Abstract
Among inherited diseases of the liver, Crigler–Najjar type 1 disease (CN-1), which results from complete deficiency in bilirubin UDP-glucuronosyltransferase activity (B-UGT1), is an attractive target for gene therapy studies. Hyperbilirubinemic Gunn rats, a model of CN-1, were injected at 2 days of age with lentiviral or oncoretroviral vectors encoding the human B-UGT1. After injection, bilirubinemia was normalized for up to 95 weeks. Bilirubin conjugates were present in the bile, demonstrating liver transduction. PCR and enzyme activity analysis confirmed gene and phenotype correction in liver. We observed that when using a strong viral promoter, a complete correction was achieved with less than 5% of B-UGT1 copy per haploid genome and after a reconstitution of 12% B-UGT1 normal activity. Liver histology remained normal throughout the experiment and tissue distribution analysis revealed preferential hepatocyte transduction after systemic delivery. Finally, no adverse immune response occurred even after induction of nonspecific liver inflammation, suggesting immune ignorance to the therapeutic protein. Our present results document the lifelong safety of gene therapy for CN-1 with retroviral vectors. They offer a better delineation of liver gene correction level required to achieve complete correction of bilirubinemia and pave the way for future clinical application of gene therapy for inherited liver disorders.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Aubert D, Menoret S, Chiari E, Pichard V, Durand S, Tesson L et al. Cytotoxic immune response blunts long-term transgene expression after efficient retroviral-mediated hepatic gene transfer in rat. Mol Ther 2002; 5: 388–396.
Askari FK, Hitomi Y, Mao M, Wilson JM . Complete correction of hyperbilirubinemia in the Gunn rat model of Crigler-Najjar syndrome type I following transient in vivo adenovirus- mediated expression of human bilirubin UDP-glucuronosyltransferase. Gene Therapy 1996; 3: 381–388.
Sauter BV, Parashar B, Chowdhury NR, Kadakol A, Ilan Y, Singh H et al. A replication-deficient rSV40 mediates liver-directed gene transfer and a long-term amelioration of jaundice in gunn rats. Gastroenterology 2000; 119: 1348–1357.
Seppen J, Bakker C, de Jong B, Kunne C, van den Oever K, Vandenberghe K et al. Adeno-associated virus vector serotypes mediate sustained correction of bilirubin UDP glucuronosyltransferase deficiency in rats. Mol Ther 2006; 13: 1085–1092.
Tada K, Chowdhury NR, Neufeld D, Bosma PJ, Heard M, Prasad VR et al. Long-term reduction of serum bilirubin levels in Gunn rats by retroviral gene transfer in vivo. Liver Transplant Surg 1998; 4: 78–88.
Toietta G, Mane VP, Norona WS, Finegold MJ, Ng P, McDonagh AF et al. Lifelong elimination of hyperbilirubinemia in the Gunn rat with a single injection of helper-dependent adenoviral vector. Proc Natl Acad Sci USA 2005; 102: 3930–3935.
Bellodi-Privato M, Aubert D, Pichard V, Myara A, Trivin F, Ferry N . Successful gene therapy of the Gunn rat by in vivo neonatal hepatic gene transfer using murine oncoretroviral vectors. Hepatology 2005; 42: 431–438.
Nguyen TH, Bellodi-Privato M, Aubert D, Pichard V, Myara A, Trono D et al. Therapeutic lentivirus-mediated neonatal in vivo gene therapy in hyperbilirubinemic Gunn rats. Mol Ther 2005; 12: 852–859.
Vigna E, Amendola M, Benedicenti F, Simmons AD, Follenzi A, Naldini L . Efficient Tet-dependent expression of human factor IX in vivo by a new self-regulating lentiviral vector. Mol Ther 2005; 11: 763–775.
Seppen J, van der Rijt R, Looije N, van Til NP, Lamers WH, Oude Elferink RP . Long-term correction of bilirubin UDPglucuronyltransferase deficiency in rats by in utero lentiviral gene transfer. Mol Ther 2003; 8: 593–599.
Tiegs G, Hentschel J, Wendel A . A T cell-dependent experimental liver injury in mice inducible by concanavalin A. J Clin Invest 1992; 90: 196–203.
Ghosh A, Allamarvdasht M, Pan CJ, Sun MS, Mansfield BC, Byrne BJ et al. Long-term correction of murine glycogen storage disease type Ia by recombinant adeno-associated virus-1-mediated gene transfer. Gene Therapy 2006; 13: 321–329.
Ding Z, Georgiev P, Thony B . Administration-route and gender-independent long-term therapeutic correction of phenylketonuria (PKU) in a mouse model by recombinant adeno-associated virus 8 pseudotyped vector-mediated gene transfer. Gene Therapy 2006; 13: 587–593.
Ponder KP, Melniczek JR, Xu L, Weil MA, O'Malley TM, O'Donnell PA et al. Therapeutic neonatal hepatic gene therapy in mucopolysaccharidosis VII dogs. Proc Natl Acad Sci USA 2002; 99: 13102–13107.
Davidoff AM, Gray JT, Ng CY, Zhang Y, Zhou J, Spence Y et al. Comparison of the ability of adeno-associated viral vectors pseudotyped with serotype 2, 5, and 8 capsid proteins to mediate efficient transduction of the liver in murine and nonhuman primate models. Mol Ther 2005; 11: 875–888.
van der Wegen P, Louwen R, Imam AM, Buijs-Offerman RM, Sinaasappel M, Grosveld F et al. Successful treatment of UGT1A1 deficiency in a rat model of Crigler-Najjar disease by intravenous administration of a liver-specific lentiviral vector. Mol Ther 2006; 13: 374–381.
VandenDriessche T, Thorrez L, Naldini L, Follenzi A, Moons L, Berneman Z et al. Lentiviral vectors containing the human immunodeficiency virus type-1 central polypurine tract can efficiently transduce nondividing hepatocytes and antigen-presenting cells in vivo. Blood 2002; 100: 813–822.
Carbonaro DA, Jin X, Petersen D, Wang X, Dorey F, Kil KS et al. In vivo transduction by intravenous injection of a lentiviral vector expressing human ADA into neonatal ADA gene knockout mice: a novel form of enzyme replacement therapy for ADA deficiency. Mol Ther 2006; 13: 1110–1120.
Pfeifer A, Kessler T, Yang M, Baranov E, Kootstra N, Cheresh D et al. Transduction of liver cells by lentiviral vectors: analysis in living animals by fluorescence imaging. Mol Ther 2001; 3: 319–322.
Follenzi A, Sabatino G, Lombardo A, Boccaccio C, Naldini L . Efficient gene delivery and targeted expression to hepatocytes in vivo by improved lentiviral vectors. Hum Gene Therapy 2002; 13: 243–260.
Ohashi K, Park F, Kay MA . Role of hepatocyte direct hyperplasia in lentivirus-mediated liver transduction in vivo. Hum Gene Therapy 2002; 13: 653–663.
Park F, Ohashi K, Chiu W, Naldini L, Kay M . Efficient lentiviral transduction of liver requires cell cycling in vivo. Nat Genet 2000; 24: 49–52.
Fox IJ, Chowdhury JR, Kaufman SS, Goertzen TC, Chowdhury NR, Warkentin PI et al. Treatment of the Crigler-Najjar syndrome type I with hepatocyte transplantation. N Engl J Med 1998; 338: 1422–1426.
Xu L, Gao C, Sands MS, Cai SR, Nichols TC, Bellinger DA et al. Neonatal or hepatocyte growth factor-potentiated adult gene therapy with a retroviral vector results in therapeutic levels of canine factor IX for hemophilia B. Blood 2003; 101: 3924–3932.
Brown BD, Lillicrap D . Dangerous liaisons: the role of 'danger' signals in the immune response to gene therapy. Blood 2002; 100: 1133–1140.
