title: In Vivo Zinc Finger Nuclease-mediated Targeted Integration of a Glucose-6-phosphatase Transgene Promotes Survival in Mice With Glycogen Storage Disease Type IA.
abstract: Glycogen storage disease type Ia (GSD Ia) is caused by glucose-6-phosphatase (G6Pase) deficiency in association with severe, life-threatening hypoglycemia that necessitates lifelong dietary therapy. Here we show that use of a zinc-finger nuclease (ZFN) targeted to the ROSA26 safe harbor locus and a ROSA26-targeting vector containing a G6PC donor transgene, both delivered with adeno-associated virus (AAV) vectors, markedly improved survival of G6Pase knockout (G6Pase-KO) mice compared with mice receiving the donor vector alone (P < 0.04). Furthermore, transgene integration has been confirmed by sequencing in the majority of the mice treated with both vectors. Targeted alleles were 4.6-fold more common in livers of mice with GSD Ia, as compared with normal littermates, at 8 months following vector administration (P < 0.02). This suggests a selective advantage for vector-transduced hepatocytes following ZFN-mediated integration of the G6Pase vector. A short-term experiment also showed that 3-month-old mice receiving the ZFN had significantly-improved biochemical correction, in comparison with mice that received the donor vector alone. These data suggest that the use of ZFNs to drive integration of G6Pase at a safe harbor locus might improve vector persistence and efficacy, and lower mortality in GSD Ia.
authors: Department of Pediatrics, Division of Medical Genetics, Duke University Medical Center, Durham, North Carolina, USA.|Department of Molecular Genetics and Microbiology, Duke University, Durham, North Carolina, USA.
authors: Dustin J
authors: DJ
authors: Landau
authors: Department of Pediatrics, Division of Medical Genetics, Duke University Medical Center, Durham, North Carolina, USA.|Division of Laboratory Animal Resources, Duke University Medical Center, Durham, North Carolina, USA.
authors: Elizabeth Drake
authors: ED
authors: Brooks
authors: Department of Biomedical Engineering, Duke University, Durham, North Carolina, USA.|Current address: Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
authors: Pablo
authors: P
authors: Perez-Pinera
authors: Department of Pediatrics, Division of Medical Genetics, Duke University Medical Center, Durham, North Carolina, USA.|Current address: Michael E. DeBakey Department of Surgery, Division of Cardiothoracic Surgery, CHI St. Luke's Health-Baylor St. Luke's Medical Center, Houston, Texas, USA.
authors: Hiruni
authors: H
authors: Amarasekara
authors: Department of Molecular Genetics and Microbiology, Duke University, Durham, North Carolina, USA.
authors: Adam
authors: A
authors: Mefferd
authors: Department of Pediatrics, Division of Medical Genetics, Duke University Medical Center, Durham, North Carolina, USA.
authors: Songtao
authors: S
authors: Li
authors: Department of Pediatrics, Division of Medical Genetics, Duke University Medical Center, Durham, North Carolina, USA.
authors: Andrew
authors: A
authors: Bird
authors: Department of Biomedical Engineering, Duke University, Durham, North Carolina, USA.
authors: Charles A
authors: CA
authors: Gersbach
authors: Department of Pediatrics, Division of Medical Genetics, Duke University Medical Center, Durham, North Carolina, USA.|Department of Molecular Genetics and Microbiology, Duke University, Durham, North Carolina, USA.
authors: Dwight D
authors: DD
authors: Koeberl
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D004720:Endonucleases / Q000737:chemistry / Q000378:metabolism*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage; D005952:Glucose-6-Phosphatase / Q000235:genetics*; D005953:Glycogen Storage Disease Type I / Q000235:genetics / Q000628:therapy*; D051379:Mice; D022661:RNA, Untranslated / Q000235:genetics*; D016019:Survival Analysis; D016896:Treatment Outcome; D016335:Zinc Fingers
chemical_list: C110853:Gt(ROSA)26Sor non-coding RNA, mouse; D022661:RNA, Untranslated; D004720:Endonucleases; D005952:Glucose-6-Phosphatase