. miR-19b-3p negatively regulated SLC7A11 and directly bound to SLC7A11 via luciferase reporter gene assay. Furthermore, SLC7A11 overexpression weakened miR-19b-3p mimics’ effect on ferroptosis-related markers, MMP, or ROS in IL-1β-stimulated chondrocytes. OA-FLS exosomes also induced cartilage damage and ferroptosis in OA rats whose influence was tempered by miR-19b-3p inhibitors.ConclusionOA-FLS exosomal miR-19b-3p enhances cartilage ferroptosis and damage by sponging SLC7A11 in OA, indicating a potential linkage among synovium, cartilage, and ferroptosis during the OA process.

New insight of the pathogenesis in osteoarthritis: the intricate interplay of ferroptosis and autophagy mediated by mitophagy/chaperone-mediated autophagy

“…This study also found that compared with the normal control group, the expression of ferroptosis related markers SLC7A11, GSH, and GPX4 in cartilage tissue of OA patients decreased, while the expression of MDA, Fe 2+ , ACSL4, and miR-19b-3p in extracellular vesicles increased (Kong et al, 2023). Simultaneously using IL-1β Inducing chondrocytes to construct an OA chondrocyte degeneration model, the results showed a significant decrease in chondrocyte viability, with reduced expression levels of SCL7A11, GSH/GSSG, GPX4, and MMP, while increased expression levels of MDA, Fe 2+ , ASCL4, and ROS (Kong et al, 2023). The OA-FLS extracellular vesicles can further reduce the chondrocyte viability of the OA chondrocyte degeneration model, increase the downregulated expression of SCL7A11, GSH/GSSG, GPX4, and MMP in the OA chondrocyte degeneration model, and increase the upregulated expression of MDA, Fe 2+ , ASCL4, and ROS in the OA chondrocyte degeneration model (Kong et al, 2023).…”

“…Simultaneously using IL-1β Inducing chondrocytes to construct an OA chondrocyte degeneration model, the results showed a significant decrease in chondrocyte viability, with reduced expression levels of SCL7A11, GSH/GSSG, GPX4, and MMP, while increased expression levels of MDA, Fe 2+ , ASCL4, and ROS (Kong et al, 2023). The OA-FLS extracellular vesicles can further reduce the chondrocyte viability of the OA chondrocyte degeneration model, increase the downregulated expression of SCL7A11, GSH/GSSG, GPX4, and MMP in the OA chondrocyte degeneration model, and increase the upregulated expression of MDA, Fe 2+ , ASCL4, and ROS in the OA chondrocyte degeneration model (Kong et al, 2023). The intervention of ferroptosis inhibitor Fer-1 weakened the influence of OA-FLS extracellular vesicles on the above indicators.…”

“…The expression level of GPX4 may have a potential connection with the progression of OA (Wang et al, 2022). Kong et al (Kong et al, 2023) found in their study on the effect and potential mechanism of the extracellular vesicle miR-19b-3p of OA fibroblasts like synovial cells (OA FLS) on OA ferroptosis that the extracellular vesicle miR-19b-3p of OA FLS promotes cartilage ferroptosis and tissue damage by sponging SLC7A11 in OA (Kong et al, 2023). This study also found that compared with the normal control group, the expression of ferroptosis related markers SLC7A11, GSH, and GPX4 in cartilage tissue of OA patients decreased, while the expression of MDA, Fe 2+ , ACSL4, and miR-19b-3p in extracellular vesicles increased (Kong et al, 2023).…”

“…Kong et al (Kong et al, 2023) found in their study on the effect and potential mechanism of the extracellular vesicle miR-19b-3p of OA fibroblasts like synovial cells (OA FLS) on OA ferroptosis that the extracellular vesicle miR-19b-3p of OA FLS promotes cartilage ferroptosis and tissue damage by sponging SLC7A11 in OA (Kong et al, 2023). This study also found that compared with the normal control group, the expression of ferroptosis related markers SLC7A11, GSH, and GPX4 in cartilage tissue of OA patients decreased, while the expression of MDA, Fe 2+ , ACSL4, and miR-19b-3p in extracellular vesicles increased (Kong et al, 2023). Simultaneously using IL-1β Inducing chondrocytes to construct an OA chondrocyte degeneration model, the results showed a significant decrease in chondrocyte viability, with reduced expression levels of SCL7A11, GSH/GSSG, GPX4, and MMP, while increased expression levels of MDA, Fe 2+ , ASCL4, and ROS (Kong et al, 2023).…”

“…Indicating that OA-FLS extracellular vesicles promote the ferroptosis in IL-1β induced OA chondrocyte model. The author further used miR-19b-3p mimetics to overexpress miR-19b-3p, and the results showed that overexpression of miR-19b-3p reduced the chondrocyte viability of the OA chondrocyte degeneration model and reduced the expression of GPX4, GSH/GSSG, SLC7A11, and MMP in the OA chondrocyte degeneration model, but increased the expression of ACSL4, MDA, Fe 2+ , and ROS in the OA chondrocyte degeneration model (Kong et al, 2023). Inhibiting the expression of miR-19b-3p with miR-19b-3p inhibitors showed the opposite result, confirming that the miR-19b-3p in extracellular vesicles of OA FLS may be a key target leading to ferroptosis in OA.…”