Lipshutz G S, Gruber C A, Hardy J, Contag C H, Gaensler K M
Department of Surgery, University of California, San Francisco, San Francisco, California 94143-0793, USA.
Mol Ther. 2001 Mar;3(3):284-92. doi: 10.1006/mthe.2001.0267.
Recombinant adeno-associated viruses (rAAV) are promising gene transfer vectors that produce long-term expression without toxicity. To investigate future approaches for in utero gene delivery, the efficacy and safety of prenatal administration of rAAV were determined. Using luciferase as a reporter, expression was assessed by whole-body imaging and by analysis of luciferase activity in tissue extracts, at the time of birth and monthly thereafter. Transgene expression was detected in all injected animals. Highest levels of luciferase activity were detected at birth in the peritoneum and liver, while the heart, brain, and lung demonstrated low-level expression. In vivo luciferase imaging revealed persistent peritoneal expression for 18 months after in utero injection and provided a sensitive whole-body assay, useful in identifying tissues for subsequent analyses. There was no detectable hepatocellular injury. Antibodies that reacted with either luciferase or rAAV were not found. AAV sequences were not detected in germ-line tissues of injected animals or in tissues of their progeny. In utero AAV-mediated gene transfer in this animal model demonstrates that novel therapeutic vectors and strategies can be rapidly tested in vivo and that rAAV may be developed to ameliorate genetic diseases with perinatal morbidity and mortality.
重组腺相关病毒（rAAV）是很有前景的基因转移载体，可产生长期表达且无毒性。为了研究子宫内基因递送的未来方法，我们确定了产前给予rAAV的疗效和安全性。以荧光素酶作为报告基因，在出生时及之后每月通过全身成像和分析组织提取物中的荧光素酶活性来评估表达情况。在所有注射的动物中均检测到转基因表达。出生时在腹膜和肝脏中检测到最高水平的荧光素酶活性，而心脏、大脑和肺则表现出低水平表达。体内荧光素酶成像显示子宫内注射后腹膜持续表达18个月，并提供了一种灵敏的全身检测方法，有助于识别后续分析的组织。未检测到肝细胞损伤。未发现与荧光素酶或rAAV反应的抗体。在注射动物的生殖系组织或其后代的组织中未检测到AAV序列。在该动物模型中，子宫内AAV介导的基因转移表明，新型治疗载体和策略可在体内快速进行测试，并且rAAV可用于改善具有围产期发病率和死亡率的遗传性疾病。
