Treatment of glioblastoma by direct inoculation of concentrated high titer-recombinant retrovirus carrying the herpes simplex virus thymidine kinase gene.

摘要: We prepared retroviruses carrying the lacZ gene or herpes simplex virus thymidine kinase (HTK) with titers of 1.4-2.5 x 10(11) colony-forming units (cfu)/ml, and stereotaxically inoculated only 3 microliters into a mouse glioma model. This resulted in highly efficient transduction vivo. The transduced cells migrated far from implantation site, potentiating induction remarkable bystander effect. Following repetitive ganciclovir (GCV) intraperitoneal injection, effective killing brain was observed. efficiency nearly as high that observed for high-titer retrovirus-producing fibroblasts. Eighty per cent tumor-bearing mice were completely cured by our treatment protocol using concentrated HTK-harboring retroviruses. Our results suggest repeated inoculations HTK followed GCV may be promising strategy clinical malignant gliomas. To achieve further safety therapy glioma, genes abundantly expressed human glioblastoma searched Serial Analysis Gene Expression (SAGE) technique. Among top-147 most tags glioblastoma, we found tag, TTTTGGGTAT, originated an unidentified gene, which not detected astrocyte cultures. Real-time quantitative RT-PCR showed MAGE-E1 expression 2.6-15 fold enriched relative to astrocytes. Expressed Sequence Tags (ESTs) containing this tag homologous melanoma-associated antigen (MAGE) family, new cDNA, named MAGE-E1, cloned 5'-rapid amplification cDNA ends (RACE) low other cancers, ovary among normal tissues. These indicate is novel glioma-specific member MAGE can applied transduction.