Biometals

, Volume 17, Issue 4, pp 443–450 | Cite as

Effects of AZT on cellular iron homeostasis

  • Argante Bozzi
  • Fabrizia Brisdelli
  • Anna M. D'Alessandro
  • Gabriele D'Andrea
  • Anna R. Lizzi
  • Andrea C. Rinaldi
  • Arduino Oratore

Abstract

3′-azido-3′-deoxythymidine (AZT), the first chemotherapeutic drug approved by FDA for treatment of HIV-infected patients and still used in combination therapy, has been shown to induce, upon prolonged exposure, severe bone marrow toxicity manifested as anemia, neutropenia and siderosis. These toxic effects are caused by inhibition of heme synthesis and, as a consequence, transferrin receptor (TfR) number appears increased and so iron taken up by cells. Since iron overload can promote the frequency and severity of many infections, siderosis is viewed as a further burden for AIDS patients. We have previously demonstrated that AZT-treated K562 cells showed an increase of the number of TfRs located on the surface of the plasma membrane without affecting their biosynthesis, but slowing down their endocytotic pathway. In spite of the higher number of receptors on the plasma-membrane of AZT-treated cells, intracellular accumulation of iron showed a similar level in control and in drug-exposed cells. The chelating ability of AZT and of its phosphorylated derivatives, both in an acellular system and in K562 cells, was also checked. The results demonstrated that AZT and AZTMP were uneffective as iron chelators, while AZTTP displayed a significant capacity to remove iron from transferrin (Tf). Our results suggest that AZT may be not directly involved in the iron overloading observed upon its prolonged use in AIDS therapy. The iron accumulation found in these patients is instead caused by other unknown mechanisms that need further studies to be clarified.

3′-azido-3′-deoxythymidine (AZT) iron transferrin receptor (TfR) glycosylation chelatable 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Abraham NG, Lutton JD, Levere RD. 1985 Heme metabolism and erythropoiesis in abnormal iron states: role of delta-aminolevulinic acid synthase and heme oxygenase. Exp Hematol 13, 838-843.Google Scholar
  2. Avramis VI, Kwock R, Solorzano MM, Gomperts E. 1993 Evidence of in vitro development of drug resistance to azido-thymidine in T-lymphocytic leukemia cell lines (Jurkat E61/AZT-100) and in pediatric patients with HIV-infection. J AIDS 6, 1287-1296.Google Scholar
  3. Balzarini J, Pauwels R, Baba M, et al. 1988 The in vitro and in vivo antiretrovirus activity, and intracellular metabolism of 3´-azido-2'´,3´-dideoxythymidine and 2´,3´-dideoxycithydine are highly dependent on the cell species. Biochem Pharmacol 37, 897-903.Google Scholar
  4. Boelaert JR, Weinberg GA, Weinberg ED. 1996 Altered iron metabolism in HIV infection: Mechanism, possible consequences, and proposals for management. Infect Agents Dis 5, 36-46.Google Scholar
  5. Breuer W, Epsztejn S, Cabantchik ZI. 1995 Iron acquired from transferrin by K562 cells is delivered into a cytoplasmic pool of chelatable iron (II). J Biol Chem 270, 24209-24215.Google Scholar
  6. Brever W, Epsztejn S, Cabantchik ZI. 1996 Dynamics of the cytosolic chelatable iron pool of K562 cells. FEBS Lett 382, 304-308.Google Scholar
  7. Bridges EG, Trentesaux C, Lahlil R, Spiga MG, Jeannesson P, Sommadossi JP. 1996 3´-azido-3´-deoxythymidine inhibits erythroid-specific transcription factors in human erythroid K562 leukemia cells. Eur J Haematol 56, 62-67.Google Scholar
  8. Casey JL, Hentze MW, Koeller DM, et al. 1988 Iron-responsive elements: regulatory RNA sequences that control mRNA levels and translation. Science 240, 924-928.Google Scholar
