Naturally derived nanovesicles secreted from various cell types and found in body fluids can provide effective platforms for the delivery of various cargoes because of their intrinsic ability to be internalized for intercellular signal transmission and membrane recycling. In this study, the versatility of bioengineered extracellular membranous nanovesicles as potent carriers of small-interfering RNAs (siRNAs) for stem cell engineering and in vivo delivery has been explored. Here, exosomes have been engineered, one of the cell-derived vesicle types, to overexpress exosomal proteins fused with cell-adhesion or cell-penetrating peptides for enhanced intracellular gene transfer. To devise a more effective delivery system with potential for mass production, a new siRNA delivery system has also been developed by artificially inducing the outward budding of plasma membrane nanovesicles. Those nanovesicles have been engineered by overexpressing E-cadherin to facilitate siRNA delivery to human stem cells with resistance to intracellular gene transfer. Both types of engineered nanovesicles deliver siRNAs to human stem cells for lineage specification with negligible cytotoxicity. The nanovesicles are efficient in delivering siRNA in vivo, suggesting feasibility for gene therapy. Cell-derived, bioengineered nanovesicles used for siRNA delivery can provide functional platforms enabling effective stem cell therapeutics and in vivo gene therapy.
Bibliographical noteFunding Information:
Y.J. and J.S.L. contributed equally to this work. This work was supported by grants (NRF-2013R1A1A2A10061422, 2015M3C9A4053251, and 2015R1A2A1A15053771) from the National Research Foundation (NRF) of Korea, and a grant from the Translational Research Center for Protein Function Control (TRCP 2016R1A5A1004694) funded by the National Research Foundation (NRF) of Korea. This work was also supported in part by the BK21 PLUS program. Human stem cells (hiPSCs and hNSCs) were kindly provided from Yonsei University College of Medicine with the approval of Institutional Review Board (IRB) of Yonsei University. All animal experiments were performed in accordance with the Korean Food and Drug Administration guidelines. Experimental protocols for in vivo imaging and siRNA delivery were reviewed and approved by the Yonsei Laboratory Animal Research Center (YLARC).
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All Science Journal Classification (ASJC) codes
- Materials Science(all)
- Condensed Matter Physics