Recombinant baculovirus containing the diphtheria toxin A gene for malignant glioma therapy.
Insect baculoviruses are capable of infecting mammalian glial cells in the central nervous system. We investigated in the current study the feasibility of using the viruses as toxin gene vectors to eliminate malignant glioma cells in the brain. We first confirmed that glioma cells were permissive to baculovirus infection, with variable transduction efficiencies at 100 viral particles per cell and ranging between 35% and 70% in seven human and rat glioma cell lines. We then developed a recombinant baculovirus vector accommodating the promoter of glial fibrillary acidic protein (GFAP) to minimize possible side effects caused by overexpression of a therapeutic gene in sensitive neurons. We placed the GFAP promoter into a baculovirus expression cassette, in which the enhancer of human cytomegalovirus immediate-early gene and the inverted terminal repeats of adeno-associated virus were employed to improve the relatively low transcriptional activity of the cellular promoter. This recombinant baculovirus significantly improved transduction in glioma cells, providing the efficiency in C6 rat glioma cells up to 96%. When used to produce the A-chain of diphtheria toxin intracellularly in a rat C6 glioma xenograft model, the baculovirus effectively suppressed tumor development. The new baculovirus vector circumvents some of the inherent problems associated with mammalian viral vectors and provides an additional option for cancer gene therapy.
Wang
Chao-Yang
CY
Institute of Bioengineering and Nanotechnology and Department of Biological Sciences, National University of Singapore, Singapore.
Li
Feng
F
Yang
Yi
Y
Guo
Hai-Yan
HY
Wu
Chun-Xiao
CX
Wang
Shu
S
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D001932:Brain Neoplasms / Q000235:genetics* / Q000628:therapy*; D048708:Cell Growth Processes / Q000235:genetics; D045744:Cell Line, Tumor; D004167:Diphtheria Toxin / Q000096:biosynthesis / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D005910:Glioma / Q000235:genetics* / Q000628:therapy* / Q000821:virology; D006801:Humans; D008297:Male; D051379:Mice; D041681:NIH 3T3 Cells; D009363:Neoplasm Proteins / Q000096:biosynthesis; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics*; D011401:Promoter Regions, Genetic; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014161:Transduction, Genetic; D019076:Transgenes; D023041:Xenograft Model Antitumor Assays
D004167:Diphtheria Toxin; D009363:Neoplasm Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; C029795:diphtheria toxin fragment A