  9. Chaudary PM and Roninson IB. 1993 Induction of multidrug resistance in human cells by transient exposure to different chemotherapeutic drugs. J Natl Cancer Inst 85, 632-639.Google Scholar
  10. Chen MS, Woods KL, Prusoff WH. 1984 Molecular basis of the antineoplastic activity of 3´-amino-3´-deoxythymidine. Mol Pharmacol 25, 441-445.Google Scholar
  11. Cheng YC, Dutschman GE, Bastow KF, Sarngadharan MG, Ting RY. 1987 Human immunodeficiency virus reverse transcriptase: general properties and its interactions with nucleoside triphosphate analogs. J Biol Chem 262, 2187-2189.Google Scholar
  12. Chiba T, Ikawa Y, Todokoro K. 1991 GATA-1 transactivates erythropoietin receptor gene, and erythropoietin receptor-mediated signals enhance GATA-1 gene expression. Nucleic Acids Res 19, 3843-3848.Google Scholar
  13. Childs S and Ling V. 1994 The MDR superfamily of genes and its biological implications. In: De Vita VT, Hellmann S, Rosenberg SA eds. Important Advances in Oncology. JB Lippincott Co, Philadelphia: 21-36.Google Scholar
  14. Chow FPR, Sutton PA, Hamburger AW. 1991 Sensitivity of erythroid progenitor colonies to erythropoietin in azidothymidine treated immunodeficient mice. Br J Haematol 77, 139-144.Google Scholar
  15. Cretton EM, Xie MY, Bevan RJ, Goudgaon NM, Schinazi RF, Sommadossi JP. 1991 Catabolism of 3´-azido-3´-deoxythymidine in hepatocytes and liver microsomes, with evidence of formation of 3´-amino-3´-deoxythymidine, a highly toxic catabolite for human bone marrow cells. Mol Pharmacol 39, 258-266.Google Scholar
  16. D'Alessandro AM, D'Andrea G, Di Ciccio L, et al. 1999 3´-azido-3´-deoxythymidine reduces the rate of transferrin receptor endocytosis in K562 cells. Biochim Biophys Acta 1450, 232-241.Google Scholar
  17. D'Alessandro AM, Rinaldi AC, D'Andrea G, et al. 2000 Evidences that zidovudine (AZT) could not be directly responsible for iron overload in AZT-treated patients: an in vitro study. Clin Chim Acta 300, 119-130.Google Scholar
  18. D'Andrea G, D'Alessandro AM, Brisdelli F, et al. 1999 Effect of AZT on human transferrin receptor sislylation process in K562 cells. In: World Congress on Iron Metabolism. Bioiron 99, Sorrento, Italy.Google Scholar
  19. D'Andrea G, Di Ciccio L, Brisdelli F, D'Alessandro AM, Bozzi A, Oratore A. 2001 Purification and partial characterization of an α-2,8-sialyltransferase from human erythroleukemia K562 cells. Prep Biochem Biotechnol 31, 355-368.Google Scholar
  20. D'Andrea G, Lizzi AR, Brisdelli F, D'Alessandro AM, Bozzi A, Oratore A. 2003 Protein glycans alteration and a different distribution of some enzymatic activities involved in the glycan processing are found in AZT-treated K562 cells. Mol Cell Biochem 252, 45-51.Google Scholar
  21. Dalakas MC, Illa I, Pezeshkpour GH, Laukaitis JP, Cohen B, Griffin JL. 1990 Mitochondrial myopathy caused by long-term zidovudine therapy. N Engl J Med 322, 1098-1105.Google Scholar
  22. Di Vito M, Bozzi A, Ferretti A, et al. 1997 Intracellular metabolism of 3´-azido-3´-deoxythymidine (AZT): A Nuclear Magnetic Resonance study on T-lymphoblastoid cell lines with different resistance to AZT. Biochem Pharmacol 54, 979-990.Google Scholar
  23. Dianzani F, Antonelli G, Turriziani O, et al. 1994 Zidovudine induces the expression of cellular resistance affecting its antiviral activity. AIDS Res Hum Retrov 10, 1471-1478.Google Scholar
  24. Domin BA, Mahony WB, Zimmerman TP. 1988 Purine nucleo-base transport in human erythrocytes: reinvestigation with a novel 'inhibitor-stop' assay. J Biol Chem 263, 9276-9284.Google Scholar
  25. Furman PA, Fyfe JA, St. Clair MH. 1986 Phosphorylation of 3´-azido-3´-deoxythymidine and selective interaction of the 5´-triphosphate with human immunodeficiency virus reverse transcriptase. Proc Natl Acad Sci USA 83, 8333-8337.Google Scholar
