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Adenovirus-mediated apo(a)-antisense-RNA expression efficiently inhibits apo(a) synthesis in vitro and in vivo

Gene Therapy volume 8, pages 425–430 (2001)Cite this article

Abstract

Apo(a) is a very atherogenic plasma protein without apparent function, which is highly expressed in humans. The variation in plasma Lp(a) concentration among individuals is considerable. Approximately 10–15% of the white population exhibit plasma Lp(a) concentrations above the atherogenic cut-off value of approximately 30 mg/dl. Since there is currently no safe way of treating those patients with drugs, we have tested the possibility of interfering with apo(a) biosynthesis by adenovirus-mediated expression of antisense apo(a) mRNA comprising the 5′ UTR, the signal sequence and the first three kringles of native apo(a). Transduction of rat hepatoma McA RH 7777 cells which stably expressed apo(a) with 18 kringle IV (KIV) domains with apo(a)-antisense adenovirus (AS-Ad) at multiplicity of infection (MOI) of 30 reduced apo(a) synthesis to 23% as compared with control cells. As apo(a) is not synthesized in laboratory animals, we induced biosynthesis of the N-terminal fragments of apo(a) in mice by adenovirus-mediated gene transfer. Cotransduction of these mice with AS-Ad, which expressed up to eight times higher amounts of apo(a) than stable transgenic apo(a) mice, led to an almost complete disappearance of apo(a) from plasma. We conclude that the proposed AS-construct is very efficient in interfering with apo(a) biosynthesis in vivo. The strategy of inducing the synthesis of a nonexpressed protein followed by knocking it out by AS technology may also be applicable to other systems.

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References

  1. Kostner GM, Krempler F . Lipoprotein(a) Curr Opin Lipidol 1992 3: 279–284

    Article  CAS  Google Scholar 

  2. Kostner GM et al. Lipoprotein Lp(a) and the risk for myocardial infarction Atherosclerosis 1981 38: 51–61

    Article  CAS  PubMed  Google Scholar 

  3. Budde T et al. Plasma Lp(a) levels correlate with number, severity, and length-extension of coronary lesions in male patients undergoing coronary arteriography for clinically suspected coronary atherosclerosis Arterioscler Thromb 1994 14: 1730–1736

    Article  CAS  PubMed  Google Scholar 

  4. Rader DJ et al. The inverse association of plasma lipoprotein(a) concentrations with apolipoprotein(a) isoform size is not due to differences in Lp(a) catabolism but to differences in production rate J Clin Invest 1994 93: 2758–2763

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Utermann G . Lipoprotein(a). In: Scriver CR, Beaudet AL, Sly WS, Valle D (eds) The Metabolic and Molecular Bases of Inherited Disease McGraw Hill: New York 1995 1887–1912

    Google Scholar 

  6. McLean JW et al. cDNA sequence of human apo-lipoprotein(a) is homologous to plasminogen Nature 1987 330: 132–137

    Article  CAS  PubMed  Google Scholar 

  7. Morrisett J, Gaubatz J, Knapp R, Guevara JG . Structural properties of apo(a): a major apoprotein of human lipoprotein(a). In: Scanu A (ed.) Lipoprotein(a) Academic Press: San Diego 1990 53–72

    Google Scholar 

  8. Lackner C, Cohen JC, Hobbs HH . Molecular definition of the extreme size polymorphism in apolipoprotein(a) Hum Mol Genet 1993 2: 933–940

    Article  CAS  PubMed  Google Scholar 

  9. Van der Hoeck YY et al. The apoprotein(a) kringle IV repeats which differ from the major repeat kringle are present in variably-sized isoforms Hum Mol Genet 1993 2: 361–366

    Article  Google Scholar 

  10. Craig WY et al. Lipoprotein(a) as a risk factor for ischemic heart disease: metaanalysis of prospective studies Clin Chem 1998 44: 2301–2306

    CAS  PubMed  Google Scholar 

  11. Lawn R et al. Atherogenesis in transgenic mice expressing human apolipoprotein(a) Nature 1992 360: 670–672

    Article  CAS  PubMed  Google Scholar 

  12. Callow M et al. Atherogenesis in transgenic mice with human apolipoprotein B and lipoprotein(a) J Clin Invest 1995 96: 1639–1646

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Grainger DJ et al. Activation of transforming growth factor-β is inhibited in transgenic apolipoprotein(a) mice Nature 1994 370: 460–462

    Article  CAS  PubMed  Google Scholar 

  14. Lawn RM et al. Feedback mechanism of focal vascular lesion formation in transgenic apolipoprotein(a) mice J Biol Chem 1996 271: 31367–31371

    Article  CAS  PubMed  Google Scholar 

  15. Kostner GM et al. HMG CoA reductase inhibitors lower LDL cholesterol without reducing Lp(a) levels Circulation 1989 80: 1313–1319

    Article  CAS  PubMed  Google Scholar 

  16. Carlson LA, Hamsten A, Asplund A . Pronounced lowering of serum levels of lipoprotein Lp(a) in hyperlipidaemic subjects treated with nicotinic acid J Intern Med 1989 226: 271–276

