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. 1993 Feb 25;21(4):871–878. doi: 10.1093/nar/21.4.871

Specific gene transfer mediated by lactosylated poly-L-lysine into hepatoma cells.

P Midoux 1, C Mendes 1, A Legrand 1, J Raimond 1, R Mayer 1, M Monsigny 1, A C Roche 1
PMCID: PMC309219  PMID: 8383843

Abstract

Plasmid DNA/glycosylated polylysine complexes were used to transfer in vitro a luciferase reporter gene into human hepatoma cells by a receptor-mediated endocytosis process. HepG2 cells which express a galactose specific membrane lectin were efficiently and selectively transfected with pSV2Luc/lactosylated polylysine complexes in a sugar dependent manner: i) HepG2 cells which do not express membrane lectin specific for mannose were quite poorly transfected with pSV2Luc/mannosylated polylysine complexes, ii) HeLa cells which do not express membrane lectin specific for galactose were not transfected with pSV2Luc/lactosylated polylysine complexes. The transfection efficiency of HepG2 cells with pSV2Luc/lactosylated polylysine complexes was greatly enhanced either in the presence of chloroquine or in the presence of a fusogenic peptide. A 22-residue peptide derived from the influenza virus hemagglutinin HA2 N-terminal polypeptide that mimics the fusogenic activity of the virus, was selected. In the presence of the fusogenic peptide, the luciferase activity in HepG2 cells was 10 fold larger than that of cells transfected with pSV2Luc/lactosylated polylysine complexes in the presence of chloroquine.

