Low-frequency ultrasound increases non-viral gene transfer to the mouse lung  

作　　者：Stefania Xenariou Hai-Dong Liang Uta Griesenbach Jie Zhu Raymond Farley Lucinda Somerton Charanjit Singh Peter K. Jeffery Ronald K. Scheule Seng H. Cheng Duncan M. Geddes Martin Blomley Eric W. F. W. Alton 

机构地区：[1]Department of Gene Therapy, National Heart and Lung Institute, Faculty of Medicine, Imperial College, London, UK [2]UK Cystic Fibrosis Gene Therapy Consortium, London, UK [3]Ultrasound Group, Imaging Sciences Department, MRC Clinical Sciences Centre, Imperial College, Faculty of Medicine, London, UK [4]Genzyme Corporation, Framingham, MA, USA [5]Present address: School of Engineering, Cardiff University, Cardiff, UK

出　　处：《Acta Biochimica et Biophysica Sinica》2010年第1期45-51,共7页生物化学与生物物理学报（英文版）

摘　　要：The aim of the study was to assess if low-frequency ultrasound （US）, in the range of 30-35 kHz, increases non-viral gene transfer to the mouse lung. US is greatly attenuated in the lung due to large energy losses at the air/tissue interfaces. The advantages of low-frequency US, compared with high-frequency US are： （i） increased cavi- tation （responsible for the formation of transient pores in the cell membrane） and （ii） reduced energy losses during lung penetration. Cationic lipid GL67/plasmid DNA （pDNA）, polyethylenimine （PEI）/pDNA and naked pDNA were delivered via intranasal instillation and the animals were then exposed to US （sonoporation） at 0.07 or 0.1 MPa for 10 rain. Under these conditions, US did not enhance GL67 or PEI-mediated transfection. It did, however, increase naked pDNA gene transfer by approxi- mately 4 folds. Importantly, this was achieved in the absence of microbubbles, which are crucial for the com- monly used high-frequency （1 MHz） sonoporation but may not be able to withstand nebulization in a clinically relevant setup. Lung hemorrhage was also assessed and shown to increase with US pressure in a dose-dependent manner. We have thus, established that low-frequency US can enhance lung gene transfer with naked pDNA and this enhancement is more effective than the previously reported 1 MHz US.The aim of the study was to assess if low-frequency ultrasound （US）, in the range of 30-35 kHz, increases non-viral gene transfer to the mouse lung. US is greatly attenuated in the lung due to large energy losses at the air/tissue interfaces. The advantages of low-frequency US, compared with high-frequency US are： （i） increased cavi- tation （responsible for the formation of transient pores in the cell membrane） and （ii） reduced energy losses during lung penetration. Cationic lipid GL67/plasmid DNA （pDNA）, polyethylenimine （PEI）/pDNA and naked pDNA were delivered via intranasal instillation and the animals were then exposed to US （sonoporation） at 0.07 or 0.1 MPa for 10 rain. Under these conditions, US did not enhance GL67 or PEI-mediated transfection. It did, however, increase naked pDNA gene transfer by approxi- mately 4 folds. Importantly, this was achieved in the absence of microbubbles, which are crucial for the com- monly used high-frequency （1 MHz） sonoporation but may not be able to withstand nebulization in a clinically relevant setup. Lung hemorrhage was also assessed and shown to increase with US pressure in a dose-dependent manner. We have thus, established that low-frequency US can enhance lung gene transfer with naked pDNA and this enhancement is more effective than the previously reported 1 MHz US.

关 键 词：low-frequency ultrasound SONOPORATION gene transfer non-viral vector LUNG cystic fibrosis 

分 类 号：S855.3[农业科学—临床兽医学] TD452[农业科学—兽医学]