CRISPR/Cas9-mediated gene correction in newborn rabbits with hereditary tyrosinemia type I [DeepSeq2]

GSE142721 CRISPR/Cas9-mediated gene correction in newborn rabbits with hereditary tyrosinemia type I [DeepSeq2]: Contributors : Nan Li ; Shixue Gou ; Jiaowei Wang ; Quanjun Zhang ; Jingke Xie ; Li Li ; Qin Jin ; Ouyang Zhen ; Fangbing Chen ; Weikai Ge ; Hui Shi ; Yanhui Liang ; Huang Xingyun ; Zhenpeng Zhuang ; Xiaozhu Zhao ; Meng Lian ; Yinghua Ye ; Longquan Quan ; Han Wu ; Kepin Wang ; Liangxue Lai

Series Type : Genome variation profiling by high throughput sequencing

Organism : Oryctolagus cuniculus

Mutations in the metabolic enzyme fumarylacetoacetate hydrolase (FAH) cause hereditary tyrosinemia type I (HT1) in human. HT1 patients present acute and irreversible liver and kidney damage during infancy. CRISPR/Cas9-medated precise correction of disease-causing mutations in the liver of infant may provide a promising approach for the treatment of monogenetic liver metabolic disorders. However, to date, all previous precise gene therapy studies were conducted in adult HT1 rodent models (mice and rats), which are not able to recover irreversible pathological changes and result in less treatment effectiveness. In addition, rodent animals have very tiny livers comparing with that of humans, and HT1 rodent models do not authentically recapitulate some symptoms of human patients such as kidney damage. Therefore, to achieve the data which could be more translationable to medical practice of HT1 disease treatment by correcting gene mutation of hepatocytes, here, we chose a HT1 rabbit model, owning relatively large livers, and displaying liver and kidney damage as human patients, as the subject to test the gene therapy effectiveness starting from newborn stage. By delivery CRISPR/Cas9 system and donor templates to the livers by ear vein injection of adeno-associated virus (AAV), HT1 rabbits could be rescued the lethal phenotypes and were able to grow up to adult normally without NTBC treatment and give birth to offspring. In the livers of AAV-treated HT1 rabbits, both HDR-mediated and NHEJ-mediated gene corrections were occurred with the efficiencies ranged from 3.30% to 8.58%. Gene corrected HT1 rabbits showed normal structure and function of both livers and kidneys. This study in rabbits provides useful large-animal preclinical data to treat genetic metabolic disorders affecting the liver with gene therapy.