Liposome-mediated NGF gene transfection following neuronal injury: potential therapeutic applications.
We have systematically investigated the therapeutic potential of cationic liposome-mediated neurotrophic gene transfer for treatment of CNS injury. Following determination of optimal transfection conditions, we examined the effects of dimethylaminoethane-carbamoyl-cholesterol (DC-Chol) liposome-mediated NGF cDNA transfection in injured and uninjured primary septo-hippocampal cell cultures and rat brains. In in vitro studies, we detected an increase of NGF mRNA in cultures 1 day after transfection. Subsequent ELISA and PC12 cell biological assays confirmed that cultured cells secreted soluble active NGF into the media from day 2 after gene transfection. Further experiments showed that such NGF gene transfection reduced the loss of chol- ine acetyltransferase (ChAT) activity in cultures following calcium-dependent depolarization injury. In in vivo studies, following intraventricular injections of NGF cDNA complexed with DC-Chol liposomes, ELISA detected nine- to 12-fold increases of NGF in rat CSF. Further studies showed that liposome/NGF cDNA complexes could attenuate the loss of cholinergic neuronal immunostaining in the rat septum after traumatic brain injury (TBI). Since deficits in cholinergic neurotransmission are a major consequence of TBI, our studies demonstrate for the first time that DC-Chol liposome-mediated NGF gene transfection may have therapeutic potential for treatment of brain injury.
Zou
L L
LL
Department of Neurosurgery, Baylor College of Medicine, Houston, TX 77030, USA.
Huang
L
L
Hayes
R L
RL
Black
C
C
Qiu
Y H
YH
Perez-Polo
J R
JR
Le
W
W
Clifton
G L
GL
Yang
K
K
D000818:Animals; D001930:Brain Injuries / Q000201:enzymology / Q000628:therapy*; D002478:Cells, Cultured; D002795:Choline O-Acetyltransferase / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods*; D006624:Hippocampus; D008081:Liposomes; D009414:Nerve Growth Factors / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection / Q000235:genetics*
D008081:Liposomes; D009414:Nerve Growth Factors; D002795:Choline O-Acetyltransferase