Abstract
Transgene expression from helper-dependent adenoviral (HD-Ad) vectors is effective and long lasting, but not permanent. Their use is also limited by the host response against capsid proteins that precludes successful gene expression upon readministration. In this report, we test the hypothesis that PEGylation of HD-Ad reduces its toxicity and promotes transgene expression upon readministration. PEGylation did not compromise transduction efficiency in vitro and in vivo and reduced peak serum IL-6 levels two-fold. IL-12 and TNF-α levels were reduced three- and seven-fold, respectively. Thrombocytopenia was not detected in mice treated with the PEGylated vector. Serum transaminases were not significantly elevated in mice treated with either vector. Mice immunized with 1 × 1011 particles of unmodified HD-Ad expressing human alpha-1 antitrypsin (hA1AT) were rechallenged 28 days later with 8 × 1010 particles of unmodified or PEG-conjugated vector expressing beta-galactosidase. Trace levels of beta-galactosidase (52.23±19.2 pg/mg protein) were detected in liver homogenates of mice that received two doses of unmodified HD-Ad. Mice rechallenged with PEGylated HD-Ad produced significant levels of beta-galactosidase (5.1±0.4 × 105 pg/mg protein, P=0.0001). This suggests that PEGylation of HD-Ad vectors may be appropriate for their safe and efficient use in the clinic.
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References
Ginsberg HS . The life and times of adenoviruses. Adv Virus Res 1999; 54: 1–13.
Jaffe HA et al. Adenovirus-mediated in vivo gene transfer and expression in normal rat liver. Nat Genet 1992; 1: 372–378.
Gomez-Foix AM et al. Adenovirus-mediated transfer of the muscle glycogen phosphorylase gene into hepatocytes confers altered regulation of glycogen metabolism. J Biol Chem 1992; 267: 25129–25134.
Nunes FA, Furth EE, Wilson JM, Raper SE . Gene transfer into the liver of nonhuman primates with E1-deleted recombinant adenoviral vectors: safety of readministration. Hum Gene Ther 1999; 10: 2515–2526.
Christ M et al. Modulation of the inflammatory properties and hepatotoxicity of recombinant adenovirus vectors by the viral E4 gene products. Hum Gene Ther 2000; 11: 415–427.
Pastore L et al. Use of a liver-specific promoter reduces immune response to the transgene in adenoviral vectors. Hum Gene Ther 1999; 10: 1773–1781.
Morral N et al. Lethal toxicity, severe endothelial injury, and a threshold effect with high doses of an adenoviral vector in baboons. Hum Gene Ther 2002; 13: 143–154.
Muruve DA, Barnes MJ, Stillman IE, Libermann TA . Adenoviral gene therapy leads to rapid induction of multiple chemokines and acute neutrophil-dependent hepatic injury in vivo. Hum Gene Ther 1999; 10: 965–976.
Lieber A et al. The role of Kupffer cell activation and viral gene expression in early liver toxicity after infusion of recombinant adenovirus vectors. J Virol 1997; 71: 8798–8807.
Zhang Y et al. Acute cytokine response to systemic adenoviral vectors in mice is mediated by dendritic cells and macrophages. Mol Ther 2001; 3: 697–707.
Liu Q, Muruve DA . Molecular basis of the inflammatory response to adenovirus vectors. Gene Therapy 2003; 10: 935–940.
Jooss K, Ertl HC, Wilson JM . Cytotoxic T-lymphocyte target proteins and their major histocompatibility complex class I restriction in response to adenovirus vectors delivered to mouse liver. J Virol 1998; 72: 2945–2954.
Yang Y et al. Immune responses to viral antigens versus transgene product in the elimination of recombinant adenovirus-infected adenovirus hepatocytes in vivo. Gene Therapy 1996; 3: 137–144.
Yang Y, Wilson JM . Clearance of adenovirus-infected hepatocytes by MHC class I-restricted CD4+ CTLAs in vivo. J Immunol 1995; 155: 2564–2570.
Raper SE et al. Fatal systemic inflammatory response syndrome in a ornithine transcarbamylase deficient patient following adenoviral gene transfer. Mol Genet Metab 2003; 80: 148–158.
Schnell MA et al. Activation of innate immunity in nonhuman primates following intraportal administration of adenoviral vectors. Mol Ther 2001; 3: 708–722.
Yang Y, Greenough K, Wilson JM . Transient immune blockade prevents formation of neutralizing antibody to recombinant adenovirus and allows repeated gene transfer to mouse liver. Gene Therapy 1996; 3: 412–420.
Kochanek S, Schiedner G, Volpers C . High-capacity ‘gutless’ adenoviral vectors. Curr Opin Mol Ther 2001; 3: 454–463.
O'Neal WK et al. Toxicity associated with repeated administration of first-generation adenovirus vectors does not occur with a helper-dependent vector. Mol Med 2000; 6: 179–195.
Morral N et al. High doses of a helper-dependent adenoviral vector yield supraphysiological levels of alpha1-antitrypsin with negligible toxicity. Hum Gene Ther 1998; 9: 2709–2716.
Kim IH et al. Lifetime correction of genetic deficiency in mice with a single injection of helper-dependent adenoviral vector. Proc Natl Acad Sci USA 2001; 98: 13282–13287.
