Ye X, Robinson M B, Batshaw M L, Furth E E, Smith I, Wilson J M
Institute for Human Gene Therapy, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
J Biol Chem. 1996 Feb 16;271(7):3639-46. doi: 10.1074/jbc.271.7.3639.
A murine model of ornithine transcarbamylase (OTC) deficiency was used in this study to evaluate the efficacy of recombinant adenoviruses for correcting the metabolic defect in liver. Recombinant adenoviruses deleted in E1 and containing a human OTC cDNA expressed little functional OTC enzyme in vivo and had no observable impact on the underlying metabolic abnormalities of the OTC-deficient mouse (i.e. elevated urinary orotate and serum glutamine). E1-deleted vectors were improved through the use of the strong constitutive promoter from cytomegalovirus driving the normal murine homolog of OTC cDNA and the ablation of E2a with a temperature-sensitive mutation. Infusion of this improved vector into the mouse model was associated with a complete normalization of liver OTC enzyme activity that persisted for at least 2 months with complete but transient correction in serum glutamine and urine orotic acid. These studies illustrate the utility of improved adenoviral vectors in the treatment of liver metabolic disease.
本研究使用鸟氨酸转氨甲酰酶(OTC)缺乏的小鼠模型来评估重组腺病毒纠正肝脏代谢缺陷的功效。缺失E1并包含人OTC cDNA的重组腺病毒在体内表达很少的功能性OTC酶,并且对OTC缺陷小鼠的潜在代谢异常(即尿乳清酸升高和血清谷氨酰胺升高)没有可观察到的影响。通过使用来自巨细胞病毒的强组成型启动子驱动OTC cDNA的正常小鼠同源物以及通过温度敏感突变消除E2a来改进缺失E1的载体。将这种改进的载体注入小鼠模型与肝脏OTC酶活性完全正常化相关,这种正常化持续至少2个月,血清谷氨酰胺和尿乳清酸得到完全但短暂的纠正。这些研究说明了改进的腺病毒载体在治疗肝脏代谢疾病中的效用。