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Differential gene expression of bone marrow-derived CD34+ cells is associated with survival of patients suffering from myelodysplastic syndrome

International Journal of Hematology volume 89, pages 173–187 (2009)Cite this article

Abstract

One feature of the molecular pathology of myelodysplastic syndromes (MDS) is aberrant gene expression. Such aberrations may be related to patient survival, and may indicate to novel diagnostic and therapeutic targets. Therefore, we aimed at identifying aberrant gene expression that is associated with MDS and patient survival. Bone marrow-derived CD34+ hematopoietic progenitor cells from six healthy persons and 16 patients with MDS were analyzed on cDNA macroarrays comprising 1,185 genes. Thereafter, our patients were followed-up for 54 months. We found differential expression of genes that were hitherto unrecognized in the context of MDS. Differential expression of 10 genes was confirmed by quantitative real-time RT-PCR. Hierarchical cluster analysis facilitated the separation of CD34+ cells of normal donors from patients with MDS. More importantly, it also distinguished MDS-patients with short and long survival. Scrutinizing our cDNA macroarray data for genes that are associated with short survival, we found, among others, increased expression of six different genes that encode the proteasome subunits. On the other hand, the most differentially down-regulated gene was IEX-1, which encodes an anti-apoptotic protein. We confirmed its decreased expression on RNA and protein level in an independent validation set of patient samples. The presented data broadens our notion about the molecular pathology of MDS and may lend itself to better identify patients with short survival. Furthermore, our findings may help to define new molecular targets for drug development and therapeutic approaches for patients with poor prognosis.

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References

  1. Koeffler HP, Golde DW. Cellular maturation in human preleukemia. Blood. 1978;52:355–61.

    CAS  PubMed  Google Scholar 

  2. Raza A, Alvi S, Borok RZ, et al. Excessive proliferation matched by excessive apoptosis in myelodysplastic syndromes: the cause-effect relationship. Leuk Lymphoma. 1997;27:111–8.

    CAS  Article  PubMed  Google Scholar 

  3. Germing U, Hildebrandt B, Pfeilstocker M, et al. Refinement of the international prognostic scoring system (IPSS) by including LDH as an additional prognostic variable to improve risk assessment in patients with primary myelodysplastic syndromes (MDS). Leukemia. 2005;19:2223–31. doi:10.1038/sj.leu.2403963.

    CAS  Article  PubMed  Google Scholar 

  4. Nyakern M, Tazzari PL, Finelli C, et al. Frequent elevation of Akt kinase phosphorylation in blood marrow and peripheral blood mononuclear cells from high-risk myelodysplastic syndrome patients. Leukemia. 2006;20:230–8. doi:10.1038/sj.leu.2404057.

    CAS  Article  PubMed  Google Scholar 

  5. Niimi H, Harada H, Harada Y, et al. Hyperactivation of the RAS signaling pathway in myelodysplastic syndrome with AML1/RUNX1 point mutations. Leukemia. 2006;20:635–44. doi:10.1038/sj.leu.2404136.

    CAS  Article  PubMed  Google Scholar 

  6. Okamoto M, Shichishima T, Noji H, et al. High frequency of several PIG-A mutations in patients with aplastic anemia and myelodysplastic syndrome. Leukemia. 2006;20:627–34. doi:10.1038/sj.leu.2404135.

    CAS  Article  PubMed  Google Scholar 

  7. Ingram W, Lea NC, Cervera J, et al. The JAK2 V617F mutation identifies a subgroup of MDS patients with isolated deletion 5q and a proliferative bone marrow. Leukemia. 2006;20:1319–21. doi:10.1038/sj.leu.2404215.

    CAS  Article  PubMed  Google Scholar 

  8. Magnusson MK, Meade KE, Nakamura R, Barrett J, Dunbar CE. Activity of STI571 in chronic myelomonocytic leukemia with a platelet-derived growth factor beta receptor fusion oncogene. Blood. 2002;100:1088–91. doi:10.1182/blood-2002-01-0165.

    CAS  Article  PubMed  Google Scholar 

  9. Prall WC, Czibere A, Jager M, et al. Age-related transcription levels of KU70, MGST1 and BIK in CD34+ hematopoietic stem and progenitor cells. Mech Ageing Dev. 2007;128:503–10. doi:10.1016/j.mad.2007.06.008.

