Ito Takayuki, Okada Takashi, Mimuro Jun, Miyashita Hiroshi, Uchibori Ryosuke, Urabe Masashi, Mizukami Hiroaki, Kume Akihiro, Takahashi Masafumi, Ikeda Uichi, Sakata Yoichi, Shimada Kazuyuki, Ozawa Keiya
Division of Genetic Therapeutics, Jichi Medical University, Tochigi, Japan.
Hypertension. 2007 Sep;50(3):531-6. doi: 10.1161/HYPERTENSIONAHA.107.091348. Epub 2007 Jul 16.
Prostacyclin synthase (PGIS) is the final committed enzyme in the metabolic pathway of prostacyclin production. The therapeutic option of intravenous prostacyclin infusion in patients with pulmonary arterial hypertension is limited by the short half-life of the drug and life-threatening catheter-related complications. To develop a better delivery system for prostacyclin, we examined the feasibility of intramuscular injection of an adenoassociated virus (AAV) vector expressing PGIS for preventing monocrotaline-induced pulmonary arterial hypertension in rats. We developed an AAV serotype 1-based vector carrying a human PGIS gene (AAV-PGIS). AAV-PGIS or the control AAV vector expressing enhanced green fluorescent protein was injected into the anterior tibial muscles of 3-week-old male Wistar rats; this was followed by the monocrotaline administration at 7 weeks. Eight weeks after injecting the vector, the plasma levels of 6-keto-prostaglandin F(1alpha) increased in a vector dose-dependent manner. At this time point, the PGIS transduction (1x10(10) genome copies per body) significantly decreased mean pulmonary arterial pressure (33.9+/-2.4 versus 46.1+/-3.0 mm Hg; P<0.05), pulmonary vascular resistance (0.26+/-0.03 versus 0.41+/-0.03 mm Hg x mL(-1) x min(-1) x kg(-1); P<0.05), and medial thickness of the peripheral pulmonary artery (14.6+/-1.5% versus 23.5+/-0.5%; P<0.01) as compared with the controls. Furthermore, the PGIS-transduced rats demonstrated significantly improved survival rates as compared with the controls (100% versus 50%; P<0.05) at 8 weeks postmonocrotaline administration. An intramuscular injection of AAV-PGIS prevents monocrotaline-pulmonary arterial hypertension in rats and provides a new therapeutic alternative for preventing pulmonary arterial hypertension in humans.
前列环素合酶（PGIS）是前列环素生成代谢途径中的最终关键酶。肺动脉高压患者静脉输注前列环素的治疗选择受到药物半衰期短和危及生命的导管相关并发症的限制。为了开发一种更好的前列环素递送系统，我们研究了肌肉注射表达PGIS的腺相关病毒（AAV）载体预防大鼠野百合碱诱导的肺动脉高压的可行性。我们开发了一种携带人PGIS基因的基于AAV血清型1的载体（AAV-PGIS）。将AAV-PGIS或表达增强型绿色荧光蛋白的对照AAV载体注射到3周龄雄性Wistar大鼠的胫前肌中；7周后给予野百合碱。注射载体8周后，血浆6-酮-前列腺素F（1α）水平以载体剂量依赖性方式升高。此时，PGIS转导（每只动物1×10（10）基因组拷贝）与对照组相比，显著降低了平均肺动脉压（33.9±2.4对46.1±3.0 mmHg；P<0.05）、肺血管阻力（0.26±0.03对0.41±0.03 mmHg×mL（-1）×min（-1）×kg（-1）；P<0.05）和外周肺动脉中层厚度（14.6±1.5%对23.5±0.5%；P<0.01）。此外，与对照组相比，PGIS转导的大鼠在给予野百合碱8周后存活率显著提高（100%对50%；P<0.05）。肌肉注射AAV-PGIS可预防大鼠野百合碱诱导的肺动脉高压，并为预防人类肺动脉高压提供了一种新的治疗选择。
