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The Fate of Plasmid DNA After Intravenous Injection in Mice: Involvement of Scavenger Receptors in Its Hepatic Uptake

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Abstract

Purpose. We examined the stability and disposition characteristics of a naked plasmid DNA pCAT as a model gene after intravenous injection in mice to construct the strategy of in vivo gene delivery systems.

Methods. After the injection of pCAT to the mice, stability, tissue distribution, hepatic cellular localization, and effect of some polyanions on the hepatic uptake were studied.

Results. The in vitro study demonstrated that the pCAT was rapidly degraded in mouse whole blood with a half-life of approximately 10 min at a concentration of 100 µg/ml. After intravenous injection, pCAT was degraded at a significantly faster rate than that observed in the whole blood, suggesting that pCAT in vivo was also degraded in other compartments. Following intravenous injection of [32P] pCAT, radioactivity was rapidly eliminated from the plasma due to extensive uptake by the liver. Hepatic accumulation occurred preferentially in the non-parenchymal cells. The hepatic uptake of radioactivity derived from [32P] pCAT was inhibited by preceding administration of polyanions such as polyinosinic acid, dextran sulfate, maleylated and succinylated bovine serum albumin but not by polycytidylic acid. These findings indicate that pCAT is taken up by the liver via scavenger receptors on the non-parenchymal cells. Pharmacokinetic analysis revealed that the apparent hepatic uptake clearance was fairly close to the liver plasma flow.

Conclusions. These findings provide useful information for the development of delivery systems for in vivo gene therapy.

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REFERENCES

  1. R. C. Mulligan. The basic science of gene therapy. Science 260:926–932 (1993)

    Google Scholar 

  2. S. J. Szilvassy, C. C. Fraser, C. J. Eaves, P. M. Lansdorp, A. C. Eaves, and R. K. Humphries. Retrovirus-mediated gene transfer to purified hemopoietic stem cells with long-term lympho-myelopoietic repopulating ability. Proc. Natl. Acad. Sci. USA. 86:8798–8802 (1989)

    Google Scholar 

  3. M. A. Rosenfeld, K. Yoshimura, B. C. Trapnell, K. Yoneyama, E. R. Rosenthal, W. Dalemans, M. Fukayama, J. Bargon, A. Stier, J-P. Lecocq, and R. G. Crystal. In vivo transfer of the human cystic fibrosis transmembrane conductance regulator gene to the airway epithelium. Cell 68:143–155 (1992)

    Google Scholar 

  4. G. Y. Wu and C. H. Wu. Receptor-mediated gene delivery and expression in vivo. J. Biol. Chem. 263:14621–14624 (1988)

    Google Scholar 

  5. C. H. Wu, J. M. Wilson, and G. Y. Wu. Targeting genes: Delivery and persistent expression of a foreign gene driven by mammalian regulatory elements in vivo. J. Biol. Chem. 264:16985–16987 (1989)

    Google Scholar 

  6. J. C. Perales, T. Ferkol, H. Beegen, O. D. Ratnoff, and R. W. Hanson. Gene transfer in vivo: Sustained expression and regulation of genes introduced into the liver by receptor-targeted uptake. Proc. Natl. Acad. Sci. USA. 91:4086–4090 (1994)

    Google Scholar 

  7. D. T. Curiel, S. Agrawal, E. Wagner, and M. Cotten. Adenovirus enhancement of transferrin-polylysine-mediated gene delivery. Proc. Natl. Acad. Sci. USA. 88:8850–8854 (1991)

    Google Scholar 

  8. P. L. Feigner and G. M. Ringold. Cationic liposome-mediated transfection. Nature 337:387–388 (1989)

    Google Scholar 

  9. J. G. Smith, R. L. Walzem, and J. B. German. Liposomes as agents of DNA transfer. Biochim. Biophys. Acta. 1154:327–340 (1993)