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Selected References

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  1. Ashwell G., Harford J. Carbohydrate-specific receptors of the liver. Annu Rev Biochem. 1982;51:531–554. doi: 10.1146/annurev.bi.51.070182.002531. [DOI] [PubMed] [Google Scholar]
  2. Bonfils E., Depierreux C., Midoux P., Thuong N. T., Monsigny M., Roche A. C. Drug targeting: synthesis and endocytosis of oligonucleotide-neoglycoprotein conjugates. Nucleic Acids Res. 1992 Sep 11;20(17):4621–4629. doi: 10.1093/nar/20.17.4621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bonfils E., Mendes C., Roche A. C., Monsigny M., Midoux P. Uptake by macrophages of a biotinylated oligo-alpha-deoxythymidylate by using mannosylated streptavidin. Bioconjug Chem. 1992 Jul-Aug;3(4):277–284. doi: 10.1021/bc00016a004. [DOI] [PubMed] [Google Scholar]
  4. Brandley B. K., Swiedler S. J., Robbins P. W. Carbohydrate ligands of the LEC cell adhesion molecules. Cell. 1990 Nov 30;63(5):861–863. doi: 10.1016/0092-8674(90)90487-y. [DOI] [PubMed] [Google Scholar]
  5. Cotten M., Längle-Rouault F., Kirlappos H., Wagner E., Mechtler K., Zenke M., Beug H., Birnstiel M. L. Transferrin-polycation-mediated introduction of DNA into human leukemic cells: stimulation by agents that affect the survival of transfected DNA or modulate transferrin receptor levels. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4033–4037. doi: 10.1073/pnas.87.11.4033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cotten M., Wagner E., Zatloukal K., Phillips S., Curiel D. T., Birnstiel M. L. High-efficiency receptor-mediated delivery of small and large (48 kilobase gene constructs using the endosome-disruption activity of defective or chemically inactivated adenovirus particles. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6094–6098. doi: 10.1073/pnas.89.13.6094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Curiel D. T., Agarwal S., Wagner E., Cotten M. Adenovirus enhancement of transferrin-polylysine-mediated gene delivery. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8850–8854. doi: 10.1073/pnas.88.19.8850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Curiel D. T., Wagner E., Cotten M., Birnstiel M. L., Agarwal S., Li C. M., Loechel S., Hu P. C. High-efficiency gene transfer mediated by adenovirus coupled to DNA-polylysine complexes. Hum Gene Ther. 1992 Apr;3(2):147–154. doi: 10.1089/hum.1992.3.2-147. [DOI] [PubMed] [Google Scholar]
  9. Derrien D., Midoux P., Petit C., Nègre E., Mayer R., Monsigny M., Roche A. C. Muramyl dipeptide bound to poly-L-lysine substituted with mannose and gluconoyl residues as macrophage activators. Glycoconj J. 1989;6(2):241–255. doi: 10.1007/BF01050652. [DOI] [PubMed] [Google Scholar]
  10. Gruenberg J., Howell K. E. Membrane traffic in endocytosis: insights from cell-free assays. Annu Rev Cell Biol. 1989;5:453–481. doi: 10.1146/annurev.cb.05.110189.002321. [DOI] [PubMed] [Google Scholar]
  11. Kaiser E., Colescott R. L., Bossinger C. D., Cook P. I. Color test for detection of free terminal amino groups in the solid-phase synthesis of peptides. Anal Biochem. 1970 Apr;34(2):595–598. doi: 10.1016/0003-2697(70)90146-6. [DOI] [PubMed] [Google Scholar]
  12. Kielian M., Jungerwirth S. Mechanisms of enveloped virus entry into cells. Mol Biol Med. 1990 Feb;7(1):17–31. [PubMed] [Google Scholar]
  13. Knowles B. B., Howe C. C., Aden D. P. Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen. Science. 1980 Jul 25;209(4455):497–499. doi: 10.1126/science.6248960. [DOI] [PubMed] [Google Scholar]
  14. Lear J. D., DeGrado W. F. Membrane binding and conformational properties of peptides representing the NH2 terminus of influenza HA-2. J Biol Chem. 1987 May 15;262(14):6500–6505. [PubMed] [Google Scholar]
  15. Maxfield F. R. Weak bases and ionophores rapidly and reversibly raise the pH of endocytic vesicles in cultured mouse fibroblasts. J Cell Biol. 1982 Nov;95(2 Pt 1):676–681. doi: 10.1083/jcb.95.2.676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mellman I., Fuchs R., Helenius A. Acidification of the endocytic and exocytic pathways. Annu Rev Biochem. 1986;55:663–700. doi: 10.1146/annurev.bi.55.070186.003311. [DOI] [PubMed] [Google Scholar]
