K+ channel antisense oligodeoxynucleotides inhibit cytokine-induced expansion of human hemopoietic progenitors (original) (raw)

Abstract

Primitive human hemopoietic progenitor cells identified by surface membrane markers CD33−CD34+ are capable of expansion into lineage-restricted precursors following in vitro stimulation by hemopoietic regulators such as stem cell factor (SCF) and interleukin-3 (IL-3). In search of ionic currents involved in cytokine-induced progenitor cell growth and differentiation, human umbilical cord blood CD33−CD34+ cells were subjected to perforated patch-clamp recordings following overnight incubation with SCF and/or IL-3. An inward rectifying potassium channel (Kir) was found in 33% of control unstimulated cells, in 34% of cells incubated with IL-3, in 31 % of cells incubated with SCF and in 75% of cells incubated with IL-3 plus SCE Kir activity increased with elevation of extracellular potassium and was blocked by extracellular Cs+ or Ba2+. Antisense oligodeoxynucleotides directed against Kir blocked both mRNA and functional expression of Kir channels. Kir antisense also inhibited the in vitro expansion of cytokine-stimulated CD33−CD34+ cells into erythroid (BFU-E) and myeloid (GM-CFU) progenitors in 7-day suspension cultures. Extracellular Cs+ or Ba2+ induced a similar degree of inhibition (40–60%) of progenitor cell generation. These findings strongly suggest an essential role for Kir in the process of cytokine-induced primitive progenitor cell growth and differentiation.

Access this article

Log in via an institution

Subscribe and save

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Discover the latest articles and news from researchers in related subjects, suggested using machine learning.

