Reprogramming metabolic pathways in vivo with CRISPR/Cas9 genome editing to treat hereditary tyrosinaemia

Nat Commun. 2016 Aug 30:7:12642. doi: 10.1038/ncomms12642.

Abstract

Many metabolic liver disorders are refractory to drug therapy and require orthotopic liver transplantation. Here we demonstrate a new strategy, which we call metabolic pathway reprogramming, to treat hereditary tyrosinaemia type I in mice; rather than edit the disease-causing gene, we delete a gene in a disease-associated pathway to render the phenotype benign. Using CRISPR/Cas9 in vivo, we convert hepatocytes from tyrosinaemia type I into the benign tyrosinaemia type III by deleting Hpd (hydroxyphenylpyruvate dioxigenase). Edited hepatocytes (Fah(-/-)/Hpd(-/-)) display a growth advantage over non-edited hepatocytes (Fah(-/-)/Hpd(+/+)) and, in some mice, almost completely replace them within 8 weeks. Hpd excision successfully reroutes tyrosine catabolism, leaving treated mice healthy and asymptomatic. Metabolic pathway reprogramming sidesteps potential difficulties associated with editing a critical disease-causing gene and can be explored as an option for treating other diseases.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics*
  • Cell Line
  • Cell Proliferation / genetics
  • Cyclohexanones / therapeutic use
  • Disease Models, Animal
  • Enzyme Inhibitors / therapeutic use
  • Exons / genetics
  • Gene Editing / methods*
  • Genetic Therapy / methods*
  • Hepatocytes / metabolism
  • Humans
  • Hydrolases / genetics
  • Liver / cytology
  • Liver / pathology
  • Metabolic Networks and Pathways / genetics*
  • Mice
  • Mice, Inbred Strains
  • Mice, Knockout
  • Nitrobenzoates / therapeutic use
  • Oxidoreductases / antagonists & inhibitors
  • Oxidoreductases / genetics
  • Oxidoreductases / metabolism
  • Phenotype
  • Tyrosinemias / genetics*
  • Tyrosinemias / metabolism
  • Tyrosinemias / pathology
  • Tyrosinemias / therapy

Substances

  • Cyclohexanones
  • Enzyme Inhibitors
  • Hpd protein, mouse
  • Nitrobenzoates
  • Oxidoreductases
  • Hydrolases
  • fumarylacetoacetase
  • nitisinone