Gene transfer into adult rat spinal cord using naked plasmid DNA and ultrasound microbubbles.

Although gene therapy might become a promising approach to treat spinal cord injury, the safety issue is a serious consideration in human gene therapy. Plasmid DNA transfer is safer than viral vectors, but the transfection efficiency is quite low. To overcome the problem, we applied the ultrasound microbubbles-mediated transfection method to the spinal cord in adult rats, since ultrasound microbubbles have been reported to be efficient to increase transfection efficiency in various tissues. After exposing T9-10 spinal cord with a laminectomy, we injected a mixture of naked plasmid DNA and microbubbles into cerebrospinal fluid by lumbar puncture. Then, the T9-10 spinal cord was exposed to ultrasound. An ultrasound intensity of 0.4-0.5 W/cm2 significantly increased luciferase expression up to approximately 15-60-fold at the insonated level as compared to naked plasmid DNA alone. Luciferase activity could be detected at least up to 7 days after transfection, while the expression level was almost returned to undetectable level at 14 days after transfection. The transfected cells were mainly meningeal cells in the surface of insonated spinal cord. There was no obvious evidence of worsening of neurological deficits as compared to rats transfected with naked plasmid DNA alone or untransfected rats. Similarly, successful gene transfer was also achieved in the insonated T9-10 spinal cord after spinal cord injury. Overall, the present study demonstrated the feasibility of ultrasound microbubbles-mediated plasmid DNA transfer into the target level of the spinal cord.

Shimamura M ，Sato N ，Taniyama Y ，Kurinami H ，Tanaka H ，Takami T ，Ogihara T ，Tohyama M ，Kaneda Y ，Morishita R ... -  《JOURNAL OF GENE MEDICINE》

Sustained transgene expression in intervertebral disc cells in vivo mediated by microbubble-enhanced ultrasound gene therapy.

Nishida K ，Doita M ，Takada T ，Kakutani K ，Miyamoto H ，Shimomura T ，Maeno K ，Kurosaka M ... -  《-》

Development of safe and efficient novel nonviral gene transfer using ultrasound: enhancement of transfection efficiency of naked plasmid DNA in skeletal muscle.

Taniyama Y ，Tachibana K ，Hiraoka K ，Aoki M ，Yamamoto S ，Matsumoto K ，Nakamura T ，Ogihara T ，Kaneda Y ，Morishita R ... -  《GENE THERAPY》

Development of efficient plasmid DNA transfer into adult rat central nervous system using microbubble-enhanced ultrasound.

Shimamura M ，Sato N ，Taniyama Y ，Yamamoto S ，Endoh M ，Kurinami H ，Aoki M ，Ogihara T ，Kaneda Y ，Morishita R ... -  《GENE THERAPY》

Spinal gene transfer using ultrasound and microbubbles.

Takahashi M ，Kido K ，Aoi A ，Furukawa H ，Ono M ，Kodama T ... -  《JOURNAL OF CONTROLLED RELEASE》