  17. Midoux P., Roche A. C., Monsigny M. Quantitation of the binding, uptake, and degradation of fluoresceinylated neoglycoproteins by flow cytometry. Cytometry. 1987 May;8(3):327–334. doi: 10.1002/cyto.990080314. [DOI] [PubMed] [Google Scholar]
  18. Monsigny M., Petit C., Roche A. C. Colorimetric determination of neutral sugars by a resorcinol sulfuric acid micromethod. Anal Biochem. 1988 Dec;175(2):525–530. doi: 10.1016/0003-2697(88)90578-7. [DOI] [PubMed] [Google Scholar]
  19. Monsigny M., Roche A. C., Kieda C., Midoux P., Obrénovitch A. Characterization and biological implications of membrane lectins in tumor, lymphoid and myeloid cells. Biochimie. 1988 Nov;70(11):1633–1649. doi: 10.1016/0300-9084(88)90299-4. [DOI] [PubMed] [Google Scholar]
  20. Monsigny M., Roche A. C., Midoux P. Uptake of neoglycoproteins via membrane lectin(s) of L1210 cells evidenced by quantitative flow cytofluorometry and drug targeting. Biol Cell. 1984;51(2):187–196. doi: 10.1111/j.1768-322x.1984.tb00298.x. [DOI] [PubMed] [Google Scholar]
  21. Murata M., Kagiwada S., Hishida R., Ishiguro R., Ohnishi S., Takahashi S. Modification of the N-terminus of membrane fusion-active peptides blocks the fusion activity. Biochem Biophys Res Commun. 1991 Sep 16;179(2):1050–1055. doi: 10.1016/0006-291x(91)91925-3. [DOI] [PubMed] [Google Scholar]
  22. Murata M., Kagiwada S., Takahashi S., Ohnishi S. Membrane fusion induced by mutual interaction of the two charge-reversed amphiphilic peptides at neutral pH. J Biol Chem. 1991 Aug 5;266(22):14353–14358. [PubMed] [Google Scholar]
  23. Murata M., Sugahara Y., Takahashi S., Ohnishi S. pH-dependent membrane fusion activity of a synthetic twenty amino acid peptide with the same sequence as that of the hydrophobic segment of influenza virus hemagglutinin. J Biochem. 1987 Oct;102(4):957–962. doi: 10.1093/oxfordjournals.jbchem.a122137. [DOI] [PubMed] [Google Scholar]
  24. Murata M., Takahashi S., Kagiwada S., Suzuki A., Ohnishi S. pH-dependent membrane fusion and vesiculation of phospholipid large unilamellar vesicles induced by amphiphilic anionic and cationic peptides. Biochemistry. 1992 Feb 25;31(7):1986–1992. doi: 10.1021/bi00122a013. [DOI] [PubMed] [Google Scholar]
  25. Roche A. C., Barzilay M., Midoux P., Junqua S., Sharon N., Monsigny M. Sugar-specific endocytosis of glycoproteins by Lewis lung carcinoma cells. J Cell Biochem. 1983;22(3):131–140. doi: 10.1002/jcb.240220302. [DOI] [PubMed] [Google Scholar]
  26. Roche A. C., Midoux P., Pimpaneau V., Nègre E., Mayer R., Monsigny M. Endocytosis mediated by monocyte and macrophage membrane lectins--application to antiviral drug targeting. Res Virol. 1990 Mar-Apr;141(2):243–249. doi: 10.1016/0923-2516(90)90028-h. [DOI] [PubMed] [Google Scholar]
  27. Rosenkranz A. A., Yachmenev S. V., Jans D. A., Serebryakova N. V., Murav'ev V. I., Peters R., Sobolev A. S. Receptor-mediated endocytosis and nuclear transport of a transfecting DNA construct. Exp Cell Res. 1992 Apr;199(2):323–329. doi: 10.1016/0014-4827(92)90441-a. [DOI] [PubMed] [Google Scholar]
  28. Schwartz A. L., Fridovich S. E., Knowles B. B., Lodish H. F. Characterization of the asialoglycoprotein receptor in a continuous hepatoma line. J Biol Chem. 1981 Sep 10;256(17):8878–8881. [PubMed] [Google Scholar]
  29. Sharon N., Lis H. Lectins as cell recognition molecules. Science. 1989 Oct 13;246(4927):227–234. doi: 10.1126/science.2552581. [DOI] [PubMed] [Google Scholar]
  30. Smythe E., Warren G. The mechanism of receptor-mediated endocytosis. Eur J Biochem. 1991 Dec 18;202(3):689–699. doi: 10.1111/j.1432-1033.1991.tb16424.x. [DOI] [PubMed] [Google Scholar]
  31. Takahashi S. Conformation of membrane fusion-active 20-residue peptides with or without lipid bilayers. Implication of alpha-helix formation for membrane fusion. Biochemistry. 1990 Jul 3;29(26):6257–6264. doi: 10.1021/bi00478a021. [DOI] [PubMed] [Google Scholar]
