Wang Alfred Y, Peng Peter D, Ehrhardt Anja, Storm Theresa A, Kay Mark A
Department of Molecular Pharmacology, School of Medicine, Stanford University, Stanford, CA 94305, USA.
Hum Gene Ther. 2004 Apr;15(4):405-13. doi: 10.1089/104303404322959551.
Although effective gene therapy vectors have been developed for organ systems such as the liver, an effective delivery vector to the pancreas in vivo has remained elusive. Of the currently available viral vectors, adenovirus and adeno-associated virus (AAV) are two of the most efficient at transducing nondividing cells. We have constructed recombinant adenovirus (AdVLacZ), adeno-associated virus serotype 2 (AAV2LacZ), and pseudotyped adeno-associated virus serotype 5 and 8 (AAV5LacZ, AAV8LacZ) carrying the LacZ reporter, and compared the transduction efficiency of these four vectors in the pancreas of mice in vivo. We showed that adenovirus, AAV2, and AAV8 are capable of transducing the pancreas in vivo, but with different expression kinetics, efficiencies of transduction, and persistence. AdVLacZ-transduced pancreas exhibited maximum LacZ expression at 1 week postdelivery, with greater than 90% of expression lost at 4 weeks. AAV2LacZ-transduced pancreas displayed peak LacZ levels at 4 weeks postdelivery, with no significant decrease in expression for up to 8 weeks. AAV8LacZ was at least 10-fold more efficient than AAV2LacZ in transducing the pancreas in vivo, with significant levels of expression detectable at 1 week, whereas AAV5LacZ did not result in any detectable transgene expression at all tested time points. All three vectors primarily transduced pancreatic acinar cell types, with limited transduction of pancreatic endocrine cells. AdVLacZ elicited a significant leukocyte infiltration early after delivery into the pancreas, whereas none of the AAV vectors elicited a significant leukocyte response. None of the tested vectors caused significant changes in serum amylase or blood glucose levels, suggesting that they do not significantly alter pancreatic function. These vectors will be useful for studying novel gene delivery based treatments in animal models for diabetes and other pancreatic disorders.
尽管已经为肝脏等器官系统开发出了有效的基因治疗载体，但一种能够在体内有效递送至胰腺的载体仍难以找到。在目前可用的病毒载体中，腺病毒和腺相关病毒（AAV）是转导非分裂细胞最有效的两种载体。我们构建了携带LacZ报告基因的重组腺病毒（AdVLacZ）、2型腺相关病毒（AAV2LacZ）以及5型和8型假型腺相关病毒（AAV5LacZ、AAV8LacZ），并比较了这四种载体在小鼠胰腺中的体内转导效率。我们发现腺病毒、AAV2和AAV8能够在体内转导胰腺，但它们的表达动力学、转导效率和持久性各不相同。AdVLacZ转导的胰腺在递送后1周时LacZ表达达到最大值，4周时超过90%的表达消失。AAV2LacZ转导的胰腺在递送后4周时LacZ水平达到峰值，长达8周表达无显著下降。AAV8LacZ在体内转导胰腺的效率比AAV2LacZ至少高10倍，在1周时即可检测到显著水平的表达，而AAV5LacZ在所有测试时间点均未导致任何可检测到的转基因表达。所有三种载体主要转导胰腺腺泡细胞类型，对胰腺内分泌细胞的转导有限。AdVLacZ在递送至胰腺后早期引发了显著的白细胞浸润，而AAV载体均未引发显著的白细胞反应。所有测试载体均未导致血清淀粉酶或血糖水平发生显著变化，表明它们不会显著改变胰腺功能。这些载体将有助于在糖尿病和其他胰腺疾病的动物模型中研究基于新型基因递送的治疗方法。