    CAS  Article  PubMed  Google Scholar 

  10. Steidl U, Kronenwett R, Rohr UP, et al. Gene expression profiling identifies significant differences between the molecular phenotypes of bone marrow-derived and circulating human CD34+ hematopoietic stem cells. Blood. 2002;99:2037–44. doi:10.1182/blood.V99.6.2037.

    CAS  Article  PubMed  Google Scholar 

  11. Herrler M. Use of SMART-generated cDNA for differential gene expression studies. J Mol Med. 2000;78:B23.

    CAS  PubMed  Google Scholar 

  12. Steidl U, Bork S, Schaub S, et al. Primary human CD34+ hematopoietic stem and progenitor cells express functionally active receptors of neuromediators. Blood. 2004;104:81–8. doi:10.1182/blood-2004-01-0373.

    CAS  Article  PubMed  Google Scholar 

  13. Kang HL, Benzer S, Min KT. Life extension in Drosophila by feeding a drug. Proc Natl Acad Sci USA. 2002;99:838–43. doi:10.1073/pnas.022631999.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  14. Modlich O, Prisack HB, Munnes M, Audretsch W, Bojar H. Immediate gene expression changes after the first course of neoadjuvant chemotherapy in patients with primary breast cancer disease. Clin Cancer Res. 2004;10:6418–31. doi:10.1158/1078-0432.CCR-04-1031.

    CAS  Article  PubMed  Google Scholar 

  15. Prosniak M, Hooper DC, Dietzschold B, Koprowski H. Effect of rabies virus infection on gene expression in mouse brain. Proc Natl Acad Sci USA. 2001;98:2758–63. doi:10.1073/pnas.051630298.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  16. Huang H, Colella S, Kurrer M, Yonekawa Y, Kleihues P, Ohgaki H. Gene expression profiling of low-grade diffuse astrocytomas by cDNA arrays. Cancer Res. 2000;60:6868–74.

    CAS  PubMed  Google Scholar 

  17. Beissbarth T, Fellenberg K, Brors B, et al. Processing and quality control of DNA array hybridization data. Bioinformatics. 2000;16:1014–22. doi:10.1093/bioinformatics/16.11.1014.

    CAS  Article  PubMed  Google Scholar 

  18. Quackenbush J. Computational analysis of microarray data. Nat Rev Genet. 2001;2:418–27. doi:10.1038/35076576.

    CAS  Article  PubMed  Google Scholar 

  19. Armstrong SA, Staunton JE, Silverman LB, et al. MLL translocations specify a distinct gene expression profile that distinguishes a unique leukemia. Nat Genet. 2002;30:41–7. doi:10.1038/ng765.

    CAS  Article  PubMed  Google Scholar 

  20. Golub TR, Slonim DK, Tamayo P, et al. Molecular classification of cancer: class discovery and class prediction by gene expression monitoring. Science. 1999;286:531–7. doi:10.1126/science.286.5439.531.

    CAS  Article  PubMed  Google Scholar 

  21. Spentzos D, Levine DA, Ramoni MF, et al. Gene expression signature with independent prognostic significance in epithelial ovarian cancer. J Clin Oncol. 2004;22:4700–10. doi:10.1200/JCO.2004.04.070.

    Article  PubMed  Google Scholar 

  22. Wu B, Abbott T, Fishman D, et al. Comparison of statistical methods for classification of ovarian cancer using mass spectrometry data. Bioinformatics. 2003;19:1636–43. doi:10.1093/bioinformatics/btg210.

    CAS  Article  PubMed  Google Scholar 

  23. Li C, Wong WH. Model-based analysis of oligonucleotide arrays: expression index computation and outlier detection. Proc Natl Acad Sci USA. 2001;98:31–6. doi:10.1073/pnas.011404098.

    CAS  Article  PubMed  Google Scholar 

  24. Chen G, Zeng W, Miyazato A, et al. Distinctive gene expression profiles of CD34 cells from patients with myelodysplastic syndrome characterized by specific chromosomal abnormalities. Blood. 2004;104:4210–8. doi:10.1182/blood-2004-01-0103.

    CAS  Article  PubMed  Google Scholar 

  25. Hofmann WK, de Vos S, Komor M, Hoelzer D, Wachsman W, Koeffler HP. Characterization of gene expression of CD34+ cells from normal and myelodysplastic bone marrow. Blood. 2002;100:3553–60. doi:10.1182/blood.V100.10.3553.

    CAS  Article  PubMed  Google Scholar 

  26. Mano H. DNA micro-array analysis of myelodysplastic syndrome. Leuk Lymphoma. 2006;47:9–14. doi:10.1080/10428190500264231.