    Article  CAS  PubMed  Google Scholar 

  17. Seed M et al. The effect of nicotinic acid and acipimox on lipoprotein(a) concentration and turnover Atherosclerosis 1993 101: 61–68

    Article  CAS  PubMed  Google Scholar 

  18. Albers JJ et al. Reduction of lecithin cholesterol acyltransferase, apolipoprotein D and the LP(a) lipoprotein with the anabolic sterol stanazolol Biochim Biophys Acta 1994 795: 293–296

    Article  Google Scholar 

  19. Kagawa A et al. Aspirin reduces apolipoprotein(a) (apo((a)) production in human hepatocytes by suppression of apo(a) gene transcription J Biol Chem 1999 274: 34111–34115

    Article  CAS  PubMed  Google Scholar 

  20. Robertson HD . Escherichia coli ribonuclease III cleavage sites Cell 1982 30: 669–672

    Article  CAS  PubMed  Google Scholar 

  21. Abou Elela S, Igel H, Ares JrM . RNase III cleaves eukaryotic preribosomal RNA at a U3 snoRNP-dependent site Cell 1996 85: 115–124

    Article  Google Scholar 

  22. Rotondo G, Frendewey D . Purification and characterization of the Pac1 ribonuclease of Schizosaccharomyces pombe Nucleic Acids Res 1996 24: 2377–2386

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Lozier JN, Metzger ME, Donahue RE, Morgan RA . Adenovirus-mediated expression of human coagulation factor IX in the rhesus macaque is associated with dose-limiting toxicity Blood 1999 94: 3968–3975

    CAS  PubMed  Google Scholar 

  24. Lehrman S . Virus treatment questioned after gene therapy death Nature 1999 401: 517–518

    Article  CAS  PubMed  Google Scholar 

  25. Kochanek S . High-capacity adenoviral vectors for gene transfer and somatic gene therapy Hum Gene Ther 1999 10: 2451–2459

    Article  CAS  PubMed  Google Scholar 

  26. Morral N et al. High doses of a helper-dependent adenoviral vector yield supraphysiological levels of α1-antitrypsin with negligible toxicity Hum Gene Ther 1998 9: 2709–2716

    Article  CAS  PubMed  Google Scholar 

  27. Sugano M, Makino N . Changes in plasma lipoprotein cholesterol levels by antisense oligodeoxynucleotides against cholesteryl ester transfer protein in cholesterol-fed rabbits J Biol Chem 1995 271: 19080–19083

    Article  Google Scholar 

  28. Frank S, Kostner GM . The role of apo(a) kringle IVs in the assembly of lipoprotein(a) Protein Eng 1997 3: 291–298

    Article  Google Scholar 

  29. Frank S et al. Effect of tranexamic acid and ∂-aminovaleric acid on lipoprotein(a) metabolism in transgenic mice Biochim Biophys Acta 1999 1438: 99–110

    Article  CAS  PubMed  Google Scholar 

  30. Kobayashi K et al. Reversal of hypercholesterolemia in low density lipoprotein receptor knockout mice by adenovirus-mediated gene transfer of the very low density lipoprotein receptor J Biol Chem 1996 271: 6852–6860

    Article  CAS  PubMed  Google Scholar 

  31. McGrory WJ, Bautista DS, Graham FL . A simple technique for the rescue of early region I mutations into infectious human adenovirus type 5 Virology 1988 163: 614–617

    Article  CAS  PubMed  Google Scholar 

  32. Graham FL, van der Eb AJ . A new technique for the assay of infectivity of human adenovirus IV DNA Virology 1973 52: 456–467

    Article  CAS  PubMed  Google Scholar 

  33. Teng B et al. Adenovirus-mediated gene transfer of a rat apolipoprotein B mRNA-editing protein in mice virtually eliminates apolipoprotein B-100 and normal low density lipoprotein production J Biol Chem 1994 269: 29395–29404

    CAS  PubMed  Google Scholar 

  34. Frank S, Durovic S, Kostner GM . Structural requirements of apo(a) for the lipoprotein(a) assembly Biochem J 1994 304: 27–30

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was supported by grants from the Austrian Research Foundation SFB 702 and the Austrian National Bank ÖNB #7475. The technical assistance of Harald Grillhofer, Margit Eichholzer and Anton Ibovnik is highly appreciated.

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Authors and Affiliations

  1. Institute of Medical Biochemistry and Medical Molecular Biology, Karl-Franzens-University Graz, Harrachgasse 21, 3rd Floor, Graz, 8010, Austria

    S Frank, M Gauster, J Strauss, A Hrzenjak & G M Kostner

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  1. S Frank
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  2. M Gauster
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  4. A Hrzenjak
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  5. G M Kostner
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Frank, S., Gauster, M., Strauss, J. et al. Adenovirus-mediated apo(a)-antisense-RNA expression efficiently inhibits apo(a) synthesis in vitro and in vivo. Gene Ther 8, 425–430 (2001). https://doi.org/10.1038/sj.gt.3301434

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