De Bishnu P, Heguy Adriana, Hackett Neil R, Ferris Barbara, Leopold Philip L, Lee John, Pierre Lorraine, Gao Guangping, Wilson James M, Crystal Ronald G
Belfer Gene Therapy Core Facility, Weill Medical College of Cornell University, New York, NY 10021, USA.
Mol Ther. 2006 Jan;13(1):67-76. doi: 10.1016/j.ymthe.2005.09.003. Epub 2005 Nov 2.
Alpha1-antitrypsin (alpha1AT) deficiency is a genetic disorder causing emphysema if serum alpha1AT levels are <570 microg/ml. We have shown that intrapleural administration of an AAV5alpha1AT vector yielded persistent therapeutic alpha1AT serum levels. Since anti-AAV2 and -AAV5 antibodies prevalent in humans may limit the use of these common serotypes in gene therapy, we screened 25 AAV vectors derived from humans and nonhuman primates for alpha1AT expression following intrapleural administration to mice. The rhesus AAVrh.10 serotype yielded the highest levels and was chosen for further study. Following intrapleural administration, 77% of total body transgene expression was in the chest wall, diaphragm, lung, and heart. Intrapleural administration of AAVrh.10alpha1AT provided long-term, therapeutic alpha1AT expression in mice, although higher doses were required to achieve therapeutic levels in female mice than in male mice. Intrapleural administration of AAVrh.10alpha1AT produced the same levels in AAV2/AAV5-preimmune and naive mice. In mice administered with AAV5alpha1AT and subsequently "boosted" with the AAVrh.10alpha1AT vector, serum levels were increased by 300%. These data indicate that AAVrh.10 is the most effective known AAV vector for intrapleural gene delivery and has the advantage of circumventing human immunity to AAV.
α1-抗胰蛋白酶（α1AT）缺乏症是一种遗传性疾病，如果血清α1AT水平<570微克/毫升，会导致肺气肿。我们已经表明，胸膜腔内给予AAV5α1AT载体可产生持续治疗性的α1AT血清水平。由于人类中普遍存在的抗AAV2和抗AAV5抗体可能会限制这些常见血清型在基因治疗中的应用，我们筛选了25种源自人类和非人类灵长类动物的AAV载体，以研究其胸膜腔内给予小鼠后α1AT的表达情况。恒河猴AAVrh.10血清型产生的水平最高，并被选用于进一步研究。胸膜腔内给药后，全身转基因表达的77%存在于胸壁、膈肌、肺和心脏中。胸膜腔内给予AAVrh.10α1AT可在小鼠中提供长期的治疗性α1AT表达，尽管与雄性小鼠相比，雌性小鼠需要更高的剂量才能达到治疗水平。胸膜腔内给予AAVrh.10α1AT在AAV2/AAV5预免疫小鼠和未免疫小鼠中产生的水平相同。在给予AAV5α1AT并随后用AAVrh.10α1AT载体“加强”的小鼠中，血清水平提高了300%。这些数据表明，AAVrh.10是已知的用于胸膜腔内基因递送最有效的AAV载体，并且具有规避人类对AAV免疫的优势。
