RRC ID 12036
Author Maeda M, Namikawa K, Kobayashi I, Ohba N, Takahara Y, Kadono C, Tanaka A, Kiyama H.
Title Targeted gene therapy toward astrocytoma using a Cre/loxP-based adenovirus system.
Journal Brain Res
Abstract The aim of this study was to establish a novel adenovirus-based gene therapy system targeting astrocytoma. For this purpose, the Cre recombinase (Cre)/loxP system together with the astrocytoma-specific promoter for GFAP were used. We constructed an adenovirus (Ad) vector that expressed Cre under the control of the GFAP promoter (AxGFAPNCre), as well as another Ad vector containing a switching unit. The latter vector contained a stuffer sequence encoding GFP (AxCALGLTK) with a functional polyadenylation signal between two loxP sites, followed by the herpes simplex virus thymidine kinase (HSV-TK) gene under the control of the CAG promoter. In this system, gene expression of either the stuffer sequence (GFP) or the downstream gene (HSV-TK) was switched on by co-expression of Cre recombinase. Western blot analysis demonstrated specific expression of high levels of TK protein in C6 glioma cells after co-infection of AxGFAPNCre and AxCALGLTK. In vivo, AxGFAPNCre/AxCALGLTK injection into C6 gliomas in the subcutaneous tissue of nude mice followed by intraperitoneal ganciclovir (GCV) treatment significantly suppressed tumor growth compared with control mice. Co-infection of AxGFAPNCre and AxCALNLLacZ resulted in LacZ expression in C6 glioma cells and some reactive astrocytes, whereas GFP was expressed in other cell types surrounding the injected site. Furthermore, a combination of AxGFAPNCre/AxCALGLTK and intraperitoneal GCV injection significantly regressed intracranial C6 gliomas in the rat striatum and prolonged the survival time compared with control rats. The present results indicate that this cell-type-specific gene therapy using a Cre/loxP adenovirus system is both operational and effective, at least against astrocytoma.
Volume 1081(1)
Pages 34-43
Published 2006-4-7
DOI 10.1016/j.brainres.2006.01.105
PII S0006-8993(06)00252-6
PMID 16529724
MeSH Adenoviridae / genetics Animals Astrocytoma / pathology Astrocytoma / therapy* Blotting, Western / methods Brain Neoplasms / pathology Brain Neoplasms / therapy Brain Neoplasms / virology Dose-Response Relationship, Drug Extracellular Matrix Proteins / biosynthesis Extracellular Matrix Proteins / genetics Extracellular Matrix Proteins / therapeutic use* Gene Transfer Techniques Genetic Therapy* / methods Humans Immunohistochemistry / methods Integrases / biosynthesis Integrases / genetics Integrases / therapeutic use* Male Mice Mice, Inbred BALB C Mice, Nude Neoplasm Transplantation / methods Protein-Lysine 6-Oxidase / biosynthesis Protein-Lysine 6-Oxidase / genetics Protein-Lysine 6-Oxidase / therapeutic use* Rats Rats, Wistar Time Factors Tumor Cells, Cultured Viral Proteins / biosynthesis Viral Proteins / genetics Viral Proteins / therapeutic use*
IF 2.929
Times Cited 15
DNA material pCALwL (RDB1679)