Intravenous angiotensinogen antisense in AAV-based vector decreases hypertension.
Angiotensinogen (AGT) has been linked to hypertension. Because there are no direct inhibitors of AGT, we have developed antisense (AS) inhibition of AGT mRNA delivered in an adeno-associated virus (AAV)-based plasmid vector. This plasmid, driven by the cytomegalovirus promoter, contains a green fluorescent protein reporter gene and AS cDNA for rat AGT. Transfection of the plasmid into rat hepatoma cells brought a strong expression of the transgenes and a significant reduction in the level of AGT. In the in vivo study, naked plasmid DNA was intravenously injected into adult spontaneously hypertensive rats at different doses (0.6, 1.5, and 3 mg/kg). Expression of AGT AS mRNA was present in liver and heart, and it lasted longer in the liver. All three doses produced a significant decrease in blood pressure (BP). BP decreased for 2, 4, and 6 days, respectively. The lowest dose decreased BP by 12 +/- 3.0 mmHg, whereas the higher doses decreased BP by up to 22.5 +/- 5.2 mmHg compared with the control rats injected with saline (P < 0.01). The injection of the plasmid with liposomes produced a more profound and longer reduction (8 days) in BP. Consistent changes in plasma AGT level were observed. Sense plasmid had no effect. No liver toxicity was observed after injection of AS plasmid with or without liposomes. Our results suggest that the systemic delivery of AS against AGT mRNA by AAV-based plasmid vector, especially with liposomes, may have potential for gene therapy of hypertension and that further studies with the plasmid packaged into a recombinant AAV vector for a longer-lasting AS effect are warranted.
Tang
X
X
Department of Physiology, College of Medicine, University of Florida, Gainesville, Florida 32610, USA.
Mohuczy
D
D
Zhang
Y C
YC
Kimura
B
B
Galli
S M
SM
Phillips
M I
MI
D000808:Angiotensinogen / Q000235:genetics*; D000818:Animals; D001794:Blood Pressure / Q000187:drug effects*; D006528:Carcinoma, Hepatocellular; D003587:Cytomegalovirus / Q000235:genetics; D000229:Dependovirus; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006973:Hypertension / Q000188:drug therapy* / Q000235:genetics* / Q000517:prevention & control; D007275:Injections, Intravenous; D008099:Liver / Q000378:metabolism; D008113:Liver Neoplasms; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D009206:Myocardium / Q000378:metabolism; D020319:Oligodeoxyribonucleotides, Antisense / Q000008:administration & dosage / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D011918:Rats, Inbred SHR; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013599:Systole / Q000187:drug effects; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects*; D014162:Transfection; D014407:Tumor Cells, Cultured
D008164:Luminescent Proteins; D020319:Oligodeoxyribonucleotides, Antisense; D012333:RNA, Messenger; D000808:Angiotensinogen; D049452:Green Fluorescent Proteins