Δ24-hyCD adenovirus suppresses glioma growth in vivo by combining oncolysis and chemosensitization

被引:45
作者
Conrad, C
Miller, CR
Ji, YJ
Gomez-Manzano, C
Bharara, S
McMurray, JS
Lang, FF
Wong, F
Sawaya, R
Yung, WKA
Fueyo, J
机构
[1] Univ Texas, MD Anderson Canc Ctr, Dept Neurooncol, Brain Tumor Ctr, Houston, TX 77030 USA
[2] Univ Texas, MD Anderson Canc Ctr, Dept Neurosurg, Brain Tumor Ctr, Houston, TX 77030 USA
[3] Dept Nucl Imaging, Houston, TX USA
[4] Univ Alabama, Sch Med, Birmingham, AL USA
[5] Washington Univ, Sch Med, Dept Pathol & Immunol, St Louis, MO USA
关键词
oncolytic virus; Delta; 24; cytosine deaminase; suicide gene;
D O I
10.1038/sj.cgt.7700750
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Replication-competent adenoviruses could provide an efficient method for delivering therapeutic genes to tumors. The most promising strategies among adenovirus-based oncolytic systems are designed to exploit free E2F-1 activity in cancer cells, which in the absence of pRb activates transcription and regulates the expression of genes involved in differentiation, proliferation, and apoptosis. We previously developed Delta24, an E1A-mutant, conditionally replicative oncolytic adenovirus. Here, we examine the ability of a second-generation Delta24 (Delta24-hyCD) engineered to express a humanized form of the Saccharomyces cerevisiae cytosine deaminase gene (hyCD). Real-time quantitative PCR, Western blotting, thin-layer chromatography, and radioisotope quantitative enzymatic assays confirmed the production of a catalytically active hyCD enzyme in the setting of an oncolytic infection in vitro; other experiments assessing local production of 5-fluorouracil and a concomitant bystander effect showed improved cytotoxicity. The IC50 dose of 5-fluorocytosine (5-FC) required for a complete cytopathic effect by the Delta24-hyCD virus was fivefold lower than with Delta24 alone in U251MG and U87MG malignant glioma (MG) cell lines. Intratumoral treatment of mice bearing intracranial U87MG xenografts with Delta24-hyCD + 5-FC significantly improved survival, confirming that Delta24-hyCD with 5-FC is a more efficient anticancer tool than Delta24 alone. Histopathologically, Delta24-hyCD replication was accompanied by progressively augmented oncolysis and drug-induced necrosis. These findings demonstrate that Delta24-hyCD with concomitant systemic 5-FC is a significant improvement over the earlier Delta24 oncolytic tumor-selective strategy for therapy of experimental gliomas.
引用
收藏
页码:284 / 294
页数:11
相关论文
共 27 条
[1]  
Aghi M, 1999, CANCER RES, V59, P3861
[2]   Gene therapy for gliomas:: Molecular targets, adenoviral vectors, and oncolytic adenoviruses [J].
Alemany, R ;
Gomez-Manzano, C ;
Balagué, C ;
Yung, WKA ;
Curiel, DT ;
Kyritsis, AP ;
Fueyo, J .
EXPERIMENTAL CELL RESEARCH, 1999, 252 (01) :1-12
[3]  
Bernt KM, 2002, CANCER RES, V62, P6089
[4]   An adenovirus mutant that replicates selectively in p53-deficient human tumor cells [J].
Bischoff, JR ;
Kim, DH ;
Williams, A ;
Heise, C ;
Horn, S ;
Muna, M ;
Ng, L ;
Nye, JA ;
SampsonJohannes, A ;
Fattaey, A ;
McCormick, F .
SCIENCE, 1996, 274 (5286) :373-376
[5]  
Blanquicett C, 2002, MOL CANCER THER, V1, P1139
[6]  
BOVIATSIS EJ, 1994, CANCER RES, V54, P5745
[7]  
*CENTR BRAIN TUM R, 2001, STAT REP PRIM BRAIN
[8]   An oncolytic viral mutant that delivers the CYP2B1 transgene and augments cyclophosphamide chemotherapy [J].
Chase, M ;
Chung, RY ;
Chiocca, EA .
NATURE BIOTECHNOLOGY, 1998, 16 (05) :444-448
[9]   Characterization of the Saccharomyces cerevisiae FCY1 gene encoding cytosine deaminase and its homologue FCA1 of Candida albicans [J].
Erbs, P ;
Exinger, F ;
Jund, R .
CURRENT GENETICS, 1997, 31 (01) :1-6
[10]  
Freytag SO, 2003, CANCER RES, V63, P7497