Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1996 May 14;93(10):4903–4906. doi: 10.1073/pnas.93.10.4903

In vivo suppression of the renal Na+/Pi cotransporter by antisense oligonucleotides.

R Oberbauer 1, G F Schreiner 1, J Biber 1, H Murer 1, T W Meyer 1
PMCID: PMC39377  PMID: 8643501

Abstract

A 20-mer phosphorothioate oligonucleotide (AS1) was designed to hybridize to the message for the rat kidney sodium phosphate cotransporter NaPi-2 close to the translation initiation site. Single intravenous doses of this oligonucleotide were given to rats maintained on a low phosphorus diet to increase NaPi-2 expression. At 3 days after oligonucleotide infusion, rats receiving 2.5 micromol of AS1 exhibited a reduction in renal NaPi-2 to cyclophilin mRNA ratio by 40% +/- 17%, and rats receiving 7.5 micromol of AS1 exhibited a reduction in NaPi-2 to cyclophilin mRNA ratio by 46% +/- 21%. Reversed-sequence AS1 was without effect. The higher dose of 7.5 micromol of AS1 also reduced the rate of phosphate uptake into renal brush border membrane vesicles and the expression of NaPi-2 protein detected by Western blotting in these vesicles. Reversed sequence AS1 was again without effect on these parameters. These results suggest that systemically infused oligonucleotides can exert antisense effects in the renal proximal tubule.

