Preclinical Evaluation of a Recombinant Adeno-Associated Virus Vector Expressing Human Alpha-1 Antitrypsin Made Using a Recombinant Herpes Simplex Virus Production Method
Preclinical Evaluation of a Recombinant Adeno-Associated Virus Vector Expressing Human Alpha-1 Antitrypsin Made Using a Recombinant Herpes Simplex Virus Production Method
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DOI：
10.1089/hum.2010.118
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发表时间：
2011-02-01
影响因子：
4.2
通讯作者：
Flotte, Terence R.
中科院分区：
文献类型：
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作者：
Chulay, Jeffrey D.;Ye, Guo-Jie;Flotte, Terence R.
重组腺相关病毒（rAAV）载体为α -1抗胰蛋白酶（AAT）缺乏症的基因治疗提供了希望。一项小鼠毒理学研究评估了用单纯疱疹病毒（HSV）互补系统或质粒转染（TFX）方法肌肉注射表达人AAT （rAAV- cb - haat）的rAAV载体，两种载体的剂量均为3 × 10(11) vg (1.2 × 10(13) vg/kg)，单纯疱疹病毒产生的载体的剂量为2 × 10(12) vg （8 × 10(13) vg/kg）。该单纯疱疹病毒载体在纯度、转导效率和hAAT表达方面具有良好的体外特性。试验品与对照组的临床表现、血液学和临床化学值均无显著差异，病理检查无明显差异。组织病理学检查显示注射部位骨骼肌有轻微到轻微的变化，包括局灶性慢性间质炎症和肌肉变性、再生和空泡化。在3x10(11) vg剂量下，hsv产生的载体的血清hAAT水平高于tfx产生的载体。随着单纯疱疹病毒载体剂量的增加，血清hAAT水平的增加在女性中呈剂量正比，在男性中大于剂量正比。在给药后24小时，血液中的载体拷贝数最高，此后下降，在给药后90天没有检测到拷贝。在几乎所有载体处理的动物中检测到hAAT抗体，在接受最高载体剂量的大多数动物中检测到HSV抗体。这些结果支持继续开发rAAV-CB-hAAT治疗AAT缺乏症。
Recombinant adeno-associated virus (rAAV) vectors offer promise for gene therapy of alpha-1 antitrypsin (AAT) deficiency. A toxicology study in mice evaluated intramuscular injection of an rAAV vector expressing human AAT (rAAV-CB-hAAT) produced using a herpes simplex virus (HSV) complementation system or a plasmid transfection (TFX) method at doses of 3x10(11) vg (1.2x10(13) vg/kg) for both vectors and 2x10(12) vg (8x10(13) vg/kg) for the HSV-produced vector. The HSV-produced vector had favorable in vitro characteristics in terms of purity, efficiency of transduction, and hAAT expression. There were no significant differences in clinical findings or hematology and clinical chemistry values between test article and control groups and no gross pathology findings. Histopathological examination demonstrated minimal to mild changes in skeletal muscle at the injection site, consisting of focal chronic interstitial inflammation and muscle degeneration, regeneration, and vacuolization, in vector-injected animals. At the 3x10(11) vg dose, serum hAAT levels were higher with the HSV-produced vector than with the TFX-produced vector. With the higher dose of HSV-produced vector, the increase in serum hAAT levels was dose-proportional in females and greater than dose-proportional in males. Vector copy numbers in blood were highest 24 hr after dosing and declined thereafter, with no detectable copies present 90 days after dosing. Antibodies to hAAT were detected in almost all vector-treated animals, and antibodies to HSV were detected in most animals that received the highest vector dose. These results support continued development of rAAV-CB-hAAT for treatment of AAT deficiency.