title: Suicidal gene therapy in an NF-κB-controlled tumor environment as monitored by a secreted blood reporter.
abstract: The nuclear factor-κB (NF-κB) is known to be activated in many cancer types including lung, ovarian, astrocytomas, melanoma, prostate as well as glioblastoma, and has been shown to correlate with disease progression. We have cloned a novel NF-κB-based reporter system (five tandem repeats of NF-κB responsive genomic element (NF; 14 bp each)) to drive the expression cassette for both a fusion between the yeast cytosine deaminase and uracil phosphoribosyltransferase (CU) as a therapeutic gene and the secreted Gaussia luciferase (Gluc) as a blood reporter, separated by an internal ribosomal entry site (NF-CU-IGluc). We showed that malignant tumor cells have high expression of Gluc, which correlates to high activation of NF-κB. When NF-κB was further activated by tumor necrosis factor-α in these cells, we observed up to 10-fold increase in Gluc levels and therefore transgene expression in human glioma cells served to greatly enhance the sensitization of these cells to the prodrug, 5-fluorocytosine both in cultured cells and in vivo subcutaneous tumor xenograft model. This inducible system provides a tool to enhance the expression of imaging and therapeutic genes for cancer therapy.
authors.affiliation: Department of Neurology, Neuroscience Center, Massachusetts General Hospital, Boston, MA 02129, USA.
authors.forename: C E
authors.initials: CE
authors.lastname: Badr
authors.forename: J M
authors.initials: JM
authors.lastname: Niers
authors.forename: D
authors.initials: D
authors.lastname: Morse
authors.forename: J A
authors.initials: JA
authors.lastname: Koelen
authors.forename: P
authors.initials: P
authors.lastname: Vandertop
authors.forename: D
authors.initials: D
authors.lastname: Noske
authors.forename: T
authors.initials: T
authors.lastname: Wurdinger
authors.forename: P A
authors.initials: PA
authors.lastname: Zalloua
authors.forename: B A
authors.initials: BA
authors.lastname: Tannous
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D004789:Enzyme Activation; D005437:Flucytosine / Q000378:metabolism; D041722:Genes, Transgenic, Suicide; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D066298:In Vitro Techniques; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D008819:Mice, Nude; D016328:NF-kappa B / Q000235:genetics* / Q000378:metabolism; D009368:Neoplasm Transplantation; D011401:Promoter Regions, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
chemical_list: D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; D005437:Flucytosine