Chimeric tumor suppressor 1, a p53-derived chimeric tumor suppressor gene,kills p53 mutant and p53 wild-type glioma cells in synergy with irradiation and CD95 ligand

Adenoviral chimeric tumor suppressor I (CTS1) gene transfer was evaluated a s a novel approach of somatic gene therapy for malignant glioma. CTS1 is an artificial p53-based gene designed to resist various pathways of p53 inact ivation. Here, we report that an adenovirus encoding CTS1 (Ad-CTS1) induces growth arrest and loss of viability in all glioma cell lines examined, in the absence of specific cell cycle changes. In contrast, an adenovirus enco ding wild-type p53 (Ad-p53) does not consistently induce apoptosis in the s ame cell lines. Electron microscopic analysis of Ad-CTS1-infected glioma ce lls reveals complex cytoplasmic pathology and delayed apoptotic changes. Ad -CTS1 induces prominent activation of various p53 target genes, including p 21 and MDM-2, but has no relevant effects on BCL-2 family protein expressio n. Although Ad-CTS1 strongly enhances CD95 expression at the cell surface, endogenous CD95/CD95 ligand interactions do not mediate CTS1-induced cell d eath. This is because Ad-CTS1 promotes neither caspase activation nor mitoc hondrial cytochrome c release and because the caspase inhibitors, z-val-Ala -DL-Asp-fluoroniethylketone (zVAD)-fmk or z-Ile-Glu-Thr-Asp-fluoromethylket one (z-IETD)-fmk, do not block CTS1-induced cell death. Ad-CTS1 synergizes with radiotherapy and CD95 ligand in killing glioma cells. In summary, Ad-C TS1 induces an unusual type of cell death that appears to be independent of BCL-2 family proteins, cytochrome c release, and caspases. CTS1 gene trans fer is a promising strategy of somatic gene therapy for malignant glioma.