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2001 | 48 | 4 | 1061-1076
Article title

Antisense hairpin loop oligonucleotides as inhibitors of expression of multidrug resistance-associated protein 1: Their stability in fetal calf serum and human plasma.

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EN
Abstracts
EN
Multidrug resistance-associated protein (MRP1) is a transmembrane pump protein responsible for the efflux of chemotherapeutic drugs, an important cause of anticancer treatment failure. Trying to circumvent MRP-mediated resistance we designed and synthesized hairpin loops forming antisense oligodeoxyribonucleotides (ODNs), both phosphodiesters (PO-ODNs) and their phosphorothioate analogues (PS-ODNs), to reduce the protein expression by targeting its mRNA in a sequence specific manner. Melting temperature measurements as well as polyacrylamide gel electrophoresis supported the preferential formation of a secondary structure, which was expected to protect ODNs against 3'-exonuclease degradation. ODNs and PS-ODNs designed in this work were successfully tested as antisense inhibitors of the expression of MRP1 in the leukaemia HL60/ADR cell line. Foreseeing the necessity to perform clinical studies with such ODNs we investigated their stability against the 3'-exonuclease activity of fetal calf serum and human plasma. Under the conditions, corresponding to physiological ones, we observed high stability of hairpin loop forming ODNs, especially those containing longer (e.g. 7 base pair) stems. Comparative studies on the stability of chemically unmodified hairpin loop forming ODNs and their PS-counterparts indicated that endonuclease activity did not play any important role in the process of their nucleolytic degradation. Our studies provide strong evidence for high stability of chemically unmodified hairpin loop ODNs, making them an attractive alternative to phosphorothioate analogues commonly used in antisense strategy.
Year
Volume
48
Issue
4
Pages
1061-1076
Physical description
Dates
published
2001
received
2001-07-4
revised
2001-10-16
accepted
2001-11-3
References
  • 1. Zamecnik, P.C. & Stephenson, M.L. (1978) Inhibition of Rous sarcoma virus replication and cell transformation by a specific oligodeoxynucleotide. Proc. Natl. Acad. Sci. U.S.A. 75, 285-288.
  • 2. Agrawal, S. & Zhao, Q. (1998) Antisense therapeutics. Curr. Opin. Chem. Biol. 2, 519-528.
  • 3. Ma, L. & Calvo, F. (1996) Recent status of the antisense oligonucleotide approach in oncology. Fundam. Clin. Pharmacol. 10, 97-115.
  • 4. Calabretta, B. & Skórski, T. (1997) Targeting c-myc in leukemia. Anticancer Drug Des. 12, 373-381.
  • 5. Gewirtz, A.M. (1998) Antisense oligonucleotide therapeutics for human leukemia. Curr. Opin. Hematol. 5, 59-71.
  • 6. Crook, S.T. (1998) Basic principles of antisense therapeutics; in Antisense Research and Application (Crooke, S.T., ed.) pp. 1-50, Springer-Verlag, Berlin, Heidelberg.
  • 7. Eder, P.S., DeVine, R.J., Dagle, J.M. & Walder, J.A. (1991) Substrate specificity and kinetics of degradation of antisense oligonucleotides by a 3’-exonuclease in plasma. Antisense Res. Dev. 1, 141-151.
  • 8. De Clercq, E., Eckstein, E. & Merigan, T.C. (1969) Interferon induction increased through chemical modification of synthetic polyribonucleotide. Science 165, 1137-1139.
  • 9. Cohen, J.S. (1993) Phosphorothioate oligodeoxynucleotides; in Antisense Research and Application (Crooke, S.T., Lebleu, B., eds.) pp.205-222, CRC Press, Boca Raton.
  • 10. Miller, P.S., McParland, K.B., Jayaraman, K. & Ts’o, P.O. (1981) Biochemical and biological effects of nonionic acid methylphosphonate. Biochemistry 20, 1874-1880.
  • 11. Cook, P.D. (1993) Medicinal chemistry strategies for antisense research; in Antisense Research and Application (Crooke, S.T., Lebleu, B., eds.) pp. 51-102, CRC Press, Boca Raton.
  • 12. Nilsen, P.E., Engholm, M., Berg, R.H. & Buchardt, O. (1991) Sequence-selective recognition of DNA by strand displacement with a thymine-substituted polyamide. Science 254, 1497-1500.
  • 13. Gao, H., Yang, M., Patel, R. & Cook, A.F. (1995) Circularization of oligonucleotides by disulfide bridge formation. Nucleic Acids Res. 23, 2025-2029.