Full text

PDF
4903

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Agrawal S., Temsamani J., Tang J. Y. Pharmacokinetics, biodistribution, and stability of oligodeoxynucleotide phosphorothioates in mice. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7595–7599. doi: 10.1073/pnas.88.17.7595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Biber J., Stieger B., Haase W., Murer H. A high yield preparation for rat kidney brush border membranes. Different behaviour of lysosomal markers. Biochim Biophys Acta. 1981 Oct 2;647(2):169–176. doi: 10.1016/0005-2736(81)90243-1. [DOI] [PubMed] [Google Scholar]
  3. Brown C. D., Bodmer M., Biber J., Murer H. Sodium-dependent phosphate transport by apical membrane vesicles from a cultured renal epithelial cell line (LLC-PK1). Biochim Biophys Acta. 1984 Jan 25;769(2):471–478. doi: 10.1016/0005-2736(84)90332-8. [DOI] [PubMed] [Google Scholar]
  4. Christensen E. I., Nielsen S. Structural and functional features of protein handling in the kidney proximal tubule. Semin Nephrol. 1991 Jul;11(4):414–439. [PubMed] [Google Scholar]
  5. Cohen J. S. Gene-mimetic substances: drugs designed to intervene in gene expression. Adv Pharmacol. 1994;25:319–339. doi: 10.1016/s1054-3589(08)60436-6. [DOI] [PubMed] [Google Scholar]
  6. Cossum P. A., Sasmor H., Dellinger D., Truong L., Cummins L., Owens S. R., Markham P. M., Shea J. P., Crooke S. Disposition of the 14C-labeled phosphorothioate oligonucleotide ISIS 2105 after intravenous administration to rats. J Pharmacol Exp Ther. 1993 Dec;267(3):1181–1190. [PubMed] [Google Scholar]
  7. Custer M., Lötscher M., Biber J., Murer H., Kaissling B. Expression of Na-P(i) cotransport in rat kidney: localization by RT-PCR and immunohistochemistry. Am J Physiol. 1994 May;266(5 Pt 2):F767–F774. doi: 10.1152/ajprenal.1994.266.5.F767. [DOI] [PubMed] [Google Scholar]
  8. Danielson P. E., Forss-Petter S., Brow M. A., Calavetta L., Douglass J., Milner R. J., Sutcliffe J. G. p1B15: a cDNA clone of the rat mRNA encoding cyclophilin. DNA. 1988 May;7(4):261–267. doi: 10.1089/dna.1988.7.261. [DOI] [PubMed] [Google Scholar]
  9. Dean N. M., McKay R. Inhibition of protein kinase C-alpha expression in mice after systemic administration of phosphorothioate antisense oligodeoxynucleotides. Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11762–11766. doi: 10.1073/pnas.91.24.11762. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gao W. Y., Storm C., Egan W., Cheng Y. C. Cellular pharmacology of phosphorothioate homooligodeoxynucleotides in human cells. Mol Pharmacol. 1993 Jan;43(1):45–50. [PubMed] [Google Scholar]
  11. Kempson S. A., Lötscher M., Kaissling B., Biber J., Murer H., Levi M. Parathyroid hormone action on phosphate transporter mRNA and protein in rat renal proximal tubules. Am J Physiol. 1995 Apr;268(4 Pt 2):F784–F791. doi: 10.1152/ajprenal.1995.268.4.F784. [DOI] [PubMed] [Google Scholar]
  12. Ku N. O., Michie S., Oshima R. G., Omary M. B. Chronic hepatitis, hepatocyte fragility, and increased soluble phosphoglycokeratins in transgenic mice expressing a keratin 18 conserved arginine mutant. J Cell Biol. 1995 Dec;131(5):1303–1314. doi: 10.1083/jcb.131.5.1303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Leonetti J. P., Mechti N., Degols G., Gagnor C., Lebleu B. Intracellular distribution of microinjected antisense oligonucleotides. Proc Natl Acad Sci U S A. 1991 Apr 1;88(7):2702–2706. doi: 10.1073/pnas.88.7.2702. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Magagnin S., Werner A., Markovich D., Sorribas V., Stange G., Biber J., Murer H. Expression cloning of human and rat renal cortex Na/Pi cotransport. Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):5979–5983. doi: 10.1073/pnas.90.13.5979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Morishita R., Gibbons G. H., Ellison K. E., Nakajima M., Zhang L., Kaneda Y., Ogihara T., Dzau V. J. Single intraluminal delivery of antisense cdc2 kinase and proliferating-cell nuclear antigen oligonucleotides results in chronic inhibition of neointimal hyperplasia. Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8474–8478. doi: 10.1073/pnas.90.18.8474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Oberbauer R., Schreiner G. F., Meyer T. W. Renal uptake of an 18-mer phosphorothioate oligonucleotide. Kidney Int. 1995 Oct;48(4):1226–1232. doi: 10.1038/ki.1995.406. [DOI] [PubMed] [Google Scholar]
  18. Rappaport J., Hanss B., Kopp J. B., Copeland T. D., Bruggeman L. A., Coffman T. M., Klotman P. E. Transport of phosphorothioate oligonucleotides in kidney: implications for molecular therapy. Kidney Int. 1995 May;47(5):1462–1469. doi: 10.1038/ki.1995.205. [DOI] [PubMed] [Google Scholar]
  19. Ratajczak M. Z., Kant J. A., Luger S. M., Hijiya N., Zhang J., Zon G., Gewirtz A. M. In vivo treatment of human leukemia in a scid mouse model with c-myb antisense oligodeoxynucleotides. Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):11823–11827. doi: 10.1073/pnas.89.24.11823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Simons M., Edelman E. R., DeKeyser J. L., Langer R., Rosenberg R. D. Antisense c-myb oligonucleotides inhibit intimal arterial smooth muscle cell accumulation in vivo. Nature. 1992 Sep 3;359(6390):67–70. doi: 10.1038/359067a0. [DOI] [PubMed] [Google Scholar]
  21. Stein C. A., Cheng Y. C. Antisense oligonucleotides as therapeutic agents--is the bullet really magical? Science. 1993 Aug 20;261(5124):1004–1012. doi: 10.1126/science.8351515. [DOI] [PubMed] [Google Scholar]
  22. Stoll R., Kinne R., Murer H. Effect of dietary phosphate intake on phosphate transport by isolated rat renal brush-border vesicles. Biochem J. 1979 Jun 15;180(3):465–470. doi: 10.1042/bj1800465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tenenhouse H. S., Werner A., Biber J., Ma S., Martel J., Roy S., Murer H. Renal Na(+)-phosphate cotransport in murine X-linked hypophosphatemic rickets. Molecular characterization. J Clin Invest. 1994 Feb;93(2):671–676. doi: 10.1172/JCI117019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wagner R. W. Gene inhibition using antisense oligodeoxynucleotides. Nature. 1994 Nov 24;372(6504):333–335. doi: 10.1038/372333a0. [DOI] [PubMed] [Google Scholar]
  26. Werner A., Kempson S. A., Biber J., Murer H. Increase of Na/Pi-cotransport encoding mRNA in response to low Pi diet in rat kidney cortex. J Biol Chem. 1994 Mar 4;269(9):6637–6639. [PubMed] [Google Scholar]
  27. Yakubov L. A., Deeva E. A., Zarytova V. F., Ivanova E. M., Ryte A. S., Yurchenko L. V., Vlassov V. V. Mechanism of oligonucleotide uptake by cells: involvement of specific receptors? Proc Natl Acad Sci U S A. 1989 Sep;86(17):6454–6458. doi: 10.1073/pnas.86.17.6454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Zamecnik P., Aghajanian J., Zamecnik M., Goodchild J., Witman G. Electron micrographic studies of transport of oligodeoxynucleotides across eukaryotic cell membranes. Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3156–3160. doi: 10.1073/pnas.91.8.3156. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES