title: Rescue of bilirubin-induced neonatal lethality in a mouse model of Crigler-Najjar syndrome type I by AAV9-mediated gene transfer.
abstract: Crigler-Najjar type I (CNI) syndrome is a recessively inherited disorder characterized by severe unconjugated hyperbilirubinemia caused by uridine diphosphoglucuronosyltransferase 1A1 (UGT1A1) deficiency. The disease is lethal due to bilirubin-induced neurological damage unless phototherapy is applied from birth. However, treatment becomes less effective during growth, and liver transplantation is required. To investigate the pathophysiology of the disease and therapeutic approaches in mice, we generated a mouse model by introducing a premature stop codon in the UGT1a1 gene, which results in an inactive enzyme. Homozygous mutant mice developed severe jaundice soon after birth and died within 11 d, showing significant cerebellar alterations. To rescue neonatal lethality, newborns were injected with a single dose of adeno-associated viral vector 9 (AAV9) expressing the human UGT1A1. Gene therapy treatment completely rescued all AAV-treated mutant mice, accompanied by lower plasma bilirubin levels and normal brain histology and motor coordination. Our mouse model of CNI reproduces genetic and phenotypic features of the human disease. We have shown, for the first time, the full recovery of the lethal effects of neonatal hyperbilirubinemia. We believe that, besides gene-addition-based therapies, our mice could represent a very useful model to develop and test novel technologies based on gene correction by homologous recombination.
authors.affiliation: International Centre for Genetic Engineering and Biotechnology, Padriciano 99, I-34149 Trieste, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Bortolussi
authors.forename: Lorena
authors.initials: L
authors.lastname: Zentilin
authors.forename: Gabriele
authors.initials: G
authors.lastname: Baj
authors.forename: Pablo
authors.initials: P
authors.lastname: Giraudi
authors.forename: Cristina
authors.initials: C
authors.lastname: Bellarosa
authors.forename: Mauro
authors.initials: M
authors.lastname: Giacca
authors.forename: Claudio
authors.initials: C
authors.lastname: Tiribelli
authors.forename: Andrés F
authors.initials: AF
authors.lastname: Muro
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001663:Bilirubin / Q000097:blood; D015152:Blotting, Northern; D015153:Blotting, Western; D002531:Cerebellum / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D003414:Crigler-Najjar Syndrome / Q000201:enzymology / Q000235:genetics* / Q000401:mortality; D000229:Dependovirus / Q000145:classification / Q000235:genetics; D004195:Disease Models, Animal; D015971:Gene Expression Regulation, Enzymologic; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D014453:Glucuronosyltransferase / Q000172:deficiency / Q000235:genetics*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D018482:Muscle, Skeletal / Q000201:enzymology / Q000378:metabolism; D009154:Mutation; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015996:Survival Rate; D000099148:UGT1A1 Enzyme
chemical_list: D001663:Bilirubin; D014453:Glucuronosyltransferase; D000099148:UGT1A1 Enzyme
references.citation: J Biol Chem. 2008 Mar 21;283(12):7901-11
references.pmid: 18180294
references.citation: J Biol Chem. 1984 May 10;259(9):5500-6
references.pmid: 6715357
references.citation: Behav Brain Res. 2005 Jun 3;161(1):31-8
references.pmid: 15904707
references.citation: Clin Chem. 1985 Nov;31(11):1779-89
references.pmid: 4053346
references.citation: J Gene Med. 2006 Aug;8(8):990-7
references.pmid: 16685745
references.citation: Nature. 2011 Jun 26;475(7355):217-21
references.pmid: 21706032
references.citation: Clin Perinatol. 2006 Jun;33(2):387-410
references.pmid: 16765731
references.citation: Mol Ther. 2006 Jun;13(6):1085-92
references.pmid: 16581301
references.citation: J Cell Biol. 2003 Jul 7;162(1):149-60
references.pmid: 12847088
references.citation: J Gene Med. 2009 Aug;11(8):689-96
references.pmid: 19455564
references.citation: Methods Mol Biol. 2009;530:423-33
references.pmid: 19266333
references.citation: Hum Mutat. 2000 Oct;16(4):297-306
references.pmid: 11013440
references.citation: Mol Ther. 2005 Nov;12(5):860-6
references.pmid: 16019265
references.citation: Mol Ther. 2004 Nov;10(5):844-54
references.pmid: 15509502
references.citation: Pediatrics. 1952 Aug;10(2):169-80
references.pmid: 12983120
references.citation: Nagoya J Med Sci. 1993 Mar;55(1-4):11-21
references.pmid: 8247099
references.citation: Anal Biochem. 1985 Oct;150(1):76-85
references.pmid: 3843705
references.citation: J Virol. 2001 Dec;75(24):12279-87
references.pmid: 11711618
references.citation: Eur J Neurosci. 2005 Jul;22(1):21-7
references.pmid: 16029192
references.citation: Arch Dis Child. 1975 Nov;50(11):833-6
references.pmid: 1108807
references.citation: Biochim Biophys Acta. 1998 Sep 30;1407(3):173-84
references.pmid: 9748558
references.citation: Neurosci Res. 1989 Feb;6(3):209-15
references.pmid: 2710423
references.citation: Arch Biochem Biophys. 2002 Sep 15;405(2):163-9
references.pmid: 12220528
references.citation: Genet Vaccines Ther. 2008 Sep 23;6:13
references.pmid: 18811960
references.citation: Proc Natl Acad Sci U S A. 1996 Nov 12;93(23):13090-5
references.pmid: 8917549
references.citation: Curr Gene Ther. 2009 Apr;9(2):72-82
references.pmid: 19355865
references.citation: Adv Vet Sci Comp Med. 1993;37:149-73
references.pmid: 8273513
references.citation: Eur J Pediatr. 2006 May;165(5):306-19
references.pmid: 16435131
references.citation: Proc Natl Acad Sci U S A. 2010 Mar 16;107(11):5024-9
references.pmid: 20194756
references.citation: Dev Dyn. 1996 Oct;207(2):145-56
references.pmid: 8906418
references.citation: Mol Ther. 2006 Jul;14(1):45-53
references.pmid: 16713360
references.citation: Toxicol Lett. 2000 Mar 15;112-113:9-16
references.pmid: 10720707
references.citation: J Neuropathol Exp Neurol. 1967 Jul;26(3):377-96
references.pmid: 6027437
references.citation: J Virol. 2002 Jan;76(2):791-801
references.pmid: 11752169
references.citation: J Biol Chem. 1989 Dec 15;264(35):21302-7
references.pmid: 2512292
references.citation: Neuromolecular Med. 2006;8(4):513-29
references.pmid: 17028373