Wiegard C, Frenzel C, Herkel J, Kallen KJ, Schmitt E, Lohse AW . Murine liver antigen presenting cells control suppressor activity of CD4+CD25+ regulatory T cells. Hepatology 2005; 42: 193–199.
Ando K, Guidotti LG, Cerny A, Ishikawa T, Chisari FV . CTL access to tissue antigen is restricted in vivo. J Immunol 1994; 153: 482–488.
Seppen J, van Til NP, van der Rijt R, Hiralall JK, Kunne C, Elferink RP . Immune response to lentiviral bilirubin UDP-glucuronosyltransferase gene transfer in fetal and neonatal rats. Gene Therapy 2006; 13: 672–677.
Bachrich T, Thalhammer T, Jager W, Haslmayer P, Alihodzic B, Bakos S et al. Characterization of autoantibodies against uridine-diphosphate glucuronosyltransferase in patients with inflammatory liver diseases. Hepatology 2001; 33: 1053–1059.
Hacein-Bey-Abina S, Von Kalle C, Schmidt M, McCormack MP, Wulffraat N, Leboulch P et al. LMO2-associated clonal T cell proliferation in two patients after gene therapy for SCID-X1. Science 2003; 302: 415–419.
Oertel M, Rosencrantz R, Chen YQ, Thota PN, Sandhu JS, Dabeva MD et al. Repopulation of rat liver by fetal hepatoblasts and adult hepatocytes transduced ex vivo with lentiviral vectors. Hepatology 2003; 37: 994–1005.
Gunn CH . Hereditary acholuric jaundice in a new mutant strain of rats. J Hered 1938; 29: 137–139.
Zennou V, Petit C, Guetard D, Nerhbass U, Montagnier L, Charneau P . HIV-1 genome nuclear import is mediated by a central DNA flap. Cell 2000; 10: 173–185.
Follenzi A, Ailles L, Bakovic S, Geuna M, Naldini L . Gene transfer by lentiviral vectors is limited by nuclear translocation and rescued by HIV-1 pol sequences. Nat Genet 2000; 25: 217–222.
Zufferey R, Donello JE, Trono D, Hope TJ . Woodchuck hepatitis virus posttranscriptional regulatory element enhances expression of transgenes delivered by retroviral vectors. J Virol 1999; 73: 2886–2892.
Kostic C, Chiodini F, Salmon P, Wiznerowicz M, Deglon N, Hornfeld D et al. Activity analysis of housekeeping promoters using self-inactivating lentiviral vector delivery into the mouse retina. Gene Therapy 2003; 10: 818–821.
Nguyen TH, Oberholzer J, Birraux J, Majno P, Morel P, Trono D . Highly efficient lentiviral vector-mediated transduction of nondividing, fully reimplantable primary hepatocytes. Mol Ther 2002; 6: 199–209.
Ory DS, Neugeboren BA, Mulligan RC . A stable human-derived packaging cell line for production of high titer retrovirus/vesicular stomatitis virus G pseudotypes. Proc Natl Acad Sci USA 1996; 93: 11400–11406.
Muraca M, Blanckaert N . Liquid-chromatographic assay and identification of mono- and diester conjugates of bilirubin in normal serum. Clin Chem 1983; 29: 1767–1771.
Black M, Billing BH, Heirwegh KP . Determination of bilirubin UDP-glucuronyl transferase activity in needle-biopsy specimens of human liver. Clin Chim Acta 1970; 29: 27–35.
Acknowledgements
This work was supported by grants from Association Française contre les Myopathies (AFM) and from the GIS-Maladies Rares. We thank the Lentiviral Vector Production Unit of the Swiss Institute of Technology Lausanne (http://tronolab.epfl.ch) supported by the AFM for providing stocks of lentiviral vectors.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Nguyen, T., Aubert, D., Bellodi-Privato, M. et al. Critical assessment of lifelong phenotype correction in hyperbilirubinemic Gunn rats after retroviral mediated gene transfer. Gene Ther 14, 1270–1277 (2007). https://doi.org/10.1038/sj.gt.3302993
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.gt.3302993