title: Curative ex vivo liver-directed gene therapy in a pig model of hereditary tyrosinemia type 1.
abstract: We tested the hypothesis that ex vivo hepatocyte gene therapy can correct the metabolic disorder in fumarylacetoacetate hydrolase-deficient (Fah(-/-)) pigs, a large animal model of hereditary tyrosinemia type 1 (HT1). Recipient Fah(-/-) pigs underwent partial liver resection and hepatocyte isolation by collagenase digestion. Hepatocytes were transduced with one or both of the lentiviral vectors expressing the therapeutic Fah and the reporter sodium-iodide symporter (Nis) genes under control of the thyroxine-binding globulin promoter. Pigs received autologous transplants of hepatocytes by portal vein infusion. After transplantation, the protective drug 2-(2-nitro-4-trifluoromethylbenzyol)-1,3 cyclohexanedione (NTBC) was withheld from recipient pigs to provide a selective advantage for expansion of corrected FAH(+) cells. Proliferation of transplanted cells, assessed by both immunohistochemistry and noninvasive positron emission tomography imaging of NIS-labeled cells, demonstrated near-complete liver repopulation by gene-corrected cells. Tyrosine and succinylacetone levels improved to within normal range, demonstrating complete correction of tyrosine metabolism. In addition, repopulation of the Fah(-/-) liver with transplanted cells inhibited the onset of severe fibrosis, a characteristic of nontransplanted Fah(-/-) pigs. This study demonstrates correction of disease in a pig model of metabolic liver disease by ex vivo gene therapy. To date, ex vivo gene therapy has only been successful in small animal models. We conclude that further exploration of ex vivo hepatocyte genetic correction is warranted for clinical use.
authors.affiliation: Department of Surgery, Mayo Clinic, Rochester, MN 55905, USA. Department of Molecular Medicine, Mayo Clinic, Rochester, MN 55905, USA. hickey.raymond@mayo.edu.
authors.forename: Raymond D
authors.initials: RD
authors.lastname: Hickey
authors.affiliation: Department of Surgery, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: Shennen A
authors.initials: SA
authors.lastname: Mao
authors.affiliation: Department of Surgery, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: Jaime
authors.initials: J
authors.lastname: Glorioso
authors.affiliation: Department of Surgery, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: Faysal
authors.initials: F
authors.lastname: Elgilani
authors.affiliation: Brami Biomedical Inc., Coon Rapids, MN 55433, USA.
authors.forename: Bruce
authors.initials: B
authors.lastname: Amiot
authors.affiliation: Department of Surgery, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: Harvey
authors.initials: H
authors.lastname: Chen
authors.affiliation: Division of Laboratory Genetics, Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: Piero
authors.initials: P
authors.lastname: Rinaldo
authors.affiliation: Department of Comparative Medicine, Mayo Clinic, Scottsdale, AZ 85259, USA.
authors.forename: Ronald
authors.initials: R
authors.lastname: Marler
authors.affiliation: Department of Radiology, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: Huailei
authors.initials: H
authors.lastname: Jiang
authors.affiliation: Department of Radiology, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: Timothy R
authors.initials: TR
authors.lastname: DeGrado
authors.affiliation: Department of Radiology, Mayo Clinic, Rochester, MN 55905, USA. Imanis Life Sciences, Rochester, MN 55902, USA.
authors.forename: Lukkana
authors.initials: L
authors.lastname: Suksanpaisan
authors.affiliation: Department of Radiology, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: Michael K
authors.initials: MK
authors.lastname: O'Connor
authors.affiliation: Division of Pediatric Gastroenterology, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: Brittany L
authors.initials: BL
authors.lastname: Freeman
authors.affiliation: Division of Pediatric Gastroenterology, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: Samar H
authors.initials: SH
authors.lastname: Ibrahim
authors.affiliation: Department of Molecular Medicine, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: Kah Whye
authors.initials: KW
authors.lastname: Peng
authors.affiliation: Department of Molecular and Medical Genetics and Department of Pediatrics, Oregon Health and Science University, Portland, OR 97239, USA.
authors.forename: Cary O
authors.initials: CO
authors.lastname: Harding
authors.affiliation: Histocompatibility Laboratory, Gift of Life Michigan, Ann Arbor, MI 48108, USA.
authors.forename: Chak-Sum
authors.initials: CS
authors.lastname: Ho
authors.affiliation: Papé Family Pediatric Research Institute, Oregon Health and Science University, Portland, OR 97239, USA.
authors.forename: Markus
authors.initials: M
authors.lastname: Grompe
authors.affiliation: Department of Molecular Medicine, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Ikeda
authors.affiliation: Department of Surgery, Mayo Clinic, Rochester, MN 55905, USA. Midwest Fetal Care Center, Children's Hospitals and Clinics of Minnesota, Minneapolis, MN 55404, USA.
authors.forename: Joseph B
authors.initials: JB
authors.lastname: Lillegard
authors.affiliation: Department of Molecular Medicine, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Russell
authors.affiliation: Department of Surgery, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: Scott L
authors.initials: SL
authors.lastname: Nyberg
mesh_terms: D000818:Animals; D003512:Cyclohexanones / Q000494:pharmacology; D004195:Disease Models, Animal; D015316:Genetic Therapy / Q000379:methods*; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism; D006867:Hydrolases / Q000235:genetics / Q000378:metabolism; D007150:Immunohistochemistry; D008099:Liver / Q000378:metabolism*; D009579:Nitrobenzoates / Q000494:pharmacology; D013552:Swine; D014184:Transplantation, Homologous; D020176:Tyrosinemias / Q000201:enzymology / Q000235:genetics / Q000378:metabolism* / Q000628:therapy*
chemical_list: D003512:Cyclohexanones; D009579:Nitrobenzoates; D006867:Hydrolases; C020156:fumarylacetoacetase; C077073:nitisinone