美国宾夕法尼亚大学James M. Wilson研究组发现，整合的载体基因组可能有助于AAV给药后在灵长类动物肝脏中的长期表达。2023年11月6日，《自然—生物技术》杂志在线发表了这项成果。

研究人员表示，基于肝脏的腺相关病毒（AAV）基因疗法的开发面临着转基因表达效率和持久性有限的问题。

研究人员对静脉注射AAV8和AAVrh10载体超过两年的非人灵长类动物进行了评估，以更好地确定影响性能的转导机制。非免疫原性转基因实现了高度转导，尽管在最初的90天内表达量有所下降，但达到了较低但稳定的状态。超过10%的肝细胞含有载体DNA的单个核域，尽管转基因表达消失了，但这些核域仍然存在。免疫原性转基因的载体DNA和RNA减少幅度更大。每100个细胞中就有1个细胞在广泛分布的位点上检测到载体序列的基因组整合，包括复杂的聚合结构，这些位点并不靠近肝细胞癌相关基因。

这项研究表明，AAV介导的转基因在灵长类肝细胞中的表达分为两个阶段——来自外显子基因组的表达量高但持续时间短，随后是低得多但稳定的表达，很可能来自整合载体。

附：英文原文

Title: Integrated vector genomes may contribute to long-term expression in primate liver after AAV administration

Author: Greig, Jenny A., Martins, Kelly M., Breton, Camilo, Lamontagne, R. Jason, Zhu, Yanqing, He, Zhenning, White, John, Zhu, Jing-Xu, Chichester, Jessica A., Zheng, Qi, Zhang, Zhe, Bell, Peter, Wang, Lili, Wilson, James M.

Issue&Volume: 2023-11-06

Abstract: The development of liver-based adeno-associated virus (AAV) gene therapies is facing concerns about limited efficiency and durability of transgene expression. We evaluated nonhuman primates following intravenous dosing of AAV8 and AAVrh10 vectors for over 2 years to better define the mechanism(s) of transduction that affect performance. High transduction of non-immunogenic transgenes was achieved, although expression declined over the first 90 days to reach a lower but stable steady state. More than 10% of hepatocytes contained single nuclear domains of vector DNA that persisted despite the loss of transgene expression. Greater reductions in vector DNA and RNA were observed with immunogenic transgenes. Genomic integration of vector sequences, including complex concatemeric structures, were detected in 1 out of 100 cells at broadly distributed loci that were not in proximity to genes associated with hepatocellular carcinoma. Our studies suggest that AAV-mediated transgene expression in primate hepatocytes occurs in two phases: high but short-lived expression from episomal genomes, followed by much lower but stable expression, likely from integrated vectors. Long-term expression of AAV transgenes in macaque liver is traced to integrated vectors.

DOI: 10.1038/s41587-023-01974-7

Source: https://www.nature.com/articles/s41587-023-01974-7

期刊信息

Nature Biotechnology：《自然—生物技术》，创刊于1996年。隶属于施普林格·自然出版集团，最新IF：68.164

官方网址：https://www.nature.com/nbt/

投稿链接：https://mts-nbt.nature.com/cgi-bin/main.plex