  26. Germann UA, Pastan I, Gottesman MM. 1993 P-glycoproteins: mediators of multidrug resistance. Semin Cell Biol 4, 63-76.Google Scholar
  27. Gill PS, Rarick M, Brynes RK, Causey D, Loureiro C, Levine AM. 1987 Azidothymidine associated with bone marrow failure in the acquired immunodeficiency syndrome (AIDS). Ann Intern Med 107, 502-505.Google Scholar
  28. Gogu SR, Malter JS, Agrawal KC. 1992 Zidovudine-induced blockade of the expression and function of the erythropoietin receptor. Biochem Pharmacol 44, 1009-1012.Google Scholar
  29. Goldin RD, Wilkins M, Dourakis S, Parkin J, Lindley R. 1993 Iron overload in multiply transfused patients who are HIV seropositive. J Clin Pathol 46, 1036-1038.Google Scholar
  30. Good SS, Koble CS, Crouch R, Johnson RL, Rideout JL, deMiranda P. 1990 Isolation and characterization of an ether glucuronide of zidovudine, a major metabolite in monkeys and humans. Drug Metab Dispos 18, 321-326.Google Scholar
  31. Gordeuk V, Delanghe J, Langlois M, Boelaert J. 2001 Iron status and the outcome of HIV infection: an overview. J Clin Virol 20, 111-115.Google Scholar
  32. Groopman J. 1997 Impact of transfusion on viral load in human immunodeficiency virus infection. Semin Hematol 34, 27-33.Google Scholar
  33. Hall ET, Yan JP, Melançon P, Kuchta RD. 1994 3´-axido-3´-deoxythymidine potentially inhibits protein glycosylation. J Biol Chem 269, 14355-14358.Google Scholar
  34. Henry DH, Beall GN, Benson CA, et al. 1992 Recombinant human erythropoietin in the treatment of anemia associated with HIV infection and zidovudine therapy. Overview of four clinical trials. Ann Intern Med 117, 739-748.Google Scholar
  35. Hermans P and Clumeck N. 1995 Kaposi's sarcoma in patients infected with human virus (HIV): an overview. Cell Mol Biol 3, 357-364.Google Scholar
  36. Hershko C, Link G, Cabantchik ZI. 1998 Pathophysiology of iron overload. Ann New York Acad Sci 850, 191-201.Google Scholar
  37. Jacopetta BJ, Morgan EH, Yeoh GCT. 1982 Transferrin receptors and iron uptake during erythroid development. Biochim Biophys Acta 687, 204-210.Google Scholar
  38. Jandl JH and Katz JH. 1963 The plasma-to-cell cycle of transferrin. J Clin Invest 42, 314-326.Google Scholar
  39. Kartner N, Riordan JR, Ling V. 1983 Cell surface P-glycoprotein associated with multidrug resistance in mammalian cell lines. Science 221, 1058-1088.Google Scholar
  40. Kedar PS, Abbots J, Kovacs T, Lesiak K, Torrence W, Wilson SH. 1990 Mechanism of HIV reverse transcriptase: enzyme-primer interaction as revealed through studies of a dNTP analogue, 3´-azidothymidine-5´-triphosphate. Biochemistry 29, 3603-3611.Google Scholar
  41. Kornfeld S and Mellman I. 1989 The biogenesis of lysosomes. Annu Rev Cell Biol 5, 483-525.Google Scholar
  42. Kreutzer KA and Rockstrom JK. 1997 Pathogenesis and pathophysiology of anemia in HIV infection. Ann Hematol 75, 179-187.Google Scholar
  43. Langtry HD and Campoli-Richards DM. 1989 Zidovudine. A review of its pharmacodynamic and pharmacokinetic properties, and therapeutic efficacy. Drugs 37, 408-450.Google Scholar
  44. Lin JJ, Patino M, Gaffield L, Walden WE, Smith A, Thach RE. 1991 Crosslinking of hemin to a specific site on the 90 KD ferritin repressor protein. Proc Natl Acad Sci USA88, 6068-6071.Google Scholar
  45. Lindley R, Parkin J, Dourakis S, Goldin R. 1989 Iron deposition in liver in zidovudine-related transfusion-dependent anaemia. Lancet 11, 681.Google Scholar
  46. Lucarelli M, Palitti M, Carotti D, et al. 1996 AZT-induced hypermethylation of human thymidine kinase gene in the absence of total DNA hypermethylation. FEBS Lett 396, 323-326.Google Scholar
