Widespread long-term gene transfer to mouse skeletal muscles and heart.

Abstract

Methods

Angiogenesis induced in muscle by a recombinant adenovirus expressing functional isoforms of basic fibroblast growth factor

“…10,11,13 An improvement in myocardial blood flow was reported after intracoronary injection of a recombinant adenovirus expressing FGF5 in the pig ischemic heart. 16 Recently, the recombinant Ad expressing VEGF 121 was shown to protect against ischemic vascular occlusion in a rat limb and to improve myocardial perfusion in the porcine ischemic heart. 14,15 In the study at hand, the human bFGF gene was expressed under the control of the RSV promoter.…”

“…16 The E1-defective virus Ad-RSVbFGF, expressing the bFGF cDNA under the control of the promoter of the Rous sarcoma virus (RSV) was constructed as follows. The plasmid pAd-RSVbFGF was obtained after subcloning of a XhoI/EcoRV fragment (1.62 kb) from the pSCT40 expression vector containing the sequence from nucleotide 295 to 1919 of the human bFGF cDNA isolated by Prats et al, 18 into the plasmid pAd-LTRIX cleaved by SalI and EcoRV.…”

“…[10][11][12][13][14][15] Ad vectors present several advantages including: (1) a low pathogenicity in nonimmunosuppressed individuals; (2) a high capacity to accept foreign DNA; (3) high viral titers; (4) an efficient gene expression in quiescent cells such as skeletal muscle fibers or cardiomyocytes and (5) a transient expression in dividing cells, which is particularly adapted to the time course of vessel neoformation. 16 In counterpart, adenovirus vectors induce inflammatory and immune responses and the current vectors employed are not tissue specific. 17 In the present study we have demonstrated that the bFGF isoforms can be expressed in a fully functional manner in cultured myoblasts or muscle fibres infected with a replication-deficient adenovirus bearing the human bFGF (Ad-RSVbFGF).…”

Imaging and tissue biodistribution of 99mTc-labeled adenovirus knob (serotype 5)

“…Among viral vectors, adenoviruses possess the unique attribute of stability in the bloodstream, a feature which should permit development of the adenovirus as a gene therapy vector which can be administered to the patient simply by intravenous injection. 1,2 However, intravenous administration of the present generation of Ad vectors delivers more than 90% of the input virus to the liver, thereby reducing the titer of virus particles available for transduction of the target disease cells. [2][3][4] Therefore, it will be necessary to overcome this problem of hepatic sequestration if the adenovirus is to realize its potential as an effective vector for gene therapy.…”

Adenovirus-mediated gene transfer in dog prostate: a preclinical study of a relevant model system for gene therapy of human prostatic cancer

“…13 Viral stocks were prepared on 293 cells, as previously described, 14,15 and puri®ed twice on isopicnic CsCl gradiant. Desalting was performed using Pharmacia columns (Orsay, France).…”