Department of Medicine, Division of Gastroenterology, Hepatology and Nutrition, University of Minnesota Medical School, Minneapolis, Minnesota 55455, USA.
Department of Medicine, Division of Gastroenterology, Hepatology and Nutrition, University of Minnesota Medical School, Minneapolis, Minnesota 55455, USA; Department of Genetics, Cell Biology and Development, University of Minnesota Medical School, Minneapolis, Minnesota 55455 USA.
Drug Discov Today Technol. 2019 Dec;34:9-19. doi: 10.1016/j.ddtec.2020.11.001. Epub 2020 Nov 22.
The large number of monogenic metabolic disorders originating in the liver poses a unique opportunity for development of gene therapy modalities to pursue curative approaches. Various disorders have been successfully treated via liver-directed gene therapy, though most of the advances have been in animal models, with only limited success in clinical trials. Pre-clinical data in animals using non-viral approaches, including the Sleeping Beauty transposon system, are discussed. The various advances with viral vectors for liver-directed gene therapy are also a focus of this review, including retroviral, adenoviral, recombinant adeno-associated viral, and SV40 vectors. Genome editing techniques, including zinc finger nucleases, transcription activator-like effector nucleases and clustered regularly interspaced short palindromic repeats (CRISPR), are also described. Further, the various controversies in the field with regards to somatic vs. germline editing using CRISPR in humans are explored, while also highlighting the myriad of preclinical advances. Lastly, newer technologies are reviewed, including base editing and prime editing, which use CRISPR with exciting adjunctive properties to avoid double-stranded breaks and thus the recruitment of endogenous repair mechanisms. While encouraging results have been achieved recently, there are still significant challenges to overcome prior to the broad use of vector-based and genome editing techniques in the clinical arena. As these technologies mature, the promise of a cure for many disabling inherited metabolic disorders is within reach, and urgently needed.
大量源于肝脏的单基因代谢紊乱为开发基因治疗方法提供了独特的机会，以寻求根治方法。通过肝靶向基因治疗已经成功治疗了各种疾病，尽管大多数进展都在动物模型中，临床试验中只有有限的成功。本文讨论了使用非病毒方法（包括睡眠美人转座子系统）在动物中的临床前数据。本文还重点介绍了用于肝靶向基因治疗的各种病毒载体的进展，包括逆转录病毒、腺病毒、重组腺相关病毒和 SV40 载体。还描述了基因组编辑技术，包括锌指核酸酶、转录激活因子样效应核酸酶和簇状规则间隔短回文重复序列 (CRISPR)。此外，还探讨了使用 CRISPR 在人类中进行体细胞与生殖细胞编辑的领域中的各种争议，同时强调了众多临床前进展。最后，本文还回顾了新技术，包括碱基编辑和先导编辑，它们使用具有令人兴奋的附加特性的 CRISPR 来避免双链断裂，从而避免招募内源性修复机制。虽然最近取得了令人鼓舞的结果，但在广泛应用基于载体和基因组编辑技术于临床领域之前，仍有许多重大挑战需要克服。随着这些技术的成熟，许多使人丧失能力的遗传性代谢紊乱的治愈希望就在眼前，这是迫切需要的。
