Kim Jae-Jun, Kurial Simone N T, Choksi Pervinder K, Nunez Miguel, Lunow-Luke Tyler, Bartel Jan, Driscoll Julia, Her Chris L, Dhillon Simaron, Yue William, Murti Abhishek, Mao Tin, Ramos Julian N, Tiyaboonchai Amita, Grompe Markus, Mattis Aras N, Syed Shareef M, Wang Bruce M, Maher Jacquelyn J, Roll Garrett R, Willenbring Holger
Department of Surgery, University of California, San Francisco, San Francisco, CA, USA.
Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA, USA.
Nat Biotechnol. 2025 Jan 29. doi: 10.1038/s41587-024-02523-6.
Therapeutic efficacy and safety of adeno-associated virus (AAV) liver gene therapy depend on capsid choice. To predict AAV capsid performance under near-clinical conditions, we established side-by-side comparison at single-cell resolution in human livers maintained by normothermic machine perfusion. AAV-LK03 transduced hepatocytes much more efficiently and specifically than AAV5, AAV8 and AAV6, which are most commonly used clinically, and AAV-NP59, which is better at transducing human hepatocytes engrafted in immune-deficient mice. AAV-LK03 preferentially transduced periportal hepatocytes in normal liver, whereas AAV5 targeted pericentral hepatocytes in steatotic liver. AAV5 and AAV8 transduced liver sinusoidal endothelial cells as efficiently as hepatocytes. AAV capsid and steatosis influenced vector episome formation, which determines gene therapy durability, with AAV5 delaying concatemerization. Our findings inform capsid choice in clinical AAV liver gene therapy, including consideration of disease-relevant hepatocyte zonation and effects of steatosis, and facilitate the development of AAV capsids that transduce hepatocytes or other therapeutically relevant cell types in the human liver with maximum efficiency and specificity.
腺相关病毒（AAV）肝脏基因治疗的疗效和安全性取决于衣壳的选择。为了预测AAV衣壳在接近临床条件下的性能，我们在常温机器灌注维持的人肝脏中以单细胞分辨率建立了并排比较。与临床上最常用的AAV5、AAV8和AAV6，以及在免疫缺陷小鼠中植入的人肝细胞转导效果更好的AAV-NP59相比，AAV-LK03转导肝细胞的效率和特异性要高得多。AAV-LK03优先转导正常肝脏中的门静脉周围肝细胞，而AAV5靶向脂肪变性肝脏中的中央周围肝细胞。AAV5和AAV8转导肝窦内皮细胞的效率与肝细胞相同。AAV衣壳和脂肪变性影响载体附加体的形成，而载体附加体的形成决定了基因治疗的持久性，其中AAV5会延迟多联体的形成。我们的研究结果为临床AAV肝脏基因治疗中的衣壳选择提供了参考，包括考虑与疾病相关的肝细胞分区和脂肪变性的影响，并有助于开发能够以最高效率和特异性转导人肝脏中的肝细胞或其他治疗相关细胞类型的AAV衣壳。
