TAKEUCHI Toshiyuki Gunma University, Institution of molecular regulation, Professor, 生態調節研究所, 教授 (00109977)
小林 聡 群馬大学, 医学部, 助手 (00265779)
石内 勝吾 群馬大学, 医学部, 助手 (10312878)
|Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2002: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 2001: ¥2,000,000 (Direct Cost: ¥2,000,000)
Glioma is one of the most common primary brain tumors, and develops invasively in the normal brain tissues. Therefore, in the gene therapy of glioma using sulcide gene, such as p53, glioma-specific gene expression is a prerequisite for prevention against the normal brain injury. We have developed a glioma-specific adenoviral gene expression vector using a "nestin" regulatory element. Nestin is a central nervous system (CNS) progenitor cell-specific intermediate filament, and the incidence of nestin expression correlates well with malignancy of glioma. We have constructed glioma-specific "regulator" recombinant adenovirus vector, Ax2iNPNCre, which contains the site-specific recombinase Cre under the control of the rat nestin regulatory element 2iNP. For the treatment of glioma, we have constructed AxCALNLNp53K, which activate the p53 gene under the control of a potent CAG promoter by the Cre-producing adenoviruses. For assessing the cell toxicity of this adenoviral system, we used two p53 mutated human glioma cell line, U251, which highly express nestin gene, and T98G, which little express nestin gene. In vitro co-transfection of Ax2iNPNCre and AxCALNLNp53K induced the cell death to U251 within three days after the infection, but not to T98G. Co-transfection of AxCANCre (control vector which highly express Cre in almost eucaryotic cells, and AxCALNLNp53K induced the cell death to T98G. In the animal model of glioma by implanting U251 human glioma cells into subcutaneous tissues, this adenovirus vector system augmented the tumor growth. Thus, nestin positive cell selective induction of cell death was achieved by this double infection therapy.