    Google Scholar 

  10. N. Zhu, D. Liggitt, Y. Liu, and R. Debs. Systemic gene expression after intravenous DNA delivery into adult mice. Science 261:209–211 (1993)

    Google Scholar 

  11. H. Imoto, Y. Sakamura, K. Ohkouchi, R. Atsumi, Y. Takakura, H. Sezaki, and M. Hashida. Disposition Characteristics of macromolecules in the perfused tissue-isolated tumor preparation. Cancer Res. 52:4396–4401 (1992)

    Google Scholar 

  12. T. Fujita, M. Nishikawa, C. Tamaki, Y. Takakura, M. Hashida, and H. Sezaki. Targeted delivery of human recombinant superoxide dismutase by chemical modification with mono-and polysaccharide derivatives. J. Pharmacol. Exp. Ther. 263:971–978 (1992)

    Google Scholar 

  13. T. Fujita, H. Furitsu, M. Nishikawa, Y. Takakura, H. Sezaki, and M. Hashida. Therapeutic effects of superoxide dismutase derivatives modified with mono-and polysaccharide on hepatic injury induced by ischemia/reperfusion. Biochem. Biophys. Res. Commun. 189:191–196 (1992)

    Google Scholar 

  14. Y. Takakura, S. Masuda, H. Tokuda, M. Nishikawa, and M. Hashida. Targeted delivery of superoxide dismutase to macrophages via mannose receptor-mediated mechanism. Biochem. Pharmacol. 47:853–858 (1994)

    Google Scholar 

  15. Y. Takakura, T. Fujita, H. Furitsu, M. Nishikawa, H. Sezaki, and M. Hashida. Pharmacokinetics of succinylated proteins and dextran sulfate in mice: Implications for hepatic targeting of protein drugs by direct succinylation via scavenger receptors. Int. J. Pharm. 105:19–29 (1994)

    Google Scholar 

  16. T. Miyao, Y. Takakura, and M. Hashida. Stability and in vivo disposition characteristics of oligonucleotides and oligonucleotide conjugated with macromolecule. Proc. of the 20th Symp. on Controlled Release of Bioactive Materials, Washington, D.C., USA. 492–493 (1993)

  17. J. Sambrook, E. F. Fritsch, and T. Maniatis. Molecular Cloning: A Laboratory Manual (Cold Spring Harbor Lab. Press, Plainview, NY), 2nd Ed. (1989)

    Google Scholar 

  18. Y. Takakura, T. Fujita, M. Hashida, and H. Sezaki. Disposition characteristics of macromolecules in tumor-bearing mice. Pharm. Res. 7:339–345 (1990)

    Google Scholar 

  19. S. Nakane, S. Matsumoto, Y. Takakura, M. Hashida, and H. Sezaki. The accumulation mechanism of cationic mitomycin C-dextran conjugates in the liver: In-vivo cellular localization and in-vitro interaction with hepatocytes. J. Pharm. Pharmacol. 40:1–6 (1988)

    Google Scholar 

  20. K. Yamaoka, Y. Tanigawara, H. Tanaka, and Y. Uno. A pharmacokinetic analysis program (MULTI) for microcomputer. J. Pharmacobio-Dyn. 4:879–885 (1981)

    Google Scholar 

  21. M. Nishikawa, Y. Ohtsubo, J. Ohno, T. Fujita, Y. Koyama, F. Yamashita, M. Hashida, and H. Sezaki. Pharmacokinetics of receptor-mediated hepatic uptake of glycosylated albumin in mice. Int. J. Pharm. 85:75–85 (1993)

    Google Scholar 

  22. Ch. Gosse, J. B. Le Pecq, P. Defrance, and C. Paoletti. Initial degradation of deoxyribonucleic acid after injection in mammals. Cancer Res. 25:877–883 (1965)

    Google Scholar 

  23. W. Emlen, A. Rifai, D. Magilavy, and M. Mannik. Hepatic binding of DNA is mediated by a receptor on nonparenchymal cells. Am. J. Pathol. 133:54–60 (1988)

