Development of synthetic promoters for radiation-mediated gene therapy

HSV Recombinant Vectors for Gene Therapy

“…A limitation in the use of these promoters is that viral replication is mainly targeted at a specific tumour type and often further restricted to only a subgroup. To this purpose, promoters active in most tumours [18, 247, 262] have been explored [263, 264], or radiation-responsive promoters, but there are instances where these promoters might be active in normal cells, leading to toxicity. A further obstacle is that, in the context of a tumour, it is likely that these promoters will not be active in all tumour cells and that not all cells will be infected.…”

Gene therapy vectors containing CArG elements from the Egr1 gene are activated by neutron irradiation, cisplatin and doxorubicin

“…[16][17][18] Cell culture and transfection Human MCF-7 breast adenocarcinoma and U373-MG glioblastoma cells were obtained from the European Collection of Cell Cultures (Salisbury, UK). Cells were maintained in monolayer culture in complete RPMI medium (Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin, 100 mg/ml streptomycin (Sigma Aldrich, St. Louis, MO), 2 mM L-glutamine (Invitrogen), and incubated in a humidified incubator at 371C in 5% O 2 :5% CO 2 :90% N 2 .…”

“…Transient transfections with the GFP-constructs were carried out by using Lipofectamine (Invitrogen), as described previously. 16 Transient transfections with the HSVtk-constructs were performed using TransIT (Mirus Corporation, Madison, WI), according to manufacturer's instructions.…”

“…15 In an attempt to produce more responsive promoters and to eliminate binding sites for nonradiation-induced transcription factors, we have produced synthetic promoters containing tandem repeats of isolated CArG elements. 16 These new promoters demonstrated greater inducibility and lower basal activity than the wild-type Egr1 promoter in a number of human tumor cell types, 16,17 and could efficiently control experimental suicide gene therapy after clinically significant doses of X-rays in vitro and in vivo. [16][17][18][19] Gene therapy vectors containing CArG elements also have potential to be used in combination with chemotherapy, since the native Egr1 gene has been shown to be induced by cisplatin 20,21 and the anthracycline antibiotic doxorubicin.…”

“…16 These new promoters demonstrated greater inducibility and lower basal activity than the wild-type Egr1 promoter in a number of human tumor cell types, 16,17 and could efficiently control experimental suicide gene therapy after clinically significant doses of X-rays in vitro and in vivo. [16][17][18][19] Gene therapy vectors containing CArG elements also have potential to be used in combination with chemotherapy, since the native Egr1 gene has been shown to be induced by cisplatin 20,21 and the anthracycline antibiotic doxorubicin. 22 ''Chemogenetic'' therapy may represent a novel and promising avenue for combinational therapy, since the efficacy of these antineoplastic agents is often limited by normal tissue toxicity and tumor resistance.…”