세포 밖 소포를 모사한 나노 소포 제작 시스템 및 그 응용

Abstract

Extracellular vesicles (EVs) secreted from cells have potential uses as delivery agents for cell-free therapy. The key biological property of EVs is that they can deliver cellular materials of origin cells to target cells, and thereby stimulate them. Because of this feature, EVs have therapeutic effects on various diseases. However, cells produce EVs in small quantities. To overcome the difficulty of acquiring a sizable quantity of EVs at a time, we developed EV-mimetic nanovesicles called cell-engineered nanovesicles (CNVs) by breaking down the cells by extruding them through fine filters. We prove that CNVs and EVs are similar in properties and that CNVs are compatible with sufficient scale-up of production to be useful in clinical trials. Moreover, CNVs from embryonic stem cells can increase proliferation of mesenchymal stem cell (MSC) by delivering cellular materials and stimulating signal transduction to target MSC populations. To improve the ability of EV-mimetic nanovesicles to deliver cellular contents, I developed new EV-mimetic nanovesicles called bleb-derived nanovesicles (BNVs), and demonstrate that they have more membrane proteins and cytosolic proteins than EVs and CNVs to. Then I introduce a CRISPR/Cas9 gene editing system, into BNVs. I obtain Cas9-enriched vesicles from a cell in which the location of Cas9 is controlled, and demonstrate the potential of gene manipulation by BNVs. I demonstrate that EV-mimetic vesicles have various potential applications as nanocarriers. Hence, these results are expected to contribute to EV-related research and their various applications