Targeted expression of green fluorescent protein/tumor necrosis factor-related apoptosis-inducing ligand fusion protein from human telomerase reverse transcriptase promoter elicits antitumor activity without toxic effects on primary human hepatocytes.
Liver toxicity is the major concern for use of recombinant human tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) proteins in treatment of cancers. Here we report that normal human primary hepatocytes (NHPHs) are susceptible to the transduction of the wild-type, full-length coding sequence of the human TRAIL gene. To minimize potential toxicity of the TRAIL gene, a bicistronic adenoviral vector that expresses the green fluorescent protein/TRAIL fusion protein from the human telomerase reverse transcriptase promoter (designated Ad/gTRAIL) was constructed. In vitro and in vivo studies have showed that treatment with the adenoviral vector Ad/gTRAIL results in high-level expression of green fluorescent protein/TRAIL in cancer cells but no detectable transgene expression in NHPHs or in normal mouse liver tissues. Furthermore, treatment with Ad/gTRAIL effectively elicited apoptosis in malignant cells but not in NHPHs in vitro and suppressed tumor growth and prolonged duration of survival in vivo. Thus, with the combined advantages of the TRAIL gene and the human telomerase reverse transcriptase target, Ad/gTRAIL can be a potent therapeutic agent for the treatment of cancers.
Lin
Tongyu
T
Department of Thoracic and Cardiovascular Surgery, The University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030, USA.
Gu
Jian
J
Zhang
Lidong
L
Huang
Xuefeng
X
Stephens
L Clifton
LC
Curley
Steven A
SA
Fang
Bingliang
B
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D051017:Apoptosis Regulatory Proteins; D002455:Cell Division / Q000502:physiology; D003110:Colonic Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D004268:DNA-Binding Proteins; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D008164:Luminescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D008175:Lung Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000235:genetics / Q000633:toxicity; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D053221:TNF-Related Apoptosis-Inducing Ligand; D019098:Telomerase / Q000235:genetics*; D019076:Transgenes; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis* / Q000235:genetics / Q000633:toxicity; D023041:Xenograft Model Antitumor Assays
D051017:Apoptosis Regulatory Proteins; D004268:DNA-Binding Proteins; D008164:Luminescent Proteins; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; D053221:TNF-Related Apoptosis-Inducing Ligand; C505653:TNFSF10 protein, human; C505654:Tnfsf10 protein, mouse; D014409:Tumor Necrosis Factor-alpha; D049452:Green Fluorescent Proteins; D019098:Telomerase