Virus-Host Interaction Group, Department of Infectious Diseases/Virology, Heidelberg University Hospital, Cluster of Excellence CellNetworks, Heidelberg, 69120, Germany.
BioQuant, University of Heidelberg, Heidelberg, 69120, Germany.
Nat Commun. 2018 Jul 9;9(1):2651. doi: 10.1038/s41467-018-05059-x.
In vivo reprogramming of somatic cells into induced pluripotent stem cells (iPSC) holds vast potential for basic research and regenerative medicine. However, it remains hampered by a need for vectors to express reprogramming factors (Oct-3/4, Klf4, Sox2, c-Myc; OKSM) in selected organs. Here, we report OKSM delivery vectors based on pseudotyped Adeno-associated virus (AAV). Using the AAV-DJ capsid, we could robustly reprogram mouse embryonic fibroblasts with low vector doses. Swapping to AAV8 permitted to efficiently reprogram somatic cells in adult mice by intravenous vector delivery, evidenced by hepatic or extra-hepatic teratomas and iPSC in the blood. Notably, we accomplished full in vivo reprogramming without c-Myc. Most iPSC generated in vitro or in vivo showed transcriptionally silent, intronic or intergenic vector integration, likely reflecting the increased host genome accessibility during reprogramming. Our approach crucially advances in vivo reprogramming technology, and concurrently facilitates investigations into the mechanisms and consequences of AAV persistence.
体细胞重编程为诱导多能干细胞（iPSC）在基础研究和再生医学中有巨大的潜力。然而，它仍然受到需要载体在选定的器官中表达重编程因子（Oct-3/4、Klf4、Sox2、c-Myc；OKSM）的限制。在这里，我们报告了基于假型腺相关病毒（AAV）的 OKSM 传递载体。使用 AAV-DJ 衣壳，我们可以用低剂量的载体有效地重编程小鼠胚胎成纤维细胞。通过静脉内载体递送，将 AAV8 替换为 AAV8 可以有效地在成年小鼠中重编程体细胞，这可以通过肝内或肝外畸胎瘤和血液中的 iPSC 来证明。值得注意的是，我们在没有 c-Myc 的情况下完成了完全的体内重编程。大多数在体外或体内生成的 iPSC 显示转录沉默、内含子或基因间载体整合，这可能反映了在重编程过程中宿主基因组的可及性增加。我们的方法极大地推进了体内重编程技术的发展，同时也促进了对 AAV 持续性的机制和后果的研究。
