Gau Chia-Ling, Rosenblatt Robin A, Cerullo Vincenzo, Lay Fides D, Dow Adrienne C, Livesay Justin, Brunetti-Pierri Nicola, Lee Brendan, Cederbaum Stephen D, Grody Wayne W, Lipshutz Gerald S
Department of Human Genetics, David Geffen School of Medicine at UCLA, Los Angeles, California, USA.
Mol Ther. 2009 Jul;17(7):1155-63. doi: 10.1038/mt.2009.65. Epub 2009 Apr 14.
Neonatal gene therapy has the potential to ameliorate abnormalities before disease onset. Our gene knockout of arginase I (AI) deficiency is characterized by increasing hyperammonemia, neurological deterioration, and early death. We constructed a helper-dependent adenoviral vector (HDV) carrying AI and examined for correction of this defect. Neonates were administered 5 x 10(9) viral particles/g and analyzed for survival, arginase activity, and ammonia and amino acids levels. The life expectancy of arg(-/-) mice increased to 27 days while controls died at 14 days with hyperammonemia and in extremis. Death correlated with a decrease in viral DNA/RNA per cell as liver mass increased. Arginase assays demonstrated that vector-injected hepatocytes had ~20% activity of heterozygotes at 2 weeks of age. Hepatic arginine and ornithine in treated mice were similar to those of saline-injected heterozygotes at 2 weeks, whereas ammonia was normal. By 26 days, arginase activity in the treated arg(-/-) livers declined to <10%, and arginine and ornithine increased. Ammonia levels began increasing by day 25, suggesting the cause of death to be similar to that of uninjected arg(-/-) mice, albeit at a later time. These studies demonstrate that the AI deficient newborn mouse can be temporarily corrected and rescued using a HDV.
新生儿基因治疗有潜力在疾病发作前改善异常情况。我们对精氨酸酶I(AI)缺乏症进行的基因敲除表现为高氨血症增加、神经功能恶化和早期死亡。我们构建了携带AI的辅助依赖型腺病毒载体(HDV),并检测其对这种缺陷的纠正情况。给新生儿注射5×10⁹病毒颗粒/克,然后分析其存活率、精氨酸酶活性以及氨和氨基酸水平。arg(-/-)小鼠的预期寿命延长至27天,而对照组在14天时因高氨血症处于濒死状态死亡。随着肝脏质量增加,死亡与每个细胞中病毒DNA/RNA的减少相关。精氨酸酶检测表明,在2周龄时,注射载体的肝细胞具有杂合子约20%的活性。在2周时,治疗小鼠肝脏中的精氨酸和鸟氨酸与注射生理盐水的杂合子相似,而氨水平正常。到26天时,治疗的arg(-/-)肝脏中的精氨酸酶活性降至<10%,精氨酸和鸟氨酸增加。氨水平在第25天开始升高,这表明死亡原因与未注射的arg(-/-)小鼠相似,尽管时间较晚。这些研究表明,使用HDV可以暂时纠正和挽救AI缺乏的新生小鼠。