Delivery systems for gene therapy
Cite this article as: Wu, G.Y. & Wu, C.H. Biotherapy (1991) 3: 87. doi:10.1007/BF02175102 Abstract
Introduction of foreign genes into mammalian cells
in vitro has been accomplished previously by a variety of methods. The few techniques that have been developed for transfection of mammalian cells in vivo, are technically difficult or lack cell specificity.
We have developed a soluble, targetable DNA carrier system consisting of an asialoglycoprotein covalently coupled to a polycation. The strategy was based on: 1) the presence of unique receptors on hepatocytes which internalize galactose-terminal (asialo-)glycoproteins; 2) polycations can bind DNA in a non-covalent, non-damaging interaction. Using chloramphenicol acetyltransferase (CAT) as a marker gene, specific delivery and expression of CAT was demonstrated
in vitro using asialoglycoprotein receptor ( +) and (-) cell lines.
Intravenous injection of conjugate-DNA complexes in rats resulted in detection of CAT DNA sequences in liver 10 min later by dot blots with a CAT cDNA probe. CAT enzyme activity 24 hrs later was found specifically in liver but no other tissues or control livers. Targeted hepatic CAT expression was transient, maximal at 24 hrs but declined to barely detectable levels by 96 hrs. Persistent foreign gene expression was achieved by injection of DNA complex followed by 67% partial hepatectomy. High levels of hepatic CAT activity were detected through 11 weeks post-hepatectomy.
The data indicate that a targetable gene delivery system can permit
in vivo expression of an exogenous gene after simple intravenous injection. The foreign gene expression can be enhanced and made to persist by induction of hepatocyte replication. Key words receptor-mediated endocytosis targeting genes hepatocytes asialoglycoproteins References
Graham FL, Van der Eb AJ. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 1973; 52: 456–67.
Gopal TV. Gene transfer method for transient gene expression, stable transformation, and co-transformation of suspension cell cultures. Mole Cell Biol 1985; 5: 1183–93.
Potter H, Weir L, Leder P. Enhancer-dependent expression of human k immunoglobulin genes introduced into mouse pre-B lymphocytes by electroporation. Proc Natl Acad Sci 1984; 81: 7161–5.
Harland R, Weintraub H. Translation of mRNA injected into Xenopus oocytes is specifically inhibited by antisense RNA. J Cell Biol 1985; 101: 1094–9.
Fechheimer M, Boylan JF, Parker S, Sisken JE, Patel GL, Zimmer SG. Transfection of mammalian cells with plasmid DNA by scrape-loading and sonication loading. Proc Natl Acad Sci 1987; 84: 8463–7.
Nicolau C, Sene C. Liposome-mediated DNA transfer in eukaryotic cells. Biochim Biophys Acta 1982; 721: 185–90.
Fraley RT, Fornari CS, Kaplan S. Entrapment of a bacterial plasmid in phospholipid vesicles: Potential for gene transfer. Proc Natl Acad Sci USA 1979; 76: 3348–52.
Guild BC, Finer MH, Housman DE, Mulligan RC. Development of retrovirus vectors useful for expressing genes in cultured murine embryonal cells and hematopoietic cells
. J Virology 1988; 62: 3795–801.
Zwiebel JA, Freeman SM, Kantoff PW, Cornetta K, Ryan US, Anderson WF. High level recombinant gene expression in rabbit endothelial cells transduced by retro-viral vectors. Science 1989; 243: 220–2.
Dubensky TW, Campbell BA, Villarreal LP. Direct transfection of viral and plasmid DNA into liver or spleen of mice. Proc Natl Acad Sci USA 1984; 81: 7529–33.
Benvenisty N, Reshef L. Direct introduction of genes into rats and expression of the genes. Proc Natl Acad Sci USA 1986; 83: 9551–5.
Nicolau C, Le Pape A, Soriano P, Fargette F, Juhel M-F.
expression of rat insulin after intravenous administration of the liposome-entrapped gene for rat insulin 1. Proc Natl Acad Sci USA 1983; 80: 1068–72.
Wang, C-Y, Huang L. pH-sensitive immunoliposomes mediate target-cell specific delivery and controlled expression of a foreign gene in mouse. Proc Natl Acad Sci USA 1987; 84: 7851–5.
