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Molecular Medicine

, Volume 19, Issue 1, pp 115–123 | Cite as

Differential microRNA Profiles and Their Functional Implications in Different Immunogenetic Subsets of Chronic Lymphocytic Leukemia

  • Nikos Papakonstantinou
  • Stavroula Ntoufa
  • Elisavet Chartomatsidou
  • Giorgio Papadopoulos
  • Artemis Hatzigeorgiou
  • Achiles Anagnostopoulos
  • Katerina Chlichlia
  • Paolo Ghia
  • Marta Muzio
  • Chrysoula Belessi
  • Kostas Stamatopoulos
Research Article

Abstract

Critical processes of B-cell physiology, including immune signaling through the B-cell receptor (BcR) and/or Toll-like receptors (TLRs), are targeted by microRNAs. With this in mind and also given the important role of BcR and TLR signaling and microRNAs in chronic lymphocytic leukemia (CLL), we investigated whether microRNAs could be implicated in shaping the behavior of CLL clones with distinct BcR and TLR molecular and functional profiles. To this end, we examined 79 CLL cases for the expression of 33 microRNAs, selected on the following criteria: (a) deregulated in CLL versus normal B-cells; (b) differentially expressed in CLL subgroups with distinct clinicobiological features; and, (c) if meeting (a) + (b), having predicted targets in the immune signaling pathways. Significant upregulation of miR-150, miR-29c, miR-143 and miR-223 and downregulation of miR-15a was found in mutated versus unmutated CLL, with miR-15a showing the highest fold difference. Comparison of two major subsets with distinct stereotyped BcRs and signaling signatures, namely subset 1 [IGHV1/5/7-IGKV1(D)-39, unmutated, bad prognosis] versus subset 4 [IGHV4-34/IGKV2-30, mutated, good prognosis] revealed differences in the expression of miR-150, miR-29b, miR-29c and miR-101, all down-regulated in subset 1. We were also able to link these distinct microRNA profiles with cellular phenotypes, importantly showing that, in subset 1, miR-101 downregulation is associated with overexpression of the enhancer of zeste homolog 2 (EZH2) protein, which has been associated with clinical aggressiveness in other B-cell lymphomas. In conclusion, specific miRNAs differentially expressed among CLL subgroups with distinct BcR and/or TLR signaling may modulate the biological and clinical behavior of the CLL clones.

Notes

Acknowledgments

This project was supported by the ENosAI project (code 09SYN-13-880) and cofunded by the European Union (EU) and the Hellenic General Secretariat for Research and Technology; Cariplo Foundation (Milan, Italy); and the Program Molecular Clinical Oncology-5 per mille number 9965, Associazione Italiana per la Ricerca sul Cancro (Italy).

Supplementary material

10020_2013_1901115_MOESM1_ESM.pdf (1.6 mb)
Supplementary material, approximately 1.59 MB.

