Development and characterization of recombinant adenoviruses encoding human p53 for gene therapy of cancer.
We have constructed recombinant human adenoviruses that express wild-type human p53 under the control of either the Ad 2 major late promoter (MLP) or the human cytomegalovirus (CMV) immediate early gene promoter. Each construct replaces the Ad 5 E1a and E1b coding sequences necessary for viral replication with the p53 cDNA and MLP or CMV promoter. These p53/Ad recombinants are able to express p53 protein in a dose-dependent manner in infected human cancer cells. Tumor suppressor activity of the expressed p53 protein was assayed by several methods. [3H]Thymidine incorporation assays showed that the recombinant adenoviruses were capable of inhibiting DNA synthesis in a p53-specific, dose-dependent fashion. Ex vivo treatment of Saos-2 tumor cells, followed by injection of the treated cells into nude mice, led to complete tumor suppression using the MLP/p53 recombinant. Following a single injection of CMV/p53 recombinant adenovirus into the peritumoral space surrounding an in vivo established tumor derived from a human small cell lung carcinoma cell line (NIH-H69), we were able to detect p53 mRNA in the tumors at 2 and 7 days post-injection. Continued treatment of established H69 tumors with MLP/p53 recombinant led to reduced tumor growth and increased survival time compared to control treated animals. These results indicate that recombinant adenoviruses expressing wild-type p53 may be useful vectors for gene therapy of human cancer.
Wills
K N
KN
CANJI, Inc., San Diego, CA 92121.
Maneval
D C
DC
Menzel
P
P
Harris
M P
MP
Sutjipto
S
S
Vaillancourt
M T
MT
Huang
W M
WM
Johnson
D E
DE
Anderson
S C
SC
Wen
S F
SF
D000260:Adenoviruses, Human / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D018288:Carcinoma, Small Cell / Q000473:pathology / Q000628:therapy*; D003587:Cytomegalovirus / Q000235:genetics; D004261:DNA Replication; D018076:DNA, Complementary / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D003673:Defective Viruses / Q000235:genetics*; D005260:Female; D016158:Genes, p53; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D008175:Lung Neoplasms / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008969:Molecular Sequence Data; D009368:Neoplasm Transplantation; D009369:Neoplasms / Q000235:genetics / Q000628:therapy*; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000172:deficiency / Q000235:genetics
D018076:DNA, Complementary; D004274:DNA, Recombinant; D004279:DNA, Viral; D011993:Recombinant Fusion Proteins; D016159:Tumor Suppressor Protein p53