Engineered bone marrow mesenchymal stem cell-derived exosomes loaded with miR302 through the cardiomyocyte specific peptide can reduce myocardial ischemia and reperfusion (I/R) injury

Abstract

Background MicroRNA (miRNA)-based therapies have shown great potential in myocardial repair following myocardial infarction (MI). MicroRNA-302 (miR302) has been reported to exert a protective effect on MI. However, miRNAs are easily degraded and ineffective in penetrating cells, which limit their clinical applications. Exosomes, which are small bioactive molecules, have been considered as an ideal vehicle for miRNAs delivery due to their cell penetration, low immunogenicity and excellent stability potential. Herein, we explored cardiomyocyte-targeting exosomes as vehicles for delivery of miR302 into cardiomyocyte to potentially treat MI. Methods To generate an efficient exosomal delivery system that can target cardiomyocytes, we engineered exosomes with cardiomyocyte specific peptide (CMP, WLSEAGPVVTVRALRGTGSW). Afterwards, the engineered exosomes were characterized and identified using transmission electron microscope (TEM) and Nanoparticle Tracking Analysis (NTA). Later on, the miR302 mimics were loaded into the engineered exosomes via electroporation technique. Subsequently, the effect of the engineered exosomes on myocardial ischemia and reperfusion (I/R) injury was evaluated in vitro and in vivo, including MTT, ELISA, real-time quantitative polymerase chain reaction (PCR), western blot, TUNNEL staining, echocardiogram and hematoxylin and eosin (HE) staining. Results Results of in vitro experimentation showed that DSPE-PEG-CMP-EXO could be more efficiently internalized by H9C2 cells than unmodified exosomes (blank‐exosomes). Importantly, compared with the DSPE-PEG-CMP-EXO group, DSPE-PEG-CMP-miR302-EXO significantly upregulated the expression of miR302, while exosomes loaded with miR302 could enhance proliferation of H9C2 cells. Western blot results showed that the DSPE-PEG-CMP-miR302-EXO significantly increased the protein level of Ki67 and Yap, which suggests that DSPE-PEG-CMP-miR302-EXO enhanced the activity of Yap, the principal downstream effector of Hippo pathway. In vivo, DSPE-PEG-CMP-miR302-EXO improved cardiac function, attenuated myocardial apoptosis and inflammatory response, as well as reduced infarct size significantly. Conclusion In conclusion, our findings suggest that CMP-engineered exosomes loaded with miR302 was internalized by H9C2 cells, an in vitro model for cardiomyocytes coupled with potential enhancement of the therapeutic effects on myocardial I/R injury.

Emerging Strategies in Mesenchymal Stem Cell-Based Cardiovascular Therapeutics

“…In the context of cardiovascular diseases, MSC-derived EVs exhibit potential in enhancing angiogenesis, reducing fibrosis, and modulating inflammatory responses, which are crucial for myocardial repair and regeneration [46]. The therapeutic efficacy of EVs is largely attributed to their ability to mirror the regenerative and reparative properties of their parent MSCs while offering an advantage in terms of safety and feasibility as they lack the risks associated with stem cell implantation, such as tumorigenicity and immune rejection [47]. This study has investigated the use of cardiomyocyte-targeting exosomes loaded with microRNA-302 (miR302) for treating myocardial ischemia/reperfusion injury, finding that these engineered exosomes enhanced cardiomyocyte proliferation and activity and reduced heart damage in vitro and in mice [47].…”

“…The therapeutic efficacy of EVs is largely attributed to their ability to mirror the regenerative and reparative properties of their parent MSCs while offering an advantage in terms of safety and feasibility as they lack the risks associated with stem cell implantation, such as tumorigenicity and immune rejection [47]. This study has investigated the use of cardiomyocyte-targeting exosomes loaded with microRNA-302 (miR302) for treating myocardial ischemia/reperfusion injury, finding that these engineered exosomes enhanced cardiomyocyte proliferation and activity and reduced heart damage in vitro and in mice [47]. Recent advances have focused on optimizing the isolation, characterization, and scalability of EV production under good manufacturing practice (GMP) conditions [48].…”

Advances in Nanoparticles in the Prevention and Treatment of Myocardial Infarction

“…Myocardial infarction (ischemia-reperfusion, mice) Ejection fraction and other indicators were improved and ventricular remodeling was reduced [121] Table 3. Cont.…”