Soriano P, Dijkstra J, Legrand A, Spanjer H, Londos-Gagliardi D, Roerdink F, Scherphof G, Nicolau C. Targeted and nontargeted liposomes for
transfer to rat liver cells of a plasmid containing the preproinsulin I gene. Proc Natl Acad Sci USA 1983; 80: 7128–31.
Kucherlapati R, Skoultchi AI. Introduction of purified genes into mammalian cells. In: CRC critical reviews in biochemistry, Boca Raton, FL: Chemical Rubber Company 1984; 349–79.
Eglitis MA, Kantoff PW, Jolly JD, Jones JB, Anderson WF, Lothrup CD Jr. Gene transfer into hemotopoietic cells from normal and cyclic hematopoietic dogs using retroviral vectors. Blood 1988; 71: 717–22.
Kantoff PW, Freeman SM, Anderson WF. Prospects for gene therapy for immunodeficiency diseases. Ann Rev Immunol 1988; 6: 591–4.
Wilson JM, Jefferson DM, Roy Chowdhury J, Novikoff PM, Johnston DE, Mulligan RC. Retrovirus-mediated transduction of adult hepatocytes. Proc Natl Acad Sci USA 1988; 85: 3014–8.
Wilson JM, Johnston DE, Jefferson DM, Mulligan RC. Correction of the genetic defect in hepatocytes from the Watenabe heritable hyperlipidemic rabbit. Proc Natl Acad Sci USA 1988; 85: 4421–5.
Ashwell C, Morell AG. The role of surface carbohydrates in the hepatic recognition and transport of circulating glycoproteins. Adv Enzymol 1974; 41: 99–128.
Wall DA, Wilson G, Hubbard AL. The galactose specific recognition system of mammalian liver: Receptor distribution on the hepatocyte surface. Cell 1980; 21: 79–93.
Dunn WA, Hubbard AL, Aronson NN. Low temperature selectively inhibits fusion of pinocytic vesicles and lysosomes during heterophagy of
I-asialofetuin by the perfused rat liver. J Biol Chem 1980; 255: 5971–8.
Wu GY, Wu CH, Stockert RJ. A model for the specific rescue of normal hepatocytes during methotrexate treatment of hepatic malignancy. Proc Natl Acad Sci USA 1983; 80: 3078–80.
Wu GY, Wu CH, Rubin MI. Acetaminophen hepatotoxicity and targeted rescue: A model for specific chemotherapy for hepatocellular carcinoma. Hepatology 1985; 5: 709–13.
Wu GY, Keegan-Rogers V, Franklin S, Midford S, Wu CH. Targeted antagonism of galactosamine toxicity in normal rat hepatocytes
. J Biol Chem 1988; 263: 4719–23.
Jung G, Kohnlein W, Luders G. Biological activity of the antitumor protein neocarzinostatin coupled to a monoclonal antibody by N-succinimidyl 3-(2-pyridylthio)-propionate. Biochem Biophys Res Commun 1981; 101: 599–606.
Laemmli UK. Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature (Lond) 1970; 227: 680–5.
Wu GY, Wu CH. Receptor-mediated in vitro gene transformation by a soluble DNA carrier system. J Biol Chem 1987; 262: 4429–32.
Wu GY, Wu CH. Evidence for targeted gene delivery to Hep G2 hepatoma cells
. Biochemistry 1988; 27: 887–92.
Selden RF, Howie KB, Rowe ME, Goodman HM, Moore DD. Human growth hormone as a reporter gene in regulation studies employing transient gene expression. Mol Cell Biol 1986; 6: 3173–9.
Wu GY, Wu CH. Receptor-mediated gene delivery and expression
. J Biol Chem 1988; 263: 14621–4.
Varmus HE, Padgett T, Heasley S, Simon G, Bishop JM. Cellular functions are required for the synthesis and integration of avian sarcoma virus-specific DNA. Cell 1977; 11–307.
Leffert HL, Koch KS, Moran T, Rubalcava B. Hormonal control of rat liver regeneration. Gastroenterology 1979; 76: 1470–82.
Wu GY, Wilson JM, Wu CH. Targeting genes: Delivery and persistent expression of a foreign gene driven by mammalian regulatory elements
in vivo. J Biol Chem 1989; 254: 16985–7. Copyright information
© Kluwer Academic Publishers 1991