論文 - 詳細
| RRC ID | 35679 |
|---|---|
| 著者 | Yoshida Y, Sadata A, Zhang W, Saito K, Shinoura N, Hamada H. |
| タイトル | Generation of fiber-mutant recombinant adenoviruses for gene therapy of malignant glioma. |
| ジャーナル | Hum Gene Ther |
| Abstract |
Recombinant adenovirus (Adv)-mediated gene transduction is a powerful technology for cancer gene therapy. In this article, we report the generation of a fiber-mutant Adv vector, using the Adv genomic DNA-terminal protein complex (DNA-TPC) cotransfection method. First, a fiber-mutant construct in a plasmid carrying the right-side two-thirds of the human adenovirus type 5 (Ad5) genome (pTR) was cotransfected with Ad5 DNA-TPC, yielding the recombinant Adv with the desired fiber mutation. The DNA-TPC from the mutant Adv was then utilized to produce a second-step recombinant Adv with an expression cassette in the place of E1. By this procedure, we generated a fiber mutant, F/K20, that has a linker and a stretch of 20 lysine residues added at the C terminus of the fiber. By using Adv carrying a reporter lacZ gene (AxCAZ2) with either F/K20 or wild-type fiber (F/wt), we examined the transduction efficiency of F/K20-Adv. No significant difference in the transduction efficiency between F/K20 and F/wt-Adv was observed for a human fibroblast line, WI-38, or various tumor cell lines, including melanoma, prostate, esophageal, and pancreatic cancer lines. In clear contrast, F/K20-Adv showed a remarkably enhanced efficiency in genetic transduction of human glioma cells. In all four human glioma lines tested, the multiplicities of infection (MOIs) for transduction of 50% of the population (ED50) were decreased with F/K20-Adv compared with F/wt-Adv: 7-fold for T98G, 14-fold for U251, 9-fold for U373, and 42-fold for U87 cells. Therefore, we attempted to apply F/K20-Adv for gene therapy of malignant glioma. Glioma cells infected with F/K20-Adv carrying genes for interleukin 2 or interleukin 12 produced a high level of each cytokine at a much lower MOI than did cells infected with F/wt-Adv. Infection with F/K20-Adv carrying the wild-type p53 tumor suppressor gene resulted in an enhanced level of p53 protein expression and an increased incidence of F/K20-Adv in transduction efficiency for malignant glioma, providing promising tools for gene therapy. |
| 巻・号 | 9(17) |
| ページ | 2503-15 |
| 公開日 | 1998-11-20 |
| DOI | 10.1089/hum.1998.9.17-2503 |
| PMID | 9853517 |
| MeSH | Adenoviridae / genetics* Apoptosis / genetics Base Sequence Brain Neoplasms / genetics Brain Neoplasms / pathology Brain Neoplasms / therapy* Cell Line DNA Primers Genetic Therapy* Glioma / genetics Glioma / pathology Glioma / therapy* Humans Interleukin-12 / genetics Interleukin-2 / genetics Microscopy, Electron Plasmids Recombination, Genetic Transduction, Genetic Tumor Cells, Cultured |
| IF | 4.51 |
| 引用数 | 82 |
| WOS 分野 | BIOTECHNOLOGY & APPLIED MICROBIOLOGY MEDICINE, RESEARCH & EXPERIMENTAL GENETICS & HEREDITY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
|
| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 15 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 遺伝子材料 | pCAZ 2 (RDB01870) |
| ヒト・動物細胞 | WI-38 |