  • 14. Azhayeva, E., Azhayev, A., Auriola, S., Tengvall, U., Urtti, A. & Lonnberg, H. (1997) Inhibitory properties of double helix forming circular oligonucleotides. Nucleic Acids Res. 25, 4954-4961.
  • 15. Lim, C.L. & Hunt, A. (1997) Synthesis of DNA dumbbells: Chemical vc. enzymatic ligation of self-complementary oligonucleotides. Nucleosides Nucleotides 16, 41-51.
  • 16. Yamakawa, H., Hosono, K., Ishibashi, T., Takai, K. & Takaku, H. (1995) Properties of nicked and circular dumbbell RNA-DNA chimeric oligonucleotides. Nucleic Acids Symp. Ser. 34, 133-134.
  • 17. Prakash, G. & Kool, E.T. (1991) Molecular recognition by circular oligonucleotides. Strong binding of single-stranded DNA and RNA. J. Chem. Soc., Chem. Commun. 1161-1163.
  • 18. Ashley, G. & Kushlan, D.M. (1991) Chemical synthesis of oligodeoxynucleotide dumbbells. Biochemistry 30, 2927-2933.
  • 19. Tang, J.-Y., Temsamani, J. & Agrawal, S. (1993) Self-stabilized antisense oligonucleotide phosphorothioates: Properties and anti-HIV activity. Nucleic Acids Res. 21, 2729-2735.
  • 20. Kool, E.T. (1998) Recognition of DNA, RNA, and proteins by circular oligonucleotides. Acc. Chem. Res. 31, 502-510.
  • 21. Lautier, D., Canitrot, Y., Deeley, R.G. & Cole, S.P.C. (1996) Multidrug resistance mediated by the multidrug resistance protein (MRP) gene. Biochem. Pharmacol. 52, 967-977.
  • 22. Nooter, K., Burger, H. & Stoten, G. (1996) Multidrug resistance-associated protein (MRP) in haematological malignancies. Leuk. Lymphoma 20, 381-387.
  • 23. Almquist, K.C., Loe, D.W., Hipfner, D.R., Mackie, J.E., Cole, S.P.C. & Deeley, R.G. (1995) Characterisation of the 190 kDa multidrug resistance protein (MRP) in drug selected and transfected human tumour cells. Cancer Res. 55, 102-110.
  • 24. Gendaszewska, E., Rębowski, G., Soszyński, M., Bartosz, G. & Niewiarowski, W. (1999) Inhibition of multidrug resistance-associated protein (MRP) expression in HL60/ADR cell line by means of phosphorothioate oligonucleotides (British Cancer Research Meeting. Edinburgh, United Kingdom, 11-14 July 1999) Brit. J. Cancer 80, Suppl 2, 1-113.
  • 25. Niewiarowski, W., Gendaszewska, E, Rębowski,G., Wójcik, M., Mikołajczyk, B, Goss, W., Soszyński, M. & Bartosz, G. (2000) Multidrug resistance-associated protein - reduction of expression in human leukaemia cells by antisense phosphorothioate oligonucleotides. Acta Biochim. Polon. 47, 1183- 1188.
  • 26. Beaucage, S.L. & Iyer, R.P. (1993) The synthesis of modified oligonucleotides by the phosphoramidite approach and their applications. Tetrahedron 49, 6123-6194.
  • 27. Stec, W.J., Zon, G., Egan, W. & Stec, B. (1984) Automated solid-phase synthesis, separation, and stereochemistry of phosphorothioate analogues of oligodeoxyribonucleotides. J. Am. Chem. Soc. 106, 6077-6080.
  • 28. Stec, W.J., Uznański, B., Wilk, A., Hirschbein, B.L., Fearon, K.L. & Bergot, B.J. (1993) Bis( O,O-diisopropoxy phosphinothioyl)disulfide-A highly efficient sulfurizing reagent for cost-effective synthesis of oligo(nucleoside phosphorothioate)s. Tetrahedron Lett. 34, 5317-5320.
  • 29. Brown T. & Brown D.J.S. (1991) in Oligonucleotides and Analogues. A Practical Approach (Eckstein F., ed.) pp. 20-86, Oxford University Press, U.K.
  • 30. Lowry, M.O.H., Rosebrough, N.J., Farr, R.L. & Randal, R.J. (1951)Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265-275.
  • 31. Laemmli, U.A. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-684.
