Biopolym. Cell. 2004; 20(1-2):25-32.
Non-viral gene delivery of human apoA1 into mammalian cells in vitro and in vivo
- Institute of Molecular Biology and Genetics, NAS of Ukraine
150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
- Gerontology Institute of the Academy of Medical Sciences of Ukraine
67, Vyshgorodska Str., Kyiv, Ukraine, 04114
Some approaches to the gene therapy of atherosclerosis by non-viral transfer of human apoA1 gene, encoding the main protein component of antiatherogenic lipoproteins, into liver cells are being studied. The plasmid vector for the expression of the human genomic apoA1 gene in mammalian cells has been constructed. The construction contains bacterial plasmid sequences for its replication in Escherihia coli and expression cassette for human apoA1 gene flanked by inverted terminal repeats of human adeno-associated virus. The cationic polymer polyethylenimine of 25 kD (PEI) has been used for gene delivery. It has been shown, that 50–70 % of cells express transgene after PEI-mediated in vitro transfection with analogical vector containing reporter green fluorescence protein gene. The experiments on animals have demonstrated, that the human protein ApoA1 is being tested in animal plasma during 15 days after apoA1 gene injection in the liver. PCR-analysis of total liver DNA of experimental animal has shown the presence of expected fragment of apoA1 gene in rat liver in 15 days after the gene delivery.
 Strauss M., Barranger J.A. Concepts In Gene Therapy, Berlin: DeGruyter 1997 p. 553.
 Xanthopoutos, K.G. Gene Therapy. NATO ASI Series: H: Cell Biology, Berlin; Heidelberg: Springer 1998 p. 238.
 Chong PH, Bachenheimer BS. Current, new and future treatments in dyslipidaemia and atherosclerosis. Drugs. 2000;60(1):55-93.
 O'Brien T, Simari RD. Gene therapy for atherosclerotic cardiovascular disease: a time for optimism and caution. Mayo Clin Proc. 2000;75(8):831-4.
 Sinnaeve P, Varenne O, Collen D, Janssens S. Gene therapy in the cardiovascular system: an update. Cardiovasc Res. 1999;44(3):498-506.
 Belalcazar M, Chan L. Somatic gene therapy for dyslipidemias. J Lab Clin Med. 1999;134(3):194-214.
 Lutay MI. On the clinical classification dislipoproteinemia. Ukrainian journal of cardiology. 2003. 3(4):9-16.
 Klimov AN, Nikul'cheva NG. Lipoproteins, dyslipidemia and atherosclerosis. Leningrad: Meditsina, 1984. 166 p.
 Tangirala RK, Tsukamoto K, Chun SH, Usher D, Pure E, Rader DJ. Regression of atherosclerosis induced by liver-directed gene transfer of apolipoprotein A-I in mice. Circulation. 1999;100(17):1816-22.
 Muzyczka N. Use of adeno-associated virus as a general transduction vector for mammalian cells. Curr Top Microbiol Immunol. 1992;158:97-129.
 Conway JE, Zolotukhin S, Muzyczka N, Hayward GS, Byrne BJ. Recombinant adeno-associated virus type 2 replication and packaging is entirely supported by a herpes simplex virus type 1 amplicon expressing Rep and Cap. J Virol. 1997;71(11):8780-9.
 Rayfield M, Michaels GS, Feldmann R, Muzyczka N. Comparison of the DNA sequence and secondary structure of the herpes simplex virus L/S junction and the adeno-associated virus terminal repeat. J Theor Biol. 1985;115(4):477-94.
 Kotin RM, Siniscalco M, Samulski RJ, Zhu XD, Hunter L, Laughlin CA, McLaughlin S, Muzyczka N, Rocchi M, Berns KI. Site-specific integration by adeno-associated virus. Proc Natl Acad Sci U S A. 1990;87(6):2211-5.