References

  1. 1.
    Rozman C, Montserrat E. (1995) Chronic lymphocytic leukemia. N. Engl. J. Med. 333:1052–7.CrossRefPubMedGoogle Scholar
  2. 2.
    Ghia P, Chiorazzi N, Stamatopoulos K. (2008) Microenvironmental influences in chronic lymphocytic leukaemia: the role of antigen stimulation. J. Intern. Med. 264:549–62.CrossRefPubMedGoogle Scholar
  3. 3.
    Fais F, et al. (1998) Chronic lymphocytic leukemia B cells express restricted sets of mutated and unmutated antigen receptors. J. Clin. Invest. 102:1515–25.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Damle RN, etal. (1999) Ig V gene mutation status and CD38 expression as novel prognostic indicators in chronic lymphocytic leukemia. Blood. 94:1840–7.Google Scholar
  5. 5.
    Hamblin TJ, et al. (1999) Unmutated Ig V(H) genes are associated with a more aggressive form of chronic lymphocytic leukemia. Blood. 94:1848–54.Google Scholar
  6. 6.
    Agathangelidis A, et al. (2012) Stereotyped B-cell receptors in one-third of chronic lymphocytic leukemia: a molecular classification with implications for targeted therapies. Blood. 119:4467–75.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Messmer BT, et al. (2004) Multiple distinct sets of stereotyped antigen receptors indicate a role for antigen in promoting chronic lymphocytic leukemia. J. Exp. Med. 200:519–25.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Stamatopoulos K, et al. (2007) Over 20% of patients with chronic lymphocytic leukemia carry stereotyped receptors: Pathogenetic implications and clinical correlations. Blood. 109:259–70.CrossRefPubMedGoogle Scholar
  9. 9.
    Chiorazzi N, Ferrarini M. (2011) Cellular origin(s) of chronic lymphocytic leukemia: cautionary notes and additional considerations and possibilities. Blood. 117:1781–91.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Bourke E, et al. (2003) The Toll-like receptor repertoire of human B lymphocytes: inducible and selective expression of TLR9 and TLR10 in normal and transformed cells. Blood. 102:956–63.CrossRefPubMedGoogle Scholar
  11. 11.
    Akira S, Takeda K, Kaisho T. (2001) Toll-like receptors: critical proteins linking innate and acquired immunity. Nat. Immunol. 2:675–80.CrossRefPubMedGoogle Scholar
  12. 12.
    Rawlings DJ, Schwartz MA, Jackson SW, MeyerBahlburg A. (2012) Integration of B cell responses through Toll-like receptors and antigen receptors. Nat. Rev. Immunol. 12:282–94.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Lau CM, et al. (2005) RNA-associated autoantigens activate B cells by combined B cell antigen receptor/Toll-like receptor 7 engagement. J. Exp. Med. 202:1171–7.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Arvaniti E, et al. (2011) Toll-like receptor signaling pathway in chronic lymphocytic leukemia: distinct gene expression profiles of potential pathogenic significance in specific subsets of patients. Haematologica. 96:1644–52.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Ntoufa S, et al. (2012) Distinct innate immunity pathways to activation and tolerance in subgroups of chronic lymphocytic leukemia with distinct immunoglobulin receptors. Mol. Med. 18:1281–91.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Longo PG, et al. (2007) The Akt signaling pathway determines the different proliferative capacity of chronic lymphocytic leukemia B-cells from patients with progressive and stable disease. Leukemia. 21:110–20.CrossRefPubMedGoogle Scholar
  17. 17.
    Bartel DP. (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 116:281–97.CrossRefGoogle Scholar
  18. 18.
    Huntzinger E, Izaurralde E. (2011) Gene silencing by microRNAs: contributions of translational repression and mRNA decay. Nat. Rev. Genet. 12:99–110.CrossRefPubMedGoogle Scholar
  19. 19.
    O’Connell RM, Rao DS, Chaudhuri AA, Baltimore D. (2011) Physiological and pathological roles for microRNAs in the immune system. Nat. Rev. Immunol. 10:111–22.CrossRefGoogle Scholar
  20. 20.
    O’Neill LA, Sheedy FJ, McCoy CE. (2011) MicroRNAs: the fine-tuners of Toll-like receptor signalling. Nat. Rev. Immunol. 11:163–75.CrossRefPubMedGoogle Scholar
  21. 21.
    Iorio MV, Piovan C, Croce CM. (2010) Interplay between microRNAs and the epigenetic machinery: an intricate network. Biochim. Biophys. Acta. 1799:694–701.CrossRefPubMedGoogle Scholar
  22. 22.
    Calin GA, et al. (2002) Frequent deletions and down-regulation of micro-RNA genes miR15 and miR16 at 13q14 in chronic lymphocytic leukemia. Proc. Natl. Acad. Sci. U. S. A. 99:15524–9.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Calin GA, et al. (2004) MicroRNA profiling reveals distinct signatures in B cell chronic lymphocytic leukemias. Proc. Natl. Acad. Sci. U. S. A. 101:11755–60.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Fulci V, et al. (2007) Quantitative technologies establish a novel microRNA profile of chronic lymphocytic leukemia. Blood. 109:4944–51.CrossRefPubMedGoogle Scholar
  25. 25.
    Marton S, et al. (2008) Small RNAs analysis in CLL reveals a deregulation of miRNA expression and novel miRNA candidates of putative relevance in CLL pathogenesis. Leukemia. 22:330–8.CrossRefPubMedGoogle Scholar
  26. 26.