  • 32. Hansen, M.B., Nielsen, S.E., Berg, K. (1989) Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J. Immunol. Methods 119, 203-210.
  • 33. Zucker, M. (1989) On finding all suboptimal foldings of an RNA molecule. Science 244, 48-52.
  • 34. Jaeger, J.A., Turner, D.H., Zucker, M. (1989) Improved predictions of secondary structures for RNA. Proc. Natl. Acad. Sci. U.S.A. 86, 7706-7710.
  • 35. Mathews, D.H., Sabina, J., Zuker, M. & Turner, D.H. (1999) Expanded sequence dependence of thermodynamic parameters improves prediction of RNA secondary structure. J. Mol. Biol. 288, 911-940.
  • 36. Davis, T.M., McFail-Isom, L., Keane, E. & Williams, L.D. (1998) Melting of DNA hairpin without hyperchromism. Biochemistry 37, 6975-6978.
  • 37. Stewart, A.J., Canitrot, Y., Baracchini, E., Dean, N.M., Deeley, R.G. & Cole, S.P. (1996) Reduction of expression of the multidrug resistance protein (MRP) in human tumor cells by antisense phosphorothioate oligonucleotides. Biochem. Pharmacol. 51(4), 461-469.
  • 38. Stec, W.J., Cierniewski, C.S., Okruszek, A., Kobylańska, A., Pawłowska, Z., Koziołkiewicz, M., Pluskota, E., Maciaszek, A., Rębowska, B. & Stasiak, M. (1997) Stereodependent inhibition of plasm inogen activator inhibitor type 1 by phosphorothioate oligonucleotides; proof of sequence specificity in cell culture and in vivo rats experiments . Antisense Nucleic Acid Drug Dev. 7, 567-573.
  • 39. Stein, C.A. (1996) Phosphorothioate antisense oligodeoxynucleotides: Questions of specificity. Trends Biotechnol. 14, 147-149.
  • 40. Levin, A.A., Monteith, D.K., Leeds, J.M., Nicklin, P.L., Geary, R.S., Butler, M., Templin, M.V. & Henry, S.P. (1998) Toxicity of oligodeoxynucleotide therapeutic agents; in: Antisense Research and Application; Handbook of Experimental Pharmacology. Springer- Verlag. Berlin, Heidelberg, 131, 169-215.
  • 41. Krieg, A.M. (1996) Immune stimulation by oligonucleotides; in Antisense Research and Application (Crooke, S.T., ed.), Handbook of Experimental Pharmacology, Berlin, Heidelberg, 131, 243-262.
  • 42. Azhayeva, E., Azhayev, A., Guzaev, A. & Lonnberg, H. (1995) Selective binding of looped oligonucleotides to a single-stranded DNA and its influence on replication in vitro. Nucleic Acids Res. 23, 4255-4261.
  • 43. Maksimenko, A.V., Gottikh, M.B., Helin, V., Shabarova, Z.A. & Malvy, C. (1999) Physicochemical and biological properties of antisense phosphodiester oligonucleotides with various secondary structures. Nucleosides Nucleotides 18, 2071-2091
  • 44. Stewart, A.J., Canitrot, Y., Baracchini, E., Dean, N.M, Deeley, R.G. & Cole, S.P.C. (1996) Reduction of expression of the multidrug resistance protein (MRP) in human tumor cells by antisense phosphorothioate oligonucleotides. Biochem. Pharmacol. 51, 461-469.
  • 45. Dassow, H., Lassner, D., Remke, H. & Preoss, R. (2000) Modulation of multidrug resistance in human leukaemia cells with MDR1-targeted antisense oligonucleotides using variable treatment schedules. Int. J. Clin. Pharmacol. Ther. 38, 209-216.
  • 46. Cole, S.P.C., Sparks, K., Fraser, K., Loe, D., Guart, C., Wilson, G. & Deeley, R.G. (1994) Pharmacological characterization of multidrug resistant MRP-transfected human tumor cells. Cancer Res. 54, 5902-5910.
  • 47. Park, W.S., Miyano-Kurosaki, N., Abe, T., Takai, K., Yamamoto, N. & Takaku, H. (2000) Inhibition of HIV-1 replication by a new type of circular dumbbell RNA/DNA chimeric oligonucleotides. Biochem. Biophys. Res. Commun. 270, 953-960.
Document Type
Publication order reference
YADDA identifier
bwmeta1.element.bwnjournal-article-abpv48i4p1061kz
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