Berreur Estelle, Lazzaroni Giacomo, Roth Cyrill, Zihlmann Marco, Stirn Martina, Matheis Ramona, Xicluna Rebecca, Breous-Nystrom Ekaterina, Roth Adrian B
Roche Pharma Research and Early Development, Roche Innovation Center Basel, CH-4070 Basel, Switzerland.
Institute of Human Biology, Roche Innovation Center Basel, CH-4070 Basel, Switzerland.
Mol Ther Methods Clin Dev. 2025 Apr 14;33(2):101467. doi: 10.1016/j.omtm.2025.101467. eCollection 2025 Jun 12.
Adeno-associated virus (AAV) vectors are widely used in gene therapy, particularly for liver-targeted treatments. However, predicting human-specific outcomes, such as transduction efficiency and hepatotoxicity, remains challenging. Reliable models are urgently needed to bridge the gap between preclinical studies and clinical applications. This study presents the first comparative evaluation of AAV transduction across multiple induced pluripotent stem cell (iPSC)-derived hepatocyte organoid donors, offering a novel platform for assessing vector performance in human liver models. The transduction efficiency and hepatotoxicity of eight AAV serotypes (AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV8, and AAV9) were tested in iPSC-derived liver organoids and hepatic cell lines (HepG2 and HepaRG). AAV6 and AAV8 exhibited the highest transduction efficiency in organoids, while AAV4 and AAV5 were the least effective. Transduction variability was observed across different donors and cell lines. Notably, no significant hepatotoxicity, measured by AST (aspartate aminotransferase) release and viability measurements, was observed, indicating that AAVs do not induce immediate liver damage . This study introduces iPSC-derived hepatocyte organoids as a novel and effective tool for predicting AAV transduction efficiency and safety, with potential to enhance the translation of gene therapies to clinical applications.
腺相关病毒（AAV）载体广泛应用于基因治疗，尤其是肝脏靶向治疗。然而，预测人类特异性结果，如转导效率和肝毒性，仍然具有挑战性。迫切需要可靠的模型来弥合临床前研究与临床应用之间的差距。本研究首次对多种诱导多能干细胞（iPSC）来源的肝细胞类器官供体的AAV转导进行了比较评估，为评估人类肝脏模型中的载体性能提供了一个新平台。在iPSC来源的肝脏类器官和肝细胞系（HepG2和HepaRG）中测试了八种AAV血清型（AAV1、AAV2、AAV3、AAV4、AAV5、AAV6、AAV8和AAV9）的转导效率和肝毒性。AAV6和AAV8在类器官中表现出最高的转导效率，而AAV4和AAV5效果最差。在不同供体和细胞系中观察到转导变异性。值得注意的是，通过AST（天冬氨酸转氨酶）释放和活力测量未观察到明显的肝毒性，表明AAV不会立即引起肝损伤。本研究引入了iPSC来源的肝细胞类器官作为预测AAV转导效率和安全性的一种新型有效工具，具有增强基因治疗向临床应用转化的潜力。