  32. Wagner E., Cotten M., Foisner R., Birnstiel M. L. Transferrin-polycation-DNA complexes: the effect of polycations on the structure of the complex and DNA delivery to cells. Proc Natl Acad Sci U S A. 1991 May 15;88(10):4255–4259. doi: 10.1073/pnas.88.10.4255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wagner E., Cotten M., Mechtler K., Kirlappos H., Birnstiel M. L. DNA-binding transferrin conjugates as functional gene-delivery agents: synthesis by linkage of polylysine or ethidium homodimer to the transferrin carbohydrate moiety. Bioconjug Chem. 1991 Jul-Aug;2(4):226–231. doi: 10.1021/bc00010a006. [DOI] [PubMed] [Google Scholar]
  34. Wagner E., Zatloukal K., Cotten M., Kirlappos H., Mechtler K., Curiel D. T., Birnstiel M. L. Coupling of adenovirus to transferrin-polylysine/DNA complexes greatly enhances receptor-mediated gene delivery and expression of transfected genes. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6099–6103. doi: 10.1073/pnas.89.13.6099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wagner E., Zatloukal K., Cotten M., Kirlappos H., Mechtler K., Curiel D. T., Birnstiel M. L. Coupling of adenovirus to transferrin-polylysine/DNA complexes greatly enhances receptor-mediated gene delivery and expression of transfected genes. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6099–6103. doi: 10.1073/pnas.89.13.6099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wagner E., Zenke M., Cotten M., Beug H., Birnstiel M. L. Transferrin-polycation conjugates as carriers for DNA uptake into cells. Proc Natl Acad Sci U S A. 1990 May;87(9):3410–3414. doi: 10.1073/pnas.87.9.3410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Weiss S. J., Test S. T., Eckmann C. M., Roos D., Regiani S. Brominating oxidants generated by human eosinophils. Science. 1986 Oct 10;234(4773):200–203. doi: 10.1126/science.3018933. [DOI] [PubMed] [Google Scholar]
  38. White J. M. Viral and cellular membrane fusion proteins. Annu Rev Physiol. 1990;52:675–697. doi: 10.1146/annurev.ph.52.030190.003331. [DOI] [PubMed] [Google Scholar]
  39. White J., Kielian M., Helenius A. Membrane fusion proteins of enveloped animal viruses. Q Rev Biophys. 1983 May;16(2):151–195. doi: 10.1017/s0033583500005072. [DOI] [PubMed] [Google Scholar]
  40. Wilson J. M., Grossman M., Wu C. H., Chowdhury N. R., Wu G. Y., Chowdhury J. R. Hepatocyte-directed gene transfer in vivo leads to transient improvement of hypercholesterolemia in low density lipoprotein receptor-deficient rabbits. J Biol Chem. 1992 Jan 15;267(2):963–967. [PubMed] [Google Scholar]
  41. Wu C. H., Wilson J. M., Wu G. Y. Targeting genes: delivery and persistent expression of a foreign gene driven by mammalian regulatory elements in vivo. J Biol Chem. 1989 Oct 15;264(29):16985–16987. [PubMed] [Google Scholar]
  42. Wu G. Y., Wilson J. M., Shalaby F., Grossman M., Shafritz D. A., Wu C. H. Receptor-mediated gene delivery in vivo. Partial correction of genetic analbuminemia in Nagase rats. J Biol Chem. 1991 Aug 5;266(22):14338–14342. [PubMed] [Google Scholar]
  43. Wu G. Y., Wu C. H. Evidence for targeted gene delivery to Hep G2 hepatoma cells in vitro. Biochemistry. 1988 Feb 9;27(3):887–892. doi: 10.1021/bi00403a008. [DOI] [PubMed] [Google Scholar]
  44. Wu G. Y., Wu C. H. Receptor-mediated gene delivery and expression in vivo. J Biol Chem. 1988 Oct 15;263(29):14621–14624. [PubMed] [Google Scholar]
  45. Wu G. Y., Wu C. H. Receptor-mediated in vitro gene transformation by a soluble DNA carrier system. J Biol Chem. 1987 Apr 5;262(10):4429–4432. [PubMed] [Google Scholar]
  46. Zenke M., Steinlein P., Wagner E., Cotten M., Beug H., Birnstiel M. L. Receptor-mediated endocytosis of transferrin-polycation conjugates: an efficient way to introduce DNA into hematopoietic cells. Proc Natl Acad Sci U S A. 1990 May;87(10):3655–3659. doi: 10.1073/pnas.87.10.3655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. de Wet J. R., Wood K. V., DeLuca M., Helinski D. R., Subramani S. Firefly luciferase gene: structure and expression in mammalian cells. Mol Cell Biol. 1987 Feb;7(2):725–737. doi: 10.1128/mcb.7.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]

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