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担忧。我们对静脉注射 AAV8 和 AAVrh10 载体超过 2 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。实现了非免疫原性转基因的高转导，尽管表达在最初的 90 天内下降，达到较低但稳定的稳态。超过 10% 的肝细胞含有载体 DNA 的单核结构域，尽管转基因表达丧失，但该结构域仍然存在。使用免疫原性转基因观察到载体 DNA 和 RNA 的更大减少。在不接近肝细胞癌相关基因的广泛分布的基因座中，在十分之一的细胞中检测到载体序列的基因组整合，包括复杂的多联体结构。我们的研究表明，灵长类动物肝细胞中 AAV 介导的转基因表达分两个阶段：来自游离基因组的高但短暂的表达，随后是低得多但稳定的表达，可能来自整合载体。© 2023。作者。

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.© 2023. The Author(s).