Yusuke OdJohn Ung Diki OmtHitoshi UrugYoichi Negishi,Kzuo Mruym

aFaculty of Pharma-Sciences,Teikyo University,Tokyo,Japan

bSchool of Pharmacy,Tokyo University of Pharmacy and Life Science,Tokyo,Japan

Development of cell-targeting bubble liposome for effective gene delivery with ultrasound

Mutsumi Sugiia,*,Ryo Suzukia,Yusuke Odaa,Johan Ungaa, Daiki Omataa,Hitoshi Urugaa,Yoichi Negishib,Kazuo Maruyamaa

aFaculty of Pharma-Sciences,Teikyo University,Tokyo,Japan

bSchool of Pharmacy,Tokyo University of Pharmacy and Life Science,Tokyo,Japan

A R T I C L E I N F O

Article history:

Available online 25 November 2015

Cell-targeting bubble liposome

Gene delivery

Ultrasound

Previously,we developed a gene delivery system using Bubble liposomes(perfuoropropane gas-entrapping liposomes;BLs) and ultrasound(US)[1].The combination of BLs and US can deliver plasmid DNA into the cytoplasm via the formation of transient membrane pores by cavitation.To enhance the effciency of gene delivery,it is important to induce the cavitation of BLs at the vicinity of cells.In brief,cell-binding BLs would be useful for gene delivery to induce the cavitation on the cell membrane.In this study,we prepared the BL conjugated cyclic RGD(cRGD)peptides which could bind to tumor neovessel[2], and we investigated gene transfection effciency for Human Umbilical Vein Endothelial Cells(HUVECs)with cRGD conjugated BLs(cRGD-BLs)and US.

Binding assay:Fluorescence labeled cRGD-BLs were incubated with HUVECs at 4°C.After 1 hr,the cells were washed and the fuorescence intensity was measured with fowcytometer.Gene delivery:Luciferase coded plasmid DNA(pCMVLuc)and either cRGD-BLs or BLs(60 μg/mL)were added to HUVECs.Then,US(2 MHz,2.5 W/cm2,10 sec.)was exposed.After that,HUVECs were washed with PBS and incubated for 24 hr. Finally,luciferase expression was measured.cRGD-BLs effectively bound to HUVECs compared to non-targeted BLs. Luciferase expression in the group of treatment with cRGDBLs and US was higher than that with BLs and US(Fig.1).This result suggested that cRGD-BLs effectively delivered pCMVLuc into HUVECs.Although the mechanism of enhancement for gene delivery effciency is not clear,it is thought that many transient pores as a route of gene delivery might be opened by the cavitation of cRGD-BLs after binding on cell membrane.We found out that cRGD-BLs could enhance the gene delivery effciency with US.Therefore,the combination of celltargeting BLs and US would be an effective gene delivery system.

Acknowledgements

This study was supported by MEXT-supported Program for the Strategic Research Foundation at Private Universities,2013–2017.

Fig.1–Luciferase activity in HUVECs transfected with pCMV-Luc.

R E F E R E N C E S

[1]Suzuki R.Effective gene delivery with liposomal bubbles and ultrasound as novel non-viral system.J Drug Target 2007;7-8:531–537.

[2]Hynes RO.A reevaluation of integrins as regulators of angiogenesis.Nat Med 2001;8:918–921.

*E-mail address:seki@pharm.teikyo.u-ac.jp.

Peer review under responsibility of Shenyang Pharmaceutical University.

http://dx.doi.org/10.1016/j.ajps.2015.11.083

1818-0876/?2016 Production and hosting by Elsevier B.V.on behalf of Shenyang Pharmaceutical University.This is an open access article under the CC BY-NC-ND license(http://creativecommons.org/licenses/by-nc-nd/4.0/).