References

  1. Andrews RG, Singer JW, Bernstein ID (1989) Precursors of colony-forming cells in humans can be distinguished from colony-forming cells by expression of the CD33 and CD34 antigens and light scatter properties. J Exp Med 169:1721–1731
    Google Scholar
  2. Bernstein ID, Andrews RG, Zsebo KM (1991) Recombinant human stem cell factor enhances the formation of colonies by CD34+ and CD34+lin- cells, and the generation of colonyforming cell progeny from CD34+lin- cells cultured with interleukin-3, granulocyte colony-stimulating factor, or granulocyte-macrophage colony-stimulating factor. Blood 77: 2316–2321
    Google Scholar
  3. Brizzi MF, Zini MG, Aronica MG, Blechman JM, Yarden Y, Pegoraro L (1994) Convergence of signaling by interleukin-3, granulocyte-macrophage colony-stimulating factor, and mast cell growth factor on JAK2 tyrosine kinase. J Biol Chem 269:31680–31684
    Google Scholar
  4. Chandy KG, DeCoursey TE, Cahalan MD, Gupta S (1985) Ion channels in lymphocytes (Review). J Clin Immunol 5:1–6
    Google Scholar
  5. Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159
    Google Scholar
  6. Decoursey TE, Chandy KG, Gupta S, Cahalan MD (1987) Mitogen induction of ion channels in murine T lymphocytes. J Gen Physiol 89:405–420
    Google Scholar
  7. Doupnik CA, Davidson N, Lester HA (1995) The inward rectifier potassium channel family. Curr Opin Neurobiol 5:268–277
    Google Scholar
  8. Ema H, Suda T, Miura Y, Nakauchi H (1990) Colony formation of clone-sorted human hematopoietic progenitors. Blood 75:1941–1946
    Google Scholar
  9. Gallin EK, McKinney LC (1988) Patch-clamp studies in human macrophages: single-channel and whole-cell characterization of two K+ conductances. J Membr Biol 103:55–66
    Google Scholar
  10. Grinstein S, Dixon SJ (1989) Ion transport, membrane potential, and cytoplasmic pH in lymphocytes: changes during activation. Physiol Rev 69:417–481
    Google Scholar
  11. Hagiwara S, Miyazaki S, Krasne S, Ciani S (1977) Anomalous permeabilities of the egg cell membrane of a starfish in K+-T1+ mixtures. J Gen Physiol 70:269–281
    Google Scholar
  12. Hannon K, Smith II CK, Bales KR, Santerre RF (1992) Temporal and quantitative analysis of myogenic regulatory and growth factor gene expresssion in the developing mouse embryo. Dev Biol 151:137–144
    Google Scholar
  13. Hansen MB, Nielsen SE, 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
    Google Scholar
  14. Hille B (1992) Ionic channels of excitable membranes, 2nd edn. Sinauer, Sunderland, Mass., USA
    Google Scholar
  15. Ho K, Nichols CG, Lederer WJ, Lytton J, Vassilev PM, Kanazirska MV, Hebert SC (1993) Cloning and expression of an inwardly rectifying ATP-regulated potassium channel. Nature 362:31–38
    Google Scholar
  16. Horie M, Broxmeyer HE (1993) Involvement of immediateearly gene expression in the synergistic effects of steel factor in combination with granulocyte-macrophage colony-stimulating factor or interleukin-3 on proliferation of a human factordependent cell line. J Biol Chem 268:968–973
    Google Scholar
  17. Horn R, Marty A (1988) Muscarinic activation of ionic currents measured by a new whole-cell recording method. J Gen Physiol 92:145–159
    Google Scholar
  18. Huang X-Y, Morielli AD, Peralta EG (1993) Tyrosine kinasedependent suppression of a potassium channel by the G-protein-coupled ml muscarinic acetylcholine receptor. Cell 75: 1145–1156
    Google Scholar
  19. Huang Y, Rane SG (1994) Potassium channel induction by Ras/Raf signal transduction cascade. J Biol Chem 269: 31183–31189
    Google Scholar
  20. Kanakura Y, Druker B, Cannistra SA, Furukawa Y, Torimoto Y, Griffin JD (1990) Signal transduction of the human granulocyte-macrophage colony-stimulating factor and interleukin-3 receptors involves tyrosine phosphorylation of a common set of cytoplasmic proteins. Blood 76:706–715
    Google Scholar
  21. Khan IM, Coulson JM (1993) A novel method to stabilise antisense oligonucleotides against exonuclease degradation. Nucleic Acids Res 21:2957–2958
    Google Scholar
  22. Kubo Y, Baldwin TJ, Jan YN, Jan LY (1993) Primary structure and functional expression of a mouse inward rectifier potassium channel. Nature 362:127–133
    Google Scholar
  23. Lewis JP, Trobaugh FE (1964) Haematopoietic stem cells. Nature 204:589
    Google Scholar
  24. Lu L, Yang T, Markakis D, Guggino WB, Craig RW (1993) Alterations in a voltage-gated K+ current during the differentiation of ML-1 human mycloblastic leukemia cells. J Membr Biol 132:267–274
    Google Scholar
  25. Mauro TM, Isseroff RR, Lasarow R, Pappone PA (1993) Ion channels are linked to differentiation in keratinocytes. J Membr Biol 132:201–209
    Google Scholar
  26. McKinney LC, Gallin EK (1992) G-protein activators induce a potassium conductance in murine macrophages. J Membr Biol 130:265–276
    Google Scholar
  27. Merchav S, Silvian-Drachsler I, Tatarsky I, Lake M, Skottner A (1992) Comparative studies of the erythroid-potentiating effects of biosynthetic human insulin-like growth factors-I and -II. J Clin Endocrinol Metab 74:447–452
    Google Scholar
  28. Migliaccio G, Migliaccio AR, Druzin ML, Giardina PJ, Zsebo KM, Adamson JW (1991) Effects of recombinant human stem cell factor (SCF) on the growth of human progenitor cells in vitro. J Cell Physiol 148:503–509
    Google Scholar
  29. Miyajima A, Kitamura T, Harada N, Yokota T, Arai K (1992) Cytokine receptors and signal transduction. Annu Rev Immunol 10:295–331
    Google Scholar
  30. Miyazawa K, Hendrie PC, Mantel C, Wood K, Ashman LK, Broxmeyer HE (1991) Comparative analysis of signaling pathways between mast cell growth factor (c-kit ligand) and granulocyte-macrophage colony-stimulating factor in a human factor-dependent myeloid cell line involves phosphorylation of Raf-1, GTPase-activating protein and mitogen-activated protein kinase. Exp Hematol 19:1110–1123
    Google Scholar
  31. Oleson DR, DeFelice LJ, Donahoe RM (1993) A comparison of K+ channel characteristics in human T cells: perforated-patch versus whole-cell recording techniques. J Membr Biol 132: 229–241
    Google Scholar
  32. Spruce AE, Moody WJ (1992) Developmental sequence of expression of voltage-dependent currents in embryonic Xenopus laevis myocytes. Dev Biol 154:11–22
    Google Scholar
  33. Stein CA, Cheng YC (1993) Antisense oligonucleotides as therapeutic agents-is the bullet really magical? Science 261: 1004–1012
    Google Scholar
  34. von Haebler D, Stahel J, Draguhn A, Heinemann U (1993) Properties of horizontal cells transiently appearing in the rat dentate gyrus during ontogenesis. Exp Brain Res 94: 33–42
    Google Scholar
  35. Wieland SJ, Chou RH, Gong QH (1990) Macrophage-colonystimulating factor (CSF-1) modulates a differentiation-specific inward-rectifying potassium current in human leukemic (HL60) cells. J Cell Physiol 142:643–651
    Google Scholar

Download references

Author information

Authors and Affiliations

  1. Dept. Biophysics and Physiology and Haemopoiesis Unit, Bruce Rappaport Faculty of Medicine, Technion, POB 9697, 31096, Haifa, Israel
    Orian Shirihai
  2. Bruce Rappaport Faculty of Medicine, Haemopoiesis Unit, Rappaport Institute for Research in the Medical Sciences, POB 9697, 31096, Technion, Haifa, Israel
    Shoshana Merchav
  3. Dept. of Neurobiology, Weizmann Institute of Science, Rehovot, Israel
    Bernard Attali
  4. Bruce Rappaport Faculty of Medicine, Department of Biophysics and Physiology and Rappaport Institute for Research in the Medical Sciences, Bernard Katz Minerva Center for Cell Biophysics, POB 9697, 31096, Technion, Haifa, Israel
    Daniel Dagan

Authors

  1. Orian Shirihai
  2. Shoshana Merchav
  3. Bernard Attali
  4. Daniel Dagan

Rights and permissions

About this article

Cite this article

Shirihai, O., Merchav, S., Attali, B. et al. K+ channel antisense oligodeoxynucleotides inhibit cytokine-induced expansion of human hemopoietic progenitors.Pflugers Arch. 431, 632–638 (1996). https://doi.org/10.1007/BF02191913

Download citation

Key words