Left ventricular targeting of reporter gene expression in vivo by human BNP promoter in an adenoviral vector.
To selectively introduce genes into the mouse myocardium, we used a recombinant adenovirus encoding a transgene composed of a cardiac-specific promoter [the proximal human brain natriuretic peptide (hBNP) promoter] coupled to a luciferase reporter gene (Ad.hBNPLuc). Activity in vitro and in vivo was compared with Ad.CMVLuc, which contained the cytomegalovirus (CMV) enhancer/promoter. We tested cell-specific and inducible regulation of Ad.hBNPLuc in vitro. Expression was higher in neonatal cardiac myocytes than in a fibroblast cell line and was induced by interleukin-1beta, phenylephrine, and isoproterenol in myocytes. For in vivo experiments, Ad.hBNPLuc, Ad.CMVLuc, or vehicle was injected into the left ventricular (LV) free wall of the mouse heart. In Ad.hBNPLuc-injected mice, luciferase activity was only detected in the heart. In contrast, Ad.CMVLuc-injected mice had detectable luciferase activity in all tissues examined. Our studies indicate that 1) the cardiac-specific hBNP promoter and direct cardiac injection limit expression of the transgene to the LV free wall; and 2) transgene expression in vitro is inducible and cardiac myocyte specific. Thus the use of the proximal hBNP promoter in recombinant adenoviral vectors may allow cardiac-specific and inducible expression of therapeutic genes in vivo and prevent some of the side effects of systemic adenovirus administration.
LaPointe
Margot C
MC
Hypertension and Vascular Research Division, Henry Ford Hospital, Detroit, Michigan 48202, USA. mclapointe@aol.com
Yang
Xiao-Ping
XP
Carretero
Oscar A
OA
He
Quan
Q
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology; D015870:Gene Expression / Q000502:physiology; D017930:Genes, Reporter; D005822:Genetic Vectors; D006321:Heart / Q000502:physiology*; D006801:Humans; D008156:Luciferases / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008845:Microinjections; D018485:Muscle Fibers, Skeletal / Q000502:physiology; D009206:Myocardium / Q000166:cytology; D020097:Natriuretic Peptide, Brain / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D016276:Ventricular Function
D020097:Natriuretic Peptide, Brain; D008156:Luciferases