    CAS  Article  PubMed  Google Scholar 

  27. Miyazato A, Ueno S, Ohmine K, et al. Identification of myelodysplastic syndrome-specific genes by DNA microarray analysis with purified hematopoietic stem cell fraction. Blood. 2001;98:422–7. doi:10.1182/blood.V98.2.422.

    CAS  Article  PubMed  Google Scholar 

  28. Pradhan A, Mijovic A, Mills K, et al. Differentially expressed genes in adult familial myelodysplastic syndromes. Leukemia. 2004;18:449–59. doi:10.1038/sj.leu.2403265.

    CAS  Article  PubMed  Google Scholar 

  29. Ueda M, Ota J, Yamashita Y, et al. DNA microarray analysis of stage progression mechanism in myelodysplastic syndrome. Br J Haematol. 2003;123:288–96. doi:10.1046/j.1365-2141.2003.04601.x.

    CAS  Article  PubMed  Google Scholar 

  30. Pellagatti A, Esoof N, Watkins F, et al. Gene expression profiling in the myelodysplastic syndromes using cDNA microarray technology. Br J Haematol. 2004;125:576–83. doi:10.1111/j.1365-2141.2004.04958.x.

    CAS  Article  PubMed  Google Scholar 

  31. Pellagatti A, Fidler C, Wainscoat JS, Boultwood J. Gene expression profiling in the myelodysplastic syndromes. Hematology. 2005;10:281–7. doi:10.1080/10245330500065680.

    CAS  Article  PubMed  Google Scholar 

  32. Steensma DP, Tefferi A. The myelodysplastic syndrome(s): a perspective and review highlighting current controversies. Leuk Res. 2003;27:95–120. doi:10.1016/S0145-2126(02)00098-X.

    Article  PubMed  Google Scholar 

  33. Allampallam K, Shetty V, Hussaini S, et al. Measurement of mRNA expression for a variety of cytokines and its receptors in bone marrows of patients with myelodysplastic syndromes. Anticancer Res. 1999;19:5323–8.

    CAS  PubMed  Google Scholar 

  34. Holmes J, Wareing C, Jacobs A, Hayes JD, Padua RA, Wolf CR. Glutathione-s-transferase pi expression in leukaemia: a comparative analysis with mdr-1 data. Br J Cancer. 1990;62:209–12.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  35. Ries C, Loher F, Zang C, Ismair MG, Petrides PE. Matrix metalloproteinase production by bone marrow mononuclear cells from normal individuals and patients with acute and chronic myeloid leukemia or myelodysplastic syndromes. Clin Cancer Res. 1999;5:1115–24.

    CAS  PubMed  Google Scholar 

  36. Gasson JC, Golde DW, Kaufman SE, et al. Molecular characterization and expression of the gene encoding human erythroid-potentiating activity. Nature. 1985;315:768–71. doi:10.1038/315768a0.

    CAS  Article  PubMed  Google Scholar 

  37. Guedez L, Stetler-Stevenson WG, Wolff L, et al. In vitro suppression of programmed cell death of B cells by tissue inhibitor of metalloproteinases-1. J Clin Invest. 1998;102:2002–10. doi:10.1172/JCI2881.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  38. Chen J, Willingham T, Margraf LR, Schreiber-Agus N, DePinho RA, Nisen PD. Effects of the MYC oncogene antagonist, MAD, on proliferation, cell cycling and the malignant phenotype of human brain tumour cells. Nat Med. 1995;1:638–43. doi:10.1038/nm0795-638.

    CAS  Article  PubMed  Google Scholar 

  39. Postel EH, Berberich SJ, Flint SJ, Ferrone CA. Human c-myc transcription factor PuF identified as nm23-H2 nucleoside diphosphate kinase, a candidate suppressor of tumor metastasis. Science. 1993;261:478–80. doi:10.1126/science.8392752.

    CAS  Article  PubMed  Google Scholar 

  40. Mitra S, Boldogh I, Izumi T, Hazra TK. Complexities of the DNA base excision repair pathway for repair of oxidative DNA damage. Environ Mol Mutagen. 2001;38:180–90. doi:10.1002/em.1070.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  41. Kane SE, Gottesman MM. The role of cathepsin L in malignant transformation. Semin Cancer Biol. 1990;1:127–36.

