Extracellular Vesicles: Engineered Drug Delivery Vehicles to the Cancer Cells!
Background
Multiple myeloma (MM) is a plasma cell malignancy that starts from a pre-malignant stage (MGUS) and develops into MM. This progression strictly depends on the bone marrow microenvironment (BMM). Recently our group showed that MM cell-derived Extracellular Vesicles (EVs) deliver molecular signals within the BMM affecting cell behaviour. EVs can be used to deliver cargo, this makes them promising vehicles for therapeutic agents. We aim to provide the proof-of-concept for a siRNA-mediated approach using EVs as a drug delivery tool.
Methods
To study the specificity of EVs’ delivery to tumor, we performed an in vivo experiment using a xenograft C57BL/6 mice MM model. Next, to apply EVs as a drug delivery system, I used sonication for loading siRNA/miRNA into MM-EVs and evaluated: I) EVs number and size profile by Nanosight and their uptake in MM cells by FACs before and after sonication; II) The loading efficacy of a synthetic miRNA (cel-39), which is easily detectable in human EVs and cells using a TaqMan assay; III) The molecular effect after the uptake of siRNA-loaded EVs by MM cell through qPCR analysis.
Results
The injection of EVs in the xenograft MM model shows that EVs can be used as a specific siRNA delivery tool. The analysis of the sonicated EVs indicated that sonication is a safe method. In addition, the results of cel-39 assay suggested that sonication could be effective for delivering siRNA/miRNA into EVs. Moreover, the preliminary results of the functional assay showed that miRNA engineered-vesicles can effectively deliver an inhibitory signal in the target cells.
Conclusion
In conclusion, I defined a protocol of sonication suitable for engineering EVs that can deliver inhibitory signals to cancer cells through sRNA molecules. Although this preliminary protocol still needs to be optimized, the present results indicate that EVs are a promising drug delivery system for anti-cancer therapy.
Keywords
Multiple myeloma- MM; Extracellular vesicles-EVs; drug delivery, siRNA; miRNA
Funding/Acknowledgments
This study was supported by grants from Associazione Italiana Ricerca sul Cancro, AIRC Investigator Grant to RC (20614) and Intersectoral Innovation PhD program to NA. A special thanks to my colleagues Domenica Giannandrea, Valentina Citro, Lavinia Casati, Alessandro Villa, who helped me with the progression of the experiments and professor Raffaella Chiaramonte who supported me along the project.
Authors
Nazanin Abazari1 (Corresponding Author: Nazanin.Abazari[at]unimi[dot]it), Domenica Giannandrea1, Valentina Citro1, Lavinia Casati1, Natalia Platonova1, Giulia Sauro1, Ehsan Soleymaninejadian1, Lara Paonessa1, Jad Kabbout1, Alessandro Villa1, Elisabetta Crippa1, Paolo Ciana1, Raffaella Chiaramonte1