  47. Ludwig H and Strasser K. 2001 Symptomatology of anemia. Semin Oncol 28, 7-14.Google Scholar
  48. Lutton JD, Mathew A, Levere RD, Abraham NG. 1990 Role of heme metabolism in AZT-induced bone marrow toxicity. Am J Hematol 35, 1-5.Google Scholar
  49. Martin DK, Tsai SF, Orkin SH. 1989 Increased γ-globin expression in a nondeletion HPFH mediated by an erythroid-specific DNA-binding factor. Nature 338, 435-438.Google Scholar
  50. Melefors Ö and Hentze MW. 1993 Iron regulatory factor-the conductor of cellular iron regulation. Blood Rev 7, 251-258.Google Scholar
  51. Müllner EW, Neupert B, Kühn LC. 1989 A specific mRNA binding factor regulates the iron-dependent stability of cytoplasmic transferrin receptor mRNA. Cell 58, 373-382.Google Scholar
  52. Nicolis S, Ronchi A, Malgaretti N, Mantovani R, Giglioni B, Ottolenghi S. 1989 Increased erythroid-specific expression of a mutated HPFH γ-globin promoter requires the erythroid factor GATA-1. Nucleic Acids Res 17, 5509-5516.Google Scholar
  53. Nicolis S, Bertini C, Ronchi A, et al. 1991 An erythroid specific enhancer upstream to the gene encoding the cell-type specific transcription factor GATA-1. Nucleic Acids Res 19, 5285-5291.Google Scholar
  54. Nyce J, Leonard S, Canupp D, Schulz S, Wong S. 1993 Epigenetic mechanisms of drug resistance: drug-induced DNA hypermethylation and drug resistance. Proc Natl Acad Sci USA 90, 2960-2964.Google Scholar
  55. Owen D and Kühn LC. 1987 Noncoding 3´ sequences of the transferrin receptor gene are required for mRNA regulation by iron. EMBO J 6, 1287-1293.Google Scholar
  56. Pelicci PG, Tabilio A, Thomopoulos P. 1982 Hemin regulates the expression of transferrin receptors in human hematopoietic cell lines. FEBS Lett 145, 350-354.Google Scholar
  57. Pelosi-Testa E, Testa U, Samoggia G, Salvo A, Camagna A, Peschle C. 1986 Expression of transferrin receptors in human erythroleukemia lines: Regulation in the plateau and exponential phase of growth. Cancer Res 46, 5330-5334.Google Scholar
  58. Pollack S and Weaver J. 1993 Azidothymidine (AZT)-induced siderosis. Am J Hematol 43, 230-233.Google Scholar
  59. Resetar A and Spector T. 1989 Glucuronidation of 3´-azido-3´-deoxythymidine: human and rat enzyme specificity. Biochem Pharmacol 38, 1389-1393.Google Scholar
  60. Richmann DD, Fischl MA, Grieco MH, et al. 1987 The toxicity of azidothymidine (AZT) in the treatment of patients with AIDS and AIDS-related complex: a double-blind placebo-controlled trial. N Engl J Med 317, 192-197.Google Scholar
  61. Ryan TR and Aust SD. 1992 The role of iron in oxygen-mediated toxicities. Crit Rev Toxicol 22, 119-141.Google Scholar
  62. Salhi Y, Costagliola D, Rebulla P, et al. 1998 Serum ferritin, desferrioxamine, and evolution of HIV-1 infection in thalassemic patients. J AIDS Hum Retrovirol 18, 473-478.Google Scholar
  63. Schinkel AH and Borst P. 1991 Multidrug resistance mediated by P-glycoproteins. Semin Cancer Biol 2, 213-226.Google Scholar
  64. Schonhorn JE, Akompong T, Wessling-Resnick M. 1995 Mechanism of transferrin receptor down-regulation in K562 cells in response to protein kinase C activation. J Biol Chem 270, 3698-3705.Google Scholar
  65. Seligman PA, Schleicher RB, Allen RH. 1979 Isolation and characterization of the transferrin receptor from human placenta. J Biol Chem 254, 9943-9946.Google Scholar
  66. Signoretti C, Romagnoli G, Turriziani O, Antonelli G, Dianzani F, Cianfriglia M. 1997 Induction of the multidrug-transporter Pglycoprotein by AZT treatment in tumor cell lines. J Exp Clin Cancer Res 16, 29-32.Google Scholar
  67. Sullivan PS, Hanson DL, Chu SY, Jones JL, Ward JW. 1998 Epidemiology of anaemia in HIV-infected persons: Results from the multistate adult and adolescent spectrum of HIV disease surveillance project. Blood 91, 301-308.Google Scholar
