Tumor-microenvironment Responsive Targeted Exosome-like Nanovesicles From Dunaliella Salina for Enhancing Gene/immune Combination Therapy

Abstract BackgroundAs an endogenous extracellular vesicle, exosome is increasingly presenting its great potential in the field of drug delivery. However, it is the bottleneck to obtain a large number of uniform, stable and multi-component controllable exosomes with low cost and time. ResultsIn this study, we develop a novel targeted drug delivery system based on exosome-like nanovesicle by use of natural marine single-celled Dunaliella salina (DENV), the c(RGDyK) peptide has been conjugated to DENV surface to achieve the targeted delivery to esophageal cancer cells. Furthermore, miR-375 has been loaded into the cRGD-DENV through electroporation, and aPD-L1 has been conjugated onto its surface via Gly-PLGLAG-Cys peptide, a matrix metalloproteinase-2 (MMP-2)-cleavable peptide, which facilitates the release of aPD-L1 in tumor environment to achieve the high-efficiency combination of gene therapy and immunotherapy. Firstly, the engineered DENV delivery system was prepared and characterized. It exhibited a proper particle diameter (approximately 150 nm) with in vitro sustained release features in the presence of MMP-2/9. More importantly, the cRGD-DENV was effective, promoted selective delivery of cargo to the target site, and reduced nonspecific uptake, consequently, significantly inhibit tumor growth in vitro and in vivo. ConclusionThe specific nanocarrier delivery system provide a promising strategy for the rapid and large-scale production of functionalized exosome-like nanovesicle by adapting multifunctional peptides specifically targeted tumor.