621. Angiogenesis and Arteriogenesis in Ischemic Skeletal Muscles Following VEGF Gene Transfer by Adeno-Associated Virus Serotype 1 Vectors
| Autor: | Ling-yun Zu, Hua Yan, Xiaobing Wu, Wei Gao, Peng Zhang, Yanhong Guo |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
Pharmacology
Gene isoform Reporter gene Myogenesis Angiogenesis Biology medicine.disease_cause Molecular biology Vascular endothelial growth factor chemistry.chemical_compound chemistry Drug Discovery Gene expression Genetics medicine Molecular Medicine Vector (molecular biology) Molecular Biology Adeno-associated virus |
| Zdroj: | Molecular Therapy. 11:S241 |
| ISSN: | 1525-0016 |
| Popis: | Top of pageAbstract Recombinant adeno-associated viruses (rAAVs) have been widely explored for gene transfer into skeletal muscles. Serotype 2 rAAV (rAAV2) vectors have been most frequently employed. rAAV2 based vectors peudotyped with serotype 1 capsid proteins (rAAV1 vectors) may allow superior gene transfer into skeletal muscles. To assess the potential utility of rAAV1 vector gene transfer in ischemic skeletal muscles, we have generated rAAV1 vectors encoding green fluorescence protein (GFP) or LacZ as reporter gene or the 165 aa isoform of human vascular endothelial growth factor (VEGF165) as functional gene. In C2C12 derived myotubes the rAAV1-GFP vector allowed 3- to 4-fold higher gene expression in comparison with the rAAV2-GFP vector. VEGF secretion from C2C12 derived myotubes infected with rAAV1-VEGF165 vector increased significantly compared with that of rAAV2-165 vector. Following establishing ischemia in mouse hind limbs, high titer rAAV1 or rAAV2 vector preparations encoding LacZ were injected in ischemic thigh muscles. We demonstrated that the rAAV1-LacZ vector allowed higher level, broader distribution and longer lasting gene expression compared with the rAAV2-LacZ vector. At 6 months post rAAV vector injection substantial fraction of the muscle fibers were still expressing |[beta]|-gal in rAAV1-LacZ vector injected ischemic muscles, in contrast, |[beta]|-gal expression was barely detectable in rAAV2-LacZ vector injected ischemic muscles. Muscle VEGF165 production following the rAAV1-VEGF165 vector injection was 5 to 10 times higher than that following the rAAV2-VEGF165 vector injection. The average VEGF concentration was 205 |[plusmn]| 88 pg/mg total protein in the rAAV1-VEGF165 vector injected muscles compared with 38 |[plusmn]|22 pg/mg total protein in the rAAV2-VEGF165 injected muscles (P < 0.001, Fig. 1). High level VEGF165 production following the rAAV1-VEGF165 vector injection stimulated superior muscle regeneration, angiogenesis and arteriogenesis with the formation of relatively mature vascular structures in the previously ischemic skeletal muscles (Fig. 2). Thus, the rAAV1-VEGF165 vector mediated gene transfer can be a powerful tool for muscle ischemia treatment. |
| Databáze: | OpenAIRE |
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