We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
865. In Vivo Gene Transfer of Prostacyclin Synthase by Using AAV Vector Prevents Monocrotaline-Induced Pulmonary Hypertension and Pulmonary Vasoconstriction in Rats.
- Authors
Ito, Takayuki; Okada, Takashi; Mimuro, Jun; Miyashita, Hiroshi; Nonaka-Sarukawa, Mutsuko; Mizukami, Hiroaki; Kume, Akihiro; Takahashi, Masafumi; Yamamoto, Keiji; Shimada, Kazuyuki; Ozawa, Keiya
- Abstract
Background: Prostacyclin synthase (PGIS) is the final committed enzyme for biosynthesis of prostacyclin (PGI2), which ameliorates survival and symptoms in pulmonary arterial hypertension (PAH). However, the therapeutic option of PGI2 is limited by its short half-life, requirement of continuous infusion, and catheter-related complications. Although PGIS gene transfer would effectively and stably express PGI2, previous strategies have several limitations; intratracheal approach may deteriorate respiratory function in critical PAH patients, and intrahepatic approach may cause liver hemorrhage and peritonitis. Thus, intramuscular PGIS gene transfer is considered promising than the others. Nonetheless, a plasmid-based approach has encountered problems arising from transient and insufficient gene expression. Here we employed an adeno-associated virus (AAV) vector for an improved intramuscular gene transfer to prevent monocrotaline (MCT)-induced PAH.Methods: We constructed an AAV type1-based vector containing human PGIS gene driven by the modified chicken β-actin promoter with CMV immediate early enhancer (AAV1-PGIS). Three-week-old Wistar rats were intramuscularly injected with AAV1-PGIS or control vector (AAV1-eGFP) at 1×1010 or 1×1011 g.c./body, followed by MCT (40 mg/kg) injection at 7 weeks. At 11 weeks, we assessed hemodynamic parameters, a weight ratio of right ventricle to left ventricle plus septum {RV/(LV+S)}, and histopathology of pulmonary arteries (PAs). Plasma levels of PGI2 were estimated from the measurement of its stable metabolite, 6-keto prostaglandin F1α (PGF1α), using EIA.Results: Eight weeks after vector injection, the plasma 6-keto PGF1α levels increased in a vector dose-dependent manner (1.62 ± 0.30, untreated; 6.68 ± 1.33, 1×1010 g.c./body; 12.29 ± 3.03 ng/mL, 1×1011 g.c./body). The mean PA pressure, PA resistance, RV/(LV+S), and % medial thickness of PAs in the PGIS group (1×1010 g.c./body) significantly decreased compared to the control group (33.9 ± 2.4 vs. 46.1 ± 3.0 mmHg, p<0.05; 0.26 ± 0.03 vs. 0.41 ± 0.03 mmHg/ mL/min/kg, p<0.05; 0.44 ± 0.03 vs. 0.69 ± 0.01, p<0.01; 14.6 ± 1.5 vs. 23.5 ± 0.5%, p<0.01, respectively). Furthermore, the PGIS-transduced rats exhibited prolonged survivals compared to the eGFP-transduced rats (100% vs. 50% in 8 week, P<0.05, n = 8). Importantly, the PGIS-transduced rats showed no significant complication such as infection and systemic blood pressure reduction.Conclusion: Single intramuscular injection of an AAV vector successfully achieved sustained PGIS expression, leading to PAH prevention. This strategy would provide a promising therapeutic alternative for PAH patients suffering from complications of continuous intravenous PGI2 infusion.Molecular Therapy (2006) 13, S333–S333; doi: 10.1016/j.ymthe.2006.08.953
- Subjects
PROSTACYCLIN; RATS; GENETIC transformation; PERITONITIS; GENE expression; MONOCROTALINE
- Publication
Molecular Therapy, 2006, Vol 13, pS333
- ISSN
1525-0016
- Publication type
Article
- DOI
10.1016/j.ymthe.2006.08.953