CRISPR/Cas9-Mediated Gene Correction in Newborn Rabbits with Hereditary Tyrosinemia Type I

• Published in: Molecular therapy Vol. 29; no. 3; pp. 1001 - 1015
• Main Authors: Li, Nan, Gou, Shixue, Wang, Jiaowei, Zhang, Quanjun, Huang, Xingyun, Xie, Jingke, Li, Li, Jin, Qin, Ouyang, Zhen, Chen, Fangbing, Ge, Weikai, Shi, Hui, Liang, Yanhui, Zhuang, Zhenpeng, Zhao, Xiaozhu, Lian, Meng, Ye, Yinghua, Quan, Longquan, Wu, Han, Lai, Liangxue, Wang, Kepin
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 03.03.2021; American Society of Gene & Cell Therapy
• Subjects: adeno-associated virus; Animals; Animals, Newborn; CRISPR-Cas Systems; CRISPR-Cas9-mediated precise gene therapy; Dependovirus - genetics; Disease Models, Animal; DNA End-Joining Repair; Female; Gene Editing; Gene Expression Regulation; Genetic Therapy; Genetic Vectors - administration & dosage; hereditary tyrosinemia type I; Hydrolases - genetics; Kidney - metabolism; Liver - metabolism; Male; Original; rabbit models; Rabbits; RNA-Seq; Tyrosinemias - genetics; Tyrosinemias - pathology; Tyrosinemias - therapy; hereditary tyrosinemia type I; adeno-associated virus; rabbit models; CRISPR-Cas9-mediated precise gene therapy
• ISSN: 1525-0016; 1525-0024
• DOI: 10.1016/j.ymthe.2020.11.023

Patients with hereditary tyrosinemia type I (HT1) present acute and irreversible liver and kidney damage during infancy. CRISPR-Cas9-mediated gene correction during infancy may provide a promising approach to treat patients with HT1. However, all previous studies were performed on adult HT1 rodent models, which cannot authentically recapitulate some symptoms of human patients. The efficacy and safety should be verified in large animals to translate precise gene therapy to clinical practice. Here, we delivered CRISPR-Cas9 and donor templates via adeno-associated virus to newborn HT1 rabbits. The lethal phenotypes could be rescued, and notably, these HT1 rabbits reached adulthood normally without 2-(2-nitro-4-trifluoromethylbenzyol)-1,3 cyclohexanedione administration and even gave birth to offspring. Adeno-associated virus (AAV)-treated HT1 rabbits displayed normal liver and kidney structures and functions. Homology-directed repair-mediated precise gene corrections and non-homologous end joining-mediated out-of-frame to in-frame corrections in the livers were observed with efficiencies of 0.90%–3.71% and 2.39%–6.35%, respectively, which appeared to be sufficient to recover liver function and decrease liver and kidney damage. This study provides useful large-animal preclinical data for rescuing hepatocyte-related monogenetic metabolic disorders with precise gene therapy. [Display omitted] CRISPR-Cas9-mediated precise FAH gene correction is a promising strategy to treat hereditary tyrosinemia type I (HT1). In this study, Wang and colleagues rescued the lethal phenotypes and recovered the liver and kidney function of HT1 rabbits by CRISPR-Cas9-mediated precise FAH gene correction. This study provides useful large-animal preclinical data for rescuing hepatocyte-related monogenetic metabolic disorders with precise gene therapy.