    CAS  PubMed  Google Scholar 

  42. Taylor GA, Thompson MJ, Lai WS, Blackshear PJ. Phosphorylation of tristetraprolin, a potential zinc finger transcription factor, by mitogen stimulation in intact cells and by mitogen-activated protein kinase in vitro. J Biol Chem. 1995;270:13341–7. doi:10.1074/jbc.270.22.13341.

    CAS  Article  PubMed  Google Scholar 

  43. Carballo E, Lai WS, Blackshear PJ. Feedback inhibition of macrophage tumor necrosis factor-alpha production by tristetraprolin. Science. 1998;281:1001–5. doi:10.1126/science.281.5379.1001.

    CAS  Article  PubMed  Google Scholar 

  44. Kitagawa M, Saito I, Kuwata T, et al. Overexpression of tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma by bone marrow cells from patients with myelodysplastic syndromes. Leukemia. 1997;11:2049–54. doi:10.1038/sj.leu.2400844.

    CAS  Article  PubMed  Google Scholar 

  45. Huang YH, Wu JY, Zhang Y, Wu MX. Synergistic and opposing regulation of the stress-responsive gene IEX-1 by p53, c-Myc, and multiple NF-kappaB/rel complexes. Oncogene. 2002;21:6819–28. doi:10.1038/sj.onc.1205854.

    CAS  Article  PubMed  Google Scholar 

  46. Rajapaksa R, Ginzton N, Rott LS, Greenberg PL. Altered oncoprotein expression and apoptosis in myelodysplastic syndrome marrow cells. Blood. 1996;88:4275–87.

    CAS  PubMed  Google Scholar 

  47. Kaeferstein A, Krug U, Tiesmeier J, et al. The emergence of a C/EBPalpha mutation in the clonal evolution of MDS towards secondary AML. Leukemia. 2003;17:343–9. doi:10.1038/sj.leu.2402805.

    CAS  Article  PubMed  Google Scholar 

  48. Fuhler GM, Drayer AL, Vellenga E. Decreased phosphorylation of protein kinase B and extracellular signal-regulated kinase in neutrophils from patients with myelodysplasia. Blood. 2003;101:1172–80. doi:10.1182/blood.V101.3.1172.

    CAS  Article  PubMed  Google Scholar 

  49. Letourneux C, Rocher G, Porteu F. B56-containing PP2A dephosphorylate ERK and their activity is controlled by the early gene IEX-1 and ERK. EMBO J. 2006;25:727–38. doi:10.1038/sj.emboj.7600980.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  50. Kruse ML, Arlt A, Sieke A, et al. Immediate early gene X1 (IEX-1) is organized in subnuclear structures and partially co-localizes with promyelocytic leukemia protein in HeLa cells. J Biol Chem. 2005;280:24849–56. doi:10.1074/jbc.M501571200.

    CAS  Article  PubMed  Google Scholar 

  51. Kumar R, Lutz W, Frank E, Im HJ. Immediate early gene X-1 interacts with proteins that modulate apoptosis. Biochem Biophys Res Commun. 2004;323:1293–8. doi:10.1016/j.bbrc.2004.09.006.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  52. Wang X, Studzinski GP. Antiapoptotic action of 1, 25-dihydroxyvitamin D3 is associated with increased mitochondrial MCL-1 and RAF-1 proteins and reduced release of cytochrome c. Exp Cell Res. 1997;235:210–7. doi:10.1006/excr.1997.3667.

    CAS  Article  PubMed  Google Scholar 

  53. Parker JE, Mufti GJ. Excessive apoptosis in low risk myelodysplastic syndromes (MDS). Leuk Lymphoma. 2000;40:1–24. doi:10.3109/10428190009054877.

    CAS  Article  PubMed  Google Scholar 

  54. Wu MX, Ao Z, Prasad KV, Wu R, Schlossman SF. IEX-1L, an apoptosis inhibitor involved in NF-kappaB-mediated cell survival. Science. 1998;281:998–1001. doi:10.1126/science.281.5379.998.

    CAS  Article  PubMed  Google Scholar 

  55. Czibere A, Prall WC, Zerbini LF, et al. The nonsteroidal anti-inflammatory drug exisulind selectively induces apoptosis via JNK in secondary acute myeloid leukemia after myelodysplastic syndrome. Cell Cycle. 2005;4:812–7.

    CAS  Article  PubMed  Google Scholar 

  56. Czibere A, Prall WC, Zerbini LF, et al. Exisulind induces apoptosis in advanced myelodysplastic syndrome (MDS) and acute myeloid leukaemia/MDS. Br J Haematol. 2006;135:355–7. doi:10.1111/j.1365-2141.2006.06298.x.