 Yang CC, Xiao X, Zhu X, Ansardi DC, Epstein ND, Frey MR, Matera AG, Samulski RJ. Cellular recombination pathways and viral terminal repeat hairpin structures are sufficient for adeno-associated virus integration in vivo and in vitro. J Virol. 1997;71(12):9231-47.
 Rizzuto G, Gorgoni B, Cappelletti M, Lazzaro D, Gloaguen I, Poli V, Sgura A, Cimini D, Ciliberto G, Cortese R, Fattori E, La Monica N. Development of animal models for adeno-associated virus site-specific integration. J Virol. 1999;73(3):2517-26.
 Lalwani AK, Han JJ, Walsh BJ, Zolotukhin S, Muzyczka N, Mhatre AN. Green fluorescent protein as a reporter for gene transfer studies in the cochlea. Hear Res. 1997;114(1-2):139-47.
 Peel AL, Zolotukhin S, Schrimsher GW, Muzyczka N, Reier PJ. Efficient transduction of green fluorescent protein in spinal cord neurons using adeno-associated virus vectors containing cell type-specific promoters. Gene Ther. 1997;4(1):16-24.
 Toporova EK, Irodov DM, Sukhorada YeM, Ruban TA, Kozel YuN, Andriyenko VI, Novikova SN, Kordyum VA. Using genes of fluorescent proteins for visualization of gene expression in living cells in vitro and in vivo. X Jubilee Int. conf.: New information technologies in medicine and ecology. Yalta; Gursuf, 2002; 301-3.
 Maniatis T., Fritsch E. F., Sambrook J. Molecular cloning: a laboratory manual New York: Cold Spring Harbor Lab. publ., 1982 545 p.
 Botlag D.M., Rozycki D.M., Edelstein S.J. Protein Methods, New York 1996 p. 415.
 Shulzhenko V. N., Lukash L. L., Shulyak L. N., Usenko E. V., Kordyum V. A. Cloning of human ApoA1 gene and its expression in murine fibroblasts. Biopolym. Cell. 1989; 5(5):105-107
 Thomas M, Klibanov AM. Non-viral gene therapy: polycation-mediated DNA delivery. Appl Microbiol Biotechnol. 2003;62(1):27-34.
 Erbacher P, Bettinger T, Belguise-Valladier P, Zou S, Coll JL, Behr JP, Remy JS. Transfection and physical properties of various saccharide, poly(ethylene glycol), and antibody-derivatized polyethylenimines (PEI). J Gene Med. 1999;1(3):210-22.
 Shpilevaya S. P., Andrienko V. I., Toporova H. K., Eliseeva G. S., Ajamian F., Zharova L. G., Likhacheva L. I., Kordium V. A. The expression of pEGFP plasmid genes in mouse hepatocyles in vivo. Biopolym. Cell. 1999; 15(3):241-246
 Kozel Y., Suhorada H., Ruban T., Shpylevaya S., Titok T., Andrienko V., Irodov D., (...), Toporova H. Monitoring of Gene Expression With the Use of Marker GFP and DsRed Genes In Mammalian Cells Abstr. book Conf. for Students, PhD students and Young Scientist on Mol. Biol, and Genet. Kyiv, 2001
 Kozel Y., Suhorada H., Ruban T., Shpylevaya S., Titok T., Andrienko V., Irodov D., Toporova H. Visualization of gene delivery systems. Abstracts III Ukr. conf. Young scientists dedicated to the memory of academician. V. Frolkis. Kyiv, 2002:93.
 Kathy D. Antibodies. Moscow, Mir, 1991. 287.
 Frolkis V. V., Kordyum V. A., Novikova S. N., Bogatskaya L. N., Irodov D. M., Likhacheva L. I., Potapenko R. I., Mozguhina T. G., Bitner M. K. The influence of the implanted cloning human's ApoAI gene on the development of rabbit' hypercholesterinemy. Biopolym. Cell. 1994; 10(6):98-105