    Pallasch CP, et al. (2009) miRNA deregulation by epigenetic silencing disrupts suppression of the oncogene PLAG1 in chronic lymphocytic leukemia. Blood. 114:3255–64.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Stamatopoulos B, et al. (2009) microRNA-29c and microRNA-223 down-regulation has in vivo significance in chronic lymphocytic leukemia and improves disease risk stratification. Blood. 113:5237–45.CrossRefPubMedGoogle Scholar
  28. 28.
    Calin GA, et al. (2005) A microRNA signature associated with prognosis and progression in chronic lymphocytic leukemia. N. Engl. J. Med. 353:1793–801.CrossRefPubMedGoogle Scholar
  29. 29.
    Rossi S, et al. (2010) microRNA fingerprinting of CLL patients with chromosome 17p deletion identify a miR-21 score that stratifies early survival. Blood. 116:945–52.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Ferracin M, et al. (2010) MicroRNAs involvement in fludarabine refractory chronic lymphocytic leukemia. Mol. Cancer. 9:123.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Moussay E, et al. (2010) Determination of genes and microRNAs involved in the resistance to fludarabine in vivo in chronic lymphocytic leukemia. Mol. Cancer. 9:115.CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Frenquelli M, et al. (2010) MicroRNA and proliferation control in chronic lymphocytic leukemia: functional relationship between miR-221/222 cluster and p27. Blood. 115:3949–59.CrossRefPubMedGoogle Scholar
  33. 33.
    Wang M, et al. (2008) miRNA analysis in B-cell chronic lymphocytic leukaemia: proliferation centres characterized by low miR-150 and high BIC/miR-155 expression. J Pathol 215:13–20.CrossRefPubMedGoogle Scholar
  34. 34.
    Hallek M, et al. (2008) Guidelines for the diagnosis and treatment of chronic lymphocytic leukemia: a report from the International Workshop on Chronic Lymphocytic Leukemia updating the National Cancer Institute-Working Group 1996 guidelines. Blood. 111:5446–56.CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Akao Y, et al. (2007) Downregulation of microR-NAs-143 and -145 in B-cell malignancies. Cancer Sci. 98:1914–20.CrossRefPubMedGoogle Scholar
  36. 36.
    Stevenson FK, et al. (2011) B-cell receptor signaling in chronic lymphocytic leukemia. Blood. 118:4313–20.CrossRefPubMedGoogle Scholar
  37. 37.
    Li S, et al. (2009) MicroRNA-101 regulates expression of the v-fos FBJ murine osteosarcoma viral oncogene homolog (FOS) oncogene in human hepatocellular carcinoma. Hepatology 49:1194–202.CrossRefPubMedGoogle Scholar
  38. 38.
    Wang HJ, et al. (2010) MicroRNA-101 is down-regulated in gastric cancer and involved in cell migration and invasion. Eur. J. Cancer. 46:2295–303.CrossRefPubMedGoogle Scholar
  39. 39.
    Morin RD, et al. (2010) Somatic mutations altering EZH2 (Tyr641) in follicular and diffuse large B-cell lymphomas of germinal-center origin. Nat. Genet. 42:181–5.CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Varambally S, et al. (2008) Genomic loss of microRNA-101 leads to overexpression of histone methyltransferase EZH2 in cancer. Science. 322:1695–9.CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Ryan RJ, et al. (2011) EZH2 codon 641 mutations are common in BCL2-rearranged germinal center B cell lymphomas. PLoS One. 6:e28585.CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Cao P, et al. (2010) MicroRNA-101 negatively regulates Ezh2 and its expression is modulated by androgen receptor and HIF-1alpha/HIF-1beta. Mol. Cancer. 9:108.CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Muzio M, et al. (2008) Constitutive activation of distinct BCR-signaling pathways in a subset of CLL patients: a molecular signature of anergy. Blood. 112:188–95.CrossRefPubMedGoogle Scholar
  44. 44.
    Bomben R, et al. (2012) The miR-17∼92 family regulates the response to Toll-like receptor 9 triggering of CLL cells with unmutated IGHV genes. Leukemia. 26:1584–93.CrossRefPubMedGoogle Scholar
  45. 45.
    Kluiver JL, Chen CZ. (2012) MicroRNAs regulate B-cell receptor signaling-induced apoptosis. Genes Immun. 13:239–44.CrossRefPubMedGoogle Scholar
  46. 46.
    Smonskey MT, et al. (2012) Monoallelic and biallelic deletions of 13q14.3 in chronic lymphocytic leukemia: FISH vs miRNA RT-qPCR detection. Am. J. Clin. Pathol. 137:641–6.CrossRefPubMedGoogle Scholar
  47. 47.
    Kostareli E, Gounari M, Agathangelidis A, Stamatopoulos K. (2012) Immunoglobulin gene repertoire in chronic lymphocytic leukemia: insight into antigen selection and microenvironmental interactions. Mediterr. J. Hematol. Infect. Dis. 4:e2012052.CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Simon JA, Lange CA. (2008) Roles of the EZH2 histone methyltransferase in cancer epigenetics. Mutat. Res. 647:21–9.CrossRefPubMedGoogle Scholar
  49. 49.
    Tsang DP, Cheng AS. (2011) Epigenetic regulation of signaling pathways in cancer: role of the histone methyltransferase EZH2. J. Gastroenterol. Hepatol. 26:19–27.CrossRefPubMedGoogle Scholar
  50. 50.
    Guo H, Ingolia NT, Weissman JS, Bartel DP. (2010) Mammalian microRNAs predominantly act to decrease target mRNA levels. Nature. 466:835–40.CrossRefPubMedPubMedCentralGoogle Scholar
  51. 51.
    Baek D, et al. (2008) The impact of microRNAs on protein output. Nature. 455:64–71.CrossRefPubMedPubMedCentralGoogle Scholar
  52. 52.
    Ntoufa S. (2012) The miR-17∼92 cluster is an immunomodulator in CLL regulating distinct functional responses to Toll-like receptors in subsets with stereotyped antigen receptors. Blood (ASH Annual Meeting Abstracts). 120:3862.CrossRefGoogle Scholar

