AAV serotype-1 mediates early onset of gene expression in mouse hearts and results in better therapeutic effect.
Adeno-associated viral vectors (AAV) are attractive tool for gene therapy for coronary artery disease. However, gene expression in myocardium mediated by AAV serotype 2 (AAV2) does not peak until 4-6 weeks after gene transfer. This delayed gene expression may reduce its therapeutic potential for acute cardiac infarction. To determine whether earlier gene expression and better therapeutic effect could be achieved using a different serotype, CMV promoter driving the EPO gene (AAV-EPO) was packaged into AAV serotypes 1-5 capsids and injected into mouse myocardium. EPO expression was studied by measuring the hematocrits and EPO mRNA. After we found that AAV1 mediates the highest gene expression after 4 days of gene transduction, AAV-LacZ (CMV promoter driving LacZ gene expression) and MLCVEGF (hypoxia-inducible and cardiac-specific VEGF expression) were packaged into AAV1 and 2 capsids. LacZ expression was detected in AAV1-LacZ but not in AAV2-LacZ-injected hearts 1 day after vector injection. Compared to AAV2-MLCVEGF that mediated no significant VEGF expression, AAV1-MLCVEGF mediated 13.7-fold induction of VEGF expression in ischemic hearts 4 days after gene transduction and resulted in more neovasculatures, better cardiac function and less myocardial fibrosis. Thus, AAV1 mediates earlier and higher transgene expression in myocardium and better therapeutic effects.
Su
H
H
Cardiovascular Research Institute, University of California-San Francisco, 513 Parnassus Avenue, San Francisco, CA 94143, USA. hua.su@ucsf.edu
Huang
Y
Y
Takagawa
J
J
Barcena
A
A
Arakawa-Hoyt
J
J
Ye
J
J
Grossman
W
W
Kan
Y W
YW
D000818:Animals; D049109:Cell Proliferation; D003327:Coronary Disease / Q000378:metabolism / Q000628:therapy*; D000229:Dependovirus / Q000235:genetics* / Q000276:immunology; D004452:Echocardiography; D004921:Erythropoietin / Q000235:genetics; D005260:Female; D015870:Gene Expression; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics; D006400:Hematocrit; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D007763:Lac Operon; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D009206:Myocardium / Q000378:metabolism*; D018919:Neovascularization, Physiologic / Q000502:physiology; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012703:Serotyping; D013995:Time; D014161:Transduction, Genetic / Q000379:methods; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism
C497443:Hif1a protein, mouse; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D042461:Vascular Endothelial Growth Factor A; D004921:Erythropoietin