    Google Scholar 

  24. T. Tsumita and M. Iwanaga. Fate of injected deoxyribonucleic acid in mice. Nature 198, 1088–1089 (1963)

    Google Scholar 

  25. W. Emlen and M. Mannik. Kinetics and mechanisms for removal of circulating single-stranded DNA in mice. J. Exp. Med. 147:684–699 (1978)

    Google Scholar 

  26. W. Emlen and M. Mannik. Effect of DNA size and strandedness on the in vivo clearance and organ localization of DNA. Clin. Exp. Immunol. 56:185–192 (1984).

    Google Scholar 

  27. F. G. Cosio, L. A. Hebert, D. J. Birmingham, B. L. Dorval, A. P. Bakaletz, G. A. Kujala, J. C. Edberg, and R. P. Taylor. Clearance of human antibody/DNA immune complexes and free DNA from the circulation of the nonhuman primate. Clin. Immunol. Immunopathol. 42:1–9 (1987)

    Google Scholar 

  28. “Handbook of Physiology,” W. F. Hamilton and P. Dow. Eds., American Physiological Society, Washington, D.C., sect. 2. (1962)

    Google Scholar 

  29. M. Krieger, S. Acton, J. Ashkenas, A. Pearson, M. Penman, and D. Resnick. Molecular flypaper, host defense, and atherosclerosis: Structure, binding properties and functions of macrophage scavenger receptors. J. Biol. Chem. 268:4569–4572 (1993)

    Google Scholar 

  30. Y. B. De Rijke and J. C. Van Berkel. Rat liver Kupffer and endothelial cells express different binding proteins for modified low density lipoproteins. J. Biol. Chem. 269:824–827 (1994)

    Google Scholar 

  31. T. Kodama, M. Freeman, L. Rohrer, J. Zabrecky, P. Matsudaira, and M. Krieger. Type I macrophage scavenger receptor contains α-helical and collagen-like coiled coils. Nature 343, 531–535 (1990)

    Google Scholar 

  32. L. Rohrer, M. Freeman, T. Kodama, M. Penman, and M. Krieger. Coiled-coil fibrous domains mediate ligand binding by macrophage scavenger receptor type II. Nature 343:570–572 (1990)

    Google Scholar 

  33. S. Acton, D. Resnick, M. Freeman, Y. Ekkel, J. Ashkenas, and M. Krieger. The collagenous domains of macrophage scavenger receptors and complement component Clq mediate their similar, but not identical, binding specificities for polyanionic ligands. J. Biol. Chem. 268:3530–3537 (1993)

    Google Scholar 

  34. M. R. Van Schravendijk and R. A. Dwek. Interaction of C1q with DNA. Mol. Immunol. 19:1179–1187 (1982).

    Google Scholar 

  35. A. M. Pearson, A. Rich, and M. Krieger. Polynucleotide binding to macrophage scavenger receptors depends on the formation of base-quartet-stabilized four-stranded helices. J. Biol. Chem. 268:3546–3554 (1993)

    Google Scholar 

  36. J. M. Harris, ed. Poly (Ethylene Glycol) Chemistry: Biotechnical and Biomedical Applications. New York: Plenum Press (1992)

    Google Scholar 

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Authors and Affiliations

  1. Department of Drug Delivery Research, Faculty of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto, 606-01, Japan

    Kenji Kawabata, Yoshinobu Takakura & Mitsuru Hashida

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  1. Kenji Kawabata
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  2. Yoshinobu Takakura
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  3. Mitsuru Hashida
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Kawabata, K., Takakura, Y. & Hashida, M. The Fate of Plasmid DNA After Intravenous Injection in Mice: Involvement of Scavenger Receptors in Its Hepatic Uptake. Pharm Res 12, 825–830 (1995). https://doi.org/10.1023/A:1016248701505

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