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Authors and Affiliations

  • Nikos Papakonstantinou
    • 1
    • 2
    • 3
  • Stavroula Ntoufa
    • 2
    • 3
  • Elisavet Chartomatsidou
    • 2
    • 3
  • Giorgio Papadopoulos
    • 4
  • Artemis Hatzigeorgiou
    • 4
  • Achiles Anagnostopoulos
    • 2
  • Katerina Chlichlia
    • 1
  • Paolo Ghia
    • 5
  • Marta Muzio
    • 6
  • Chrysoula Belessi
    • 7
  • Kostas Stamatopoulos
    • 2
    • 3
  1. 1.Laboratory of Molecular Immunobiology, Department of Molecular Biology and GeneticsDemocritus University of ThraceAlexandroupolisGreece
  2. 2.Hematology Department and HCT UnitG. Papanicolaou HospitalThessalonikiGreece
  3. 3.Institute of Applied BiosciencesCenter for Research and Technology HellasThessalonikiGreece
  4. 4.Institute of Molecular OncologyBiomedical Sciences Research Center “Alexander Fleming”VariGreece
  5. 5.Laboratory of B cell Neoplasia and Unit of Lymphoid Malignancies, Department of Onco-Hematology and Division of Molecular Oncology, Istituto Scientifico San RaffaeleUniversità Vita-Salute San RaffaeleMilanItaly
  6. 6.Cell Activation and Signaling Unit, Division of Molecular OncologyIstituto Scientifico San RaffaeleMilanItaly
  7. 7.Hematology DepartmentNikea General HospitalAthensGreece

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