Miyanohara A, Yee J K, Bouic K, LaPorte P, Friedmann T
Department of Pediatrics, UCSD School of Medicine, La Jolla, CA 92093.
Gene Ther. 1995 Mar;2(2):138-42.
Ideal methods for human gene therapy will eventually include direct gene transfer to defective tissues in a patient in vivo. Toward that goal, we have used high titer, pseudotyped retroviral vectors expressing genes for the Escherichia coli beta-galactosidase (lacZ) or hepatitis B virus surface antigen (HBsAg) to infect mouse liver by in vivo direct injection into the liver parenchyma. We have found that a single percutaneous injection of small volumes of vectors into the newborn mouse liver leads to transduction of at least 25-30% of the hepatocytes throughout the liver, as judged by in situ staining of liver sections for beta-gal activity at 4 weeks after injection. We have demonstrated that stable levels of HBsAg were also detected in the circulation of injected mice up to 4 months after HBsAg-vector injection. We suggest that the high efficiency of in vivo transduction in the neonatal liver and subsequent stable transgene expression by high-titer pseudotyped retroviral vectors in the absence of an invasive partial hepatectomy may effectively be applied to gene therapy studies in a number of human liver disease [corrected].
人类基因治疗的理想方法最终将包括在患者体内直接将基因转移到有缺陷的组织中。为了实现这一目标，我们使用了表达大肠杆菌β-半乳糖苷酶（lacZ）或乙型肝炎病毒表面抗原（HBsAg）基因的高滴度、假型逆转录病毒载体，通过向肝实质内进行体内直接注射来感染小鼠肝脏。我们发现，在新生小鼠肝脏中单次经皮注射少量载体，通过注射后4周对肝脏切片进行β-半乳糖苷酶活性的原位染色判断，可导致整个肝脏中至少25%-30%的肝细胞发生转导。我们还证明，在注射HBsAg载体后长达4个月的时间里，在注射小鼠的循环系统中也能检测到稳定水平的HBsAg。我们认为，在新生肝脏中体内转导的高效率以及随后在无侵入性部分肝切除术的情况下高滴度假型逆转录病毒载体实现的稳定转基因表达，可能有效地应用于多种人类肝脏疾病的基因治疗研究[已修正]。
