Biocompatible polymer enhances the in vitro and in vivo transfection efficiency of HVJ envelope vector.
BACKGROUND
Vector development is critical for the advancement of human gene therapy. However, the use of viral vectors raises many safety concerns and most non-viral methods are less efficient for gene transfer. One of the breakthroughs in vector technology is the combination of the vector with various polymers.


METHODS
HVJ (hemagglutinating virus of Japan) envelope vector (HVJ-E) has been developed as a versatile gene transfer vector. In this study, we combined HVJ-E with cationized gelatin to make it a more powerful tool and assessed its transfection efficiency in vitro and in vivo. In addition, we investigated the mechanism of the gene transfer by means of the inhibition of fusion or endocytosis.


RESULTS
The combination of both protamine sulfate and cationized gelatin with HVJ-E, referred to as PS-CG-HVJ-E, further enhanced the in vitro transfection efficiency. In CT26 cells, the luciferase gene expression of PS-CG-HVJ-E was approximately 10 times higher than that of the combination of protamine sulfate with HVJ-E or the combination of cationized gelatin with HVJ-E, referred to as PS-HVJ-E or CG-HVJ-E, respectively. Furthermore, the luciferase gene expression in liver mediated by intravenous administration of CG-HVJ-E was much higher than the luciferase gene expression mediated by PS-HVJ-E or PS-CG-HVJ-E and approximately 100 times higher than that mediated by HVJ-E alone.


CONCLUSIONS
Cationized gelatin-conjugated HVJ-E enhanced gene transfection efficiency both in vitro and in vivo. These results suggest that low molecular weight cationized gelatin may be appropriate for complex formation with various envelope viruses, such as retrovirus, herpes virus and HIV.
Mima
Hidetoshi
H
Division of Gene Therapy Science, Graduate School of Medicine, Osaka University, 2-2 Yamada-oka, Suita, Osaka 565-0871, Japan.
Tomoshige
Ryuji
R
Kanamori
Toshihide
T
Tabata
Yasuhiko
Y
Yamamoto
Seiji
S
Ito
Susumu
S
Tamai
Katsuto
K
Kaneda
Yasufumi
Y
D000730:Androstadienes; D000818:Animals; D001672:Biocompatible Materials; D002412:Cations; D002460:Cell Line; D005780:Gelatin; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D011108:Polymers; D029082:Sendai virus / Q000235:genetics*; D014162:Transfection / Q000379:methods*; D000077191:Wortmannin
D000730:Androstadienes; D001672:Biocompatible Materials; D002412:Cations; D011108:Polymers; D005780:Gelatin; D000077191:Wortmannin