Oka K et al. Long-term stable correction of low-density lipoprotein receptor-deficient mice with a helper-dependent adenoviral vector expressing the very low-density lipoprotein receptor. Circulation 2001; 103: 1274–1281.
Schiedner G et al. Selective depletion or blockade of Kupffer cells leads to enhanced and prolonged hepatic transgene expression using high-capacity adenoviral vectors. Mol Ther 2003; 7: 35–43.
Brunetti-Pierri N et al. Acute toxicity after high-dose systemic injection of helper-dependent adenoviral vectors into nonhuman primates. Hum Gene Ther 2004; 15: 35–46.
Muruve DA et al. Helper-dependent adenovirus vectors elicit intact innate but attenuated adaptive host immune responses in vivo. J Virol 2004; 78: 5966–5972.
Parks RJ, Evelegh CM, Graham FL . Use of helper-dependent adenoviral vectors of alternative serotypes permits repeat vector administration. Gene Therapy 1999; 6: 1565–1573.
Croyle MA, Chirmule N, Zhang Y, Wilson JM . PEGylation of E1-deleted adenovirus vectors allows significant gene expression on readministration to liver. Hum Gene Ther 2002; 13: 1887–1900.
Croyle MA, Yu QC, Wilson JM . Development of a rapid method for the PEGylation of adenoviruses with enhanced transduction and improved stability under harsh storage conditions. Hum Gene Ther 2000; 11: 1721–1730.
Sims GEC, Snape TJ . A method for the estimation of polyethylene glycol in plasma protein fractions. Anal Biochem 1980; 107: 60–63.
Liu Q et al. The role of capsid-endothelial interactions in the innate immune response to adenovirus vectors. Hum Gene Ther 2003; 14: 627–643.
Trapnell BC, Shanley TP . Innate immune response to in vivo adenovirus infection. In: Curiel D, Douglas JT (eds). Adenoviral Vectors for Gene Therapy. Academic Press: San Diego, 2002, pp 349–373.
Quimby FW . The Mouse. In: Loeb WF, Quimby FW (eds). The Clinical Chemistry of Laboratory Animals. Taylor and Francis: Philadelphia, 1999, pp 3–33.
Saito T et al. Immune tolerance induced by polyethylene glycol-conjugate of protein antigen: clonal deletion of antigen-specific Th-cells in the thymus. J Biomater Sci Polym Ed 2000; 11: 647–656.
So T et al. Extended blood half-life of monomethoxypolyethylene glycol-conjugated hen lysozyme is a key parameter controlling immunological tolerogenicity. Cell Mol Life Sci 1999; 55: 1187–1194.
Harris JM, Chess RB . Effect of PEGylation on pharmaceuticals. Nat Rev Drug Discov 2003; 2: 214–221.
Terregino CA et al. Endogenous mediators in emergency department patients with presumed sepsis: are levels associated with progression to severe sepsis and death? Ann Emerg Med 2000; 35: 26–34.
Wolins N et al. Intravenous administration of replication-incompetent adenovirus to rhesus monkeys induces thrombocytopenia by increasing in vivo platelet clearance. Br J Hematol 2003; 123: 903–905.
Zucker-Franklin D . Platelets in viral infection. In: Joseph M (ed). Immunopharmacology of Platelets. Elsevier Science & Technology: London, 1995, pp 137–151.
Davis FF . The origin of pegnology. Adv Drug Deliv Rev 2002; 54: 457–458.
Working PK, Newman MS, Johnson J, Cornacoff JB . Safety of poly(ethylene glycol) and poly(ethylene glycol) derivatives. In: Harris JM, Zalipsky S (eds). Poly(ethylene glycol) Chemistry and Biological Applications. American Chemical Society: Washington, DC, 1997, pp 45–57.
Zou L, Zhou H, Pastore L, Yang K . Prolonged transgene expression mediated by a helper-dependent adenoviral vector (hdAd) in the central nervous system. Mol Ther 2000; 2: 105–113.
Schiedner G et al. Genomic DNA transfer with a high-capacity adenovirus vector results in improved in vivo gene expression and decreased toxicity. Nat Genet 1998; 18: 180–183.
Parks RJ et al. A helper-dependent adenovirus vector system: removal of helper virus by cre-mediated excision of the viral packaging signal. Proc Natl Acad Sci USA 1996; 93: 13565–13570.
Zhou H, Pastore L, Beaudet AL . Helper-dependent adenoviral vectors. Meth Enzymol 2002; 346: 177–198.
Maizel Jr JV, White DO, Scharff MD . The polypeptides of adenovirus. I. Evidence for multiple protein components in the virion and a comparison of types 2, 7A, and 12. Virology 1968; 36: 115–125.
O'Riordan C et al. PEGylation of adenovirus with retention of infectivity and protection from neutralizing antibody in vitro and in vivo. Hum Gene Ther 1999; 10: 1349–1358.
Acknowledgements
We would like to thank Shellie Callahan for invaluable technical assistance with animal handling and vector administration. This work was funded by a New Investigator grant from the American Foundation for Pharmaceutical Education and the Burroughs Wellcome Fund (MAC).
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Croyle, M., Le, H., Linse, K. et al. PEGylated helper-dependent adenoviral vectors: highly efficient vectors with an enhanced safety profile. Gene Ther 12, 579–587 (2005). https://doi.org/10.1038/sj.gt.3302441
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DOI: https://doi.org/10.1038/sj.gt.3302441
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