title: Reprogramming metabolic pathways in vivo with CRISPR/Cas9 genome editing to treat hereditary tyrosinaemia.
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.
authors: Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas 77030, USA.|Center for Stem Cells and Regenerative Medicine, Baylor College of Medicine, Houston, Texas 77030, USA.|Graduate Program, Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas 77030, USA.
authors: Francis P
authors: FP
authors: Pankowicz
authors: Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas 77030, USA.|Center for Stem Cells and Regenerative Medicine, Baylor College of Medicine, Houston, Texas 77030, USA.
authors: Mercedes
authors: M
authors: Barzi
authors: Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas 77030, USA.|Center for Stem Cells and Regenerative Medicine, Baylor College of Medicine, Houston, Texas 77030, USA.
authors: Xavier
authors: X
authors: Legras
authors: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA.
authors: Leroy
authors: L
authors: Hubert
authors: Department of Pediatrics, Texas Children's Hospital, Houston, Texas, USA.
authors: Tian
authors: T
authors: Mi
authors: Graduate Program in Translational Biology and Molecular Medicine, Baylor College of Medicine, Houston, Texas 77030, USA.
authors: Julie A
authors: 0000-0002-6012-3948
authors: JA
authors: Tomolonis
authors: Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas 77030, USA.|Center for Stem Cells and Regenerative Medicine, Baylor College of Medicine, Houston, Texas 77030, USA.
authors: Milan
authors: M
authors: Ravishankar
authors: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA.
authors: Qin
authors: Q
authors: Sun
authors: Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas 77030, USA.|Center for Stem Cells and Regenerative Medicine, Baylor College of Medicine, Houston, Texas 77030, USA.|Graduate Program, Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas 77030, USA.|McNair Medical Institute, Houston, Texas, USA.
authors: Diane
authors: D
authors: Yang
authors: Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas 77030, USA.|Center for Stem Cells and Regenerative Medicine, Baylor College of Medicine, Houston, Texas 77030, USA.|Graduate Program, Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas 77030, USA.|Graduate Program in Translational Biology and Molecular Medicine, Baylor College of Medicine, Houston, Texas 77030, USA.|McNair Medical Institute, Houston, Texas, USA.|Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas, USA.|Program in Developmental Biology, Baylor College of Medicine, Houston, Texas 77030, USA.|Dan L. Duncan Cancer Center, Baylor College of Medicine, Houston, Texas 77030, USA.
authors: Malgorzata
authors: M
authors: Borowiak
authors: Department of Pediatrics, Texas Children's Hospital, Houston, Texas, USA.|Dan L. Duncan Cancer Center, Baylor College of Medicine, Houston, Texas 77030, USA.
authors: Pavel
authors: P
authors: Sumazin
authors: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA.
authors: Sarah H
authors: SH
authors: Elsea
authors: Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas 77030, USA.|Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas, USA.
authors: Beatrice
authors: B
authors: Bissig-Choisat
authors: Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas 77030, USA.|Center for Stem Cells and Regenerative Medicine, Baylor College of Medicine, Houston, Texas 77030, USA.|Graduate Program, Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas 77030, USA.|Graduate Program in Translational Biology and Molecular Medicine, Baylor College of Medicine, Houston, Texas 77030, USA.|Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas, USA.|Program in Developmental Biology, Baylor College of Medicine, Houston, Texas 77030, USA.|Dan L. Duncan Cancer Center, Baylor College of Medicine, Houston, Texas 77030, USA.
authors: Karl-Dimiter
authors: KD
authors: Bissig
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems / Q000235:genetics*; D002460:Cell Line; D049109:Cell Proliferation / Q000235:genetics; D003512:Cyclohexanones / Q000627:therapeutic use; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000627:therapeutic use; D005091:Exons / Q000235:genetics; D000072669:Gene Editing / Q000379:methods*; D015316:Genetic Therapy / Q000379:methods*; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D006867:Hydrolases / Q000235:genetics; D008099:Liver / Q000166:cytology / Q000473:pathology; D053858:Metabolic Networks and Pathways / Q000235:genetics*; D051379:Mice; D008815:Mice, Inbred Strains; D018345:Mice, Knockout; D009579:Nitrobenzoates / Q000627:therapeutic use; D010088:Oxidoreductases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D020176:Tyrosinemias / Q000235:genetics* / Q000378:metabolism / Q000473:pathology / Q000628:therapy
chemical_list: D003512:Cyclohexanones; D004791:Enzyme Inhibitors; C488911:Hpd protein, mouse; D009579:Nitrobenzoates; D010088:Oxidoreductases; D006867:Hydrolases; C020156:fumarylacetoacetase; C077073:nitisinone