Efficient and Targeted Transduction of Nonhuman Primate Liver With Systemically Delivered Optimized AAV3B Vectors
Efficient and Targeted Transduction of Nonhuman Primate Liver With Systemically Delivered Optimized AAV3B Vectors
复制标题
DOI：
10.1038/mt.2015.174
复制
发表时间：
2015-12-01
影响因子：
12.4
通讯作者：
Gao, Guangping
中科院分区：
文献类型：
--
作者：
Li, Shaoyong;Ling, Chen;Gao, Guangping
重组腺相关病毒血清3B型(RAAV3B)能有效转导培养的人肝癌细胞和原代人肝细胞。丝氨酸(S)和苏氨酸(T)诱导的衣壳修饰进一步增强了其转导效率。系统递送的衣壳优化的rAAV3B载体可以在人肝癌异种移植模型中特异性地靶向癌细胞，这表明它们可能用于人类肝脏导向的基因治疗。在这里，我们比较了AAV3B和AAV8载体在原代培养的人肝细胞和癌细胞中的转导效率，以及在人肝移植NSG-PIZ小鼠模型中的人和小鼠肝细胞的转导效率。我们还检测了野生型(WT)和衣壳优化型rAAV3B在非人灵长类动物(NHP)肝脏中的安全性和转导效率。静脉递送S663V+T492V(ST)修饰的自互补(Sc)AAV3B-EGFP载体可导致肝脏靶向性增强的绿色荧光蛋白(EGFP)在NHP中的表达，而没有明显的肝脏毒性。静脉注射WT和ST修饰的rAAV3B。重组人绒毛膜促性腺激素(HCG)表达载体在NHP中也能稳定表达绒毛膜促性腺激素。载体基因组主要针对肝脏。临床化学和组织病理学检查显示没有明显的媒介相关毒性。我们的研究对于临床开发优化的AAV3B载体用于人类肝脏导向的基因治疗具有重要意义和参考价值。
Recombinant adeno-associated virus serotype 3B (rAAV3B) can transduce cultured human liver cancer cells and primary human hepatocytes efficiently. Serine (S)- and threonine (T)-directed capsid modifications further augment its transduction efficiency. Systemically delivered capsid-optimized rAAV3B vectors can specifically target cancer cells in a human liver cancer xenograft model, suggesting their potential use for human liver-directed gene therapy. Here, we compared transduction efficiencies of AAV3B and AAV8 vectors in cultured primary human hepatocytes and cancer cells as well as in human and mouse hepatocytes in a human liver xenograft NSG-PiZ mouse model. We also examined the safety and transduction efficacy of wild-type (WT) and capsid-optimized rAAV3B in the livers of nonhuman primates (NHPs). Intravenously delivered S663V+T492V (ST)-modified self-complementary (sc) AAV3B-EGFP vectors led to liver-targeted robust enhanced green fluorescence protein (EGFP) expression in NHPs without apparent hepatotoxicity. Intravenous injections of both WT and ST-modified rAAV3B. ST-rhCG vectors also generated stable super-physiological levels of rhesus chorionic gonadotropin (rhCG) in NHPs. The vector genome predominantly targeted the liver. Clinical chemistry and histopathology examinations showed no apparent vector- related toxicity. Our studies should be important and informative for clinical development of optimized AAV3B vectors for human liver-directed gene therapy.