Gallo-Penn A M, Shirley P S, Andrews J L, Kayda D B, Pinkstaff A M, Kaloss M, Tinlin S, Cameron C, Notley C, Hough C, Lillicrap D, Kaleko M, Connelly S
Genetic Therapy, Inc., a Novartis Company, Gaithersburg, MD 20878, USA.
Hum Gene Ther. 1999 Jul 20;10(11):1791-802. doi: 10.1089/10430349950017473.
Hemophilia A is the most common severe hereditary coagulation disorder and is caused by a deficiency in blood clotting factor VIII (FVIII). Canine hemophilia A represents an excellent large animal model that closely mimicks the human disease. In previous studies, treatment of hemophiliac dogs with an adenoviral vector encoding human FVIII resulted in complete correction of the coagulation defect and high-level FVIII expression [Connelly et al. (1996). Blood 88, 3846]. However, FVIII expression was short term, limited by a strong antibody response directed against the human protein. Human FVIII is highly immunogenic in dogs, whereas the canine protein is significantly less immunogenic. Therefore, sustained phenotypic correction of canine hemophilia A may require the expression of the canine protein. In this work, we have isolated the canine FVIII cDNA and generated an adenoviral vector encoding canine FVIII. We demonstrate expression of canine FVIII in hemophiliac mice at levels 10-fold higher than those of the human protein expressed from an analogous vector. Canine FVIII expression was sustained above human therapeutic levels (50 mU/ml) for at least 1 year in hemophiliac mice.
甲型血友病是最常见的严重遗传性凝血障碍疾病，由血液凝血因子VIII（FVIII）缺乏引起。犬类甲型血友病是一种极佳的大型动物模型，能非常接近地模拟人类疾病。在先前的研究中，用编码人FVIII的腺病毒载体治疗血友病犬，可使凝血缺陷得到完全纠正，并实现高水平的FVIII表达[康奈利等人（1996年）。《血液》88卷，第3846页]。然而，FVIII的表达是短期的，受到针对人蛋白的强烈抗体反应的限制。人FVIII在犬类中具有高度免疫原性，而犬类蛋白的免疫原性则显著较低。因此，要持续表型纠正犬类甲型血友病可能需要表达犬类蛋白。在这项研究中，我们分离出了犬类FVIII cDNA，并构建了一个编码犬类FVIII的腺病毒载体。我们证明，在血友病小鼠中，犬类FVIII的表达水平比从类似载体表达的人蛋白高10倍。在血友病小鼠中，犬类FVIII的表达持续高于人类治疗水平（50 mU/ml）至少1年。