  68. Sussman MS and Bulkley GB. 1990 Oxygen-derived free radicals in reperfusion injury. Methods Enzymol 186, 711-722.Google Scholar
  69. Tornevik Y, Ullman B, Balzarini J, Wahren B, Eriksson S. 1995 cytotoxicity of 3´-azido-3´-deoxythymidine correlates with 3´-azidothymidine-5´-monophosphate (AZTMP) levels, whereas anti-human immunodeficiency virus (HIV) activity correlated with 3´-azidothymidine-5´-triphosphate (AZTTP) levels in cultured CEM T-lymphoblastoid cells. Biochem Pharmacol 49, 829-837.Google Scholar
  70. Trowbridge IS and Omary MB. 1981 Human cell surface glycoprotein related to cell proliferation is the receptor for transferrin. Proc Natl Acad Sci USA 78, 3039-3043.Google Scholar
  71. Volberding P. 2000 Consensus statement: anemia in HIV infection-Current trends, treatment options and practice strategies. Anemia in HIV Working Group. Clin Ther 22, 1004-1020.Google Scholar
  72. Waquar MA, Evans MJ, Manly KF, Hughes RG, Huberman JA. 1984 Effects of 2´,3´-dideoxynucleosides on mammalian cells and viruses. J Cell Physiol 121, 402-408.Google Scholar
  73. Ward JH, Jordan I, Kushner JP, Kaplan J. 1984 Heme regulation of HeLa cell transferrin receptor number. J Biol Chem 259, 13235-13240.Google Scholar
  74. Weaver J and Pollack S. 1989 Low-Mr iron isolated from guinea pig reticulocytes as AMP-Fe and ATP-Fe complexes. Biochem J 261, 787-792.Google Scholar
  75. Weaver J, Zhan H, Pollack S. 1990 Mitochondria have Fe (III) receptors. Biochem J 265, 415-419.Google Scholar
  76. Weider DA and Sommadossi JP. 1990 3´-azido-3´-deoxythymidine inhibits globin gene transcription in butyric acid-induced K562 human leukemia cells. Mol Pharmacol 38, 397-804.Google Scholar
  77. Weidner DA and Sommadossi JP. 1991 Comparative effects of 3´-azido-3´-deoxythymidine and its metabolite, 3´-amino-3´-deoxythymidine, on hemoglobin synthesis in K562 human leukemia cells. Mol Pharmacol 41, 252-258.Google Scholar
  78. Wu S, Liu X, Solorzano MM, Kwock R, Avramis VI. 1995 Development of zidovudine (AZT) resistance in Jurkat T cells is associated with decreased expression of the thymidine kinase (TK) gene and hypermethylation of the 5´ end of human TK gene. J AIDS Hum Retrovirol 8, 1-9.Google Scholar
  79. Yan JP, Ilsley D, Frohlick C, et al. 1995 Azidothymidine (Zidovudine) inhibits glycosylation and dramatically alters glycosphingolipid synthesis in whole cells at clinically relevant concentrations. J Biol Chem 270, 22836-22841.Google Scholar
  80. Yarchoan R, Mitsuya H, Myers CE, Broder S. 1989 Clinical pharmacology of 3´-azido-2´,3´-dideoxythymidine (zidovudine) and related dideoxynucleosides. N Engl J Med 321, 726-738.Google Scholar
  81. Young SP and Bomford A. 1994 Iterative endocytosis of transferrin by K562 cells. Biochem J 298, 165-170.Google Scholar
  82. Yusa K, Oh-hara T, Yamazaki A, Tsukahara S, Satoh W, Tsuruo T. 1990 Cross-resistance to anti-HIV nucleoside analogues in multidrug-resistant human cell. Biochem Biophys Res Commun 169, 986-990.Google Scholar
  83. Zimmerman TP, Mahony WB, Prus KL. 1987 3´-azido-3´-deoxythymidine: an unusual nucleoside analog that permeates the membrane of human erythrocytes and lymphocytes by nonfacilitated diffusion. J Biol Chem 262, 5748-5754.Google Scholar

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Argante Bozzi
    • 1
  • Fabrizia Brisdelli
    • 1
  • Anna M. D'Alessandro
    • 1
  • Gabriele D'Andrea
    • 1
  • Anna R. Lizzi
    • 1
  • Andrea C. Rinaldi
    • 1
  • Arduino Oratore
    • 1
  1. 1.Department of Biomedical Sciences and TechnologiesUniversity of L'AquilaL'AquilaItaly

Personalised recommendations