    CAS  Article  PubMed  Google Scholar 

  57. List A, Beran M, DiPersio J, et al. Opportunities for Trisenox (arsenic trioxide) in the treatment of myelodysplastic syndromes. Leukemia. 2003;17:1499–507. doi:10.1038/sj.leu.2403021.

    CAS  Article  PubMed  Google Scholar 

  58. Arlt A, Minkenberg J, Kocs B, et al. The expression of immediate early gene X-1 (IEX-1) is differentially induced by retinoic acids in NB4 and KG1 cells: possible implication in the distinct phenotype of retinoic acid-responsive and -resistant leukemic cells. Leukemia. 2004;18:1646–55. doi:10.1038/sj.leu.2403481.

    CAS  Article  PubMed  Google Scholar 

  59. Semlali A, Oliva J, Badia E, Pons M, Duchesne MJ. Immediate early gene X-1 (IEX-1), a hydroxytamoxifen regulated gene with increased stimulation in MCF-7 derived resistant breast cancer cells. J Steroid Biochem Mol Biol. 2004;88:247–59. doi:10.1016/j.jsbmb.2003.12.005.

    CAS  Article  PubMed  Google Scholar 

  60. Braun T, Carvalho G, Coquelle A, Vozenin MC, Lepelley P, Hirsch F, et al. NF-kappaB constitutes a potential therapeutic target in high-risk myelodysplastic syndrome. Blood. 2006;107:1156–65. doi:10.1182/blood-2005-05-1989.

    CAS  Article  PubMed  Google Scholar 

  61. Terpos E, Verrou E, Banti A, Kaloutsi V, Lazaridou A, Zervas K. Bortezomib is an effective agent for MDS/MPD syndrome with 5q- anomaly and thrombocytosis. Leuk Res. 2007;31:559–62. doi:10.1016/j.leukres.2006.05.018.

    CAS  Article  PubMed  Google Scholar 

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Acknowledgments

We are indebted to our colleagues Christoph Maintz, Ulrich Grabenhorst, and Christoph Losem for their support, and to Bärbel Junge, Maria Wolf, Anke Boekmann, Elke Rosenbaum-König, and Hildegard Gaussmann for cell separations. Further, we are grateful to Annette Schreiber, and Claudia Aivado for the visual assessment of arrays. Finally, I thank Else-Marie Meyer for her inspiring lessons in molecular genetics, and Carlo Aul for his cordial support. This work was supported by the Bundesministerium für Bildung und Forschung (BMBF), Kompetenznetz “Akute und Chronische Leukämien” (No. TP16), and the Leukämie-Liga Düsseldorf, e.V.

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Affiliations

  1. Department of Hematology, Oncology, and Clinical Immunology, Heinrich-Heine-University, Düsseldorf, Germany

    Wolf C. Prall, Akos Czibere, Andrea Kuendgen, Astrid Rong, Ulrich Germing, Norbert Gattermann, Rainer Haas & Manuel Aivado

  2. Beth Israel Deaconess Medical Center and Harvard Medical School, Harvard Institutes of Medicine, Boston, MA, USA

    Akos Czibere, Franck Grall, Dimitrios Spentzos, Hasan Otu, Towia A. Libermann & Manuel Aivado

  3. Department of Cell Biology, Albert Einstein Cancer Center, Albert Einstein College of Medicine, Bronx, NY, USA

    Ulrich Steidl

  4. Department of Hematology, Oncology, and Clinical Immunology, St Johannes Hospital, Duisburg, Germany

    Aristoteles Achilles Nikolaus Giagounidis

  5. Experimental Surgery and Regenerative Medicine, Department of Surgery, Ludwig-Maximilians-University, Nussbaumstraße 20, 80336, Munich, Germany

    Wolf C. Prall

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  1. Wolf C. Prall
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  2. Akos Czibere
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Correspondence to Wolf C. Prall.

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Prall, W.C., Czibere, A., Grall, F. et al. Differential gene expression of bone marrow-derived CD34+ cells is associated with survival of patients suffering from myelodysplastic syndrome. Int J Hematol 89, 173–187 (2009). https://doi.org/10.1007/s12185-008-0242-9

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  • DOI: https://doi.org/10.1007/s12185-008-0242-9

Keywords

  • cDNA array
  • Gene expression
  • MDS
  • Survival
  • IEX-1