Rohto Advanced Research Hong Kong Sha tin, Hong Kong, Hong Kong
Abstract: Mesenchymal stem cells (MSCs) have low immunogenicity with immunosuppressive properties, making them a promising tool for cell-based therapy. They can exert their anti-inflammatory effects through direct cell-to-cell interaction and secretion of bioactive molecules, including cytokines such as IL-10. The primary function of IL-10 is to suppress inflammation. Thereby, the interaction between MSCs and IL-10 was mutually beneficial and could bring a bigger extent in promoting anti-inflammatory. On the other hand, the use of MSCs extracellular vesicles (EVs) rather than MSCs themselves becomes more attractive approach because there are certain concerns regarding MSCs as a clinical application. In this respect, we aim to enhance the anti-inflammatory properties of MSCs and their EVs by overexpressing IL-10. IL-10 was overexpressed in immortalized MSCs (IL-10-iMSCs) and their EV (IL-10-iMSCs-EV) was isolated. Both IL-10-iMSCs and IL-10-iMSCs-EV were characterized and tested in various in vitro assays for their anti-inflammatory properties. Furthermore, IL-10-iMSCs-EV content was analyzed by RNA sequencing to evaluate potential microRNA for anti-inflammatory role. During cell culture and EV production, inhouse formulated clinical grade serum free MSC medium were used. Our result showed that overexpression of IL-10 was observed in IL-10-iMSCs with downregulation of inflammation-related genes such as IL-6. Through the in vitro assays, IL-10-iMSCs treatment showed a significant reduction in pro-inflammatory genes expression and enhanced anti-inflammatory M2 related markers than normal iMSCs. This promising anti-inflammatory result was further enhanced by IL-10-iMSCs-EV. Furthermore, from the RNA sequencing result, we could identify serval miRNA that accounted for the anti-inflammatory role in IL-10-iMSCs-EV. This combination has shown promise in several models of inflammatory diseases that IL-10-iMSCs were able to reduce inflammation more effectively than unmodified MSCs. The use of IL-10-iMSC-derived EVs further extends the therapeutic potential. Overall, the synergy between IL-10 and MSCs offers a promising approach to develop advanced therapies for inflammatory diseases to achieve better clinical outcomes.
