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Investigational New Drugs

, Volume 36, Issue 5, pp 955–960 | Cite as

GLP overexpression is associated with poor prognosis in Chronic Lymphocytic Leukemia and its inhibition induces leukemic cell death

  • Juliana Carvalho Alves-Silva
  • Juliana Lott de Carvalho
  • Doralina Amaral Rabello
  • Teresa Raquel Tavares Serejo
  • Eduardo Magalhaes Rego
  • Francisco Assis Rocha Neves
  • Antonio Roberto Lucena-Araujo
  • Fábio Pittella-Silva
  • Felipe Saldanha-Araujo
SHORT REPORT
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Summary

Background Heterodimeric methyltransferases GLP (EHMT1/KMT1D) and G9a (EHMT2/KMT1C) are two closely related enzymes that promote the monomethylation and dimethylation of histone H3 lysine 9. Dysregulation of their activity has been implicated in several types of human cancer. Patients and methods Here, in order to investigate whether GLP/G9a exerts any impact on Chronic Lymphocytic Leukemia (CLL), GLP/G9a expression levels were assessed in a cohort of 50 patients and the effects of their inhibition were verified for the viability of CLL cells. Also, qRT-PCR was used to investigate the transcriptional levels of GLP/G9a in CLL patients. In addition, patient samples were classified according to ZAP-70 protein expression by flow cytometry and according to karyotype integrity by cytogenetics analysis. Finally, a selective small molecule inhibitor for GLP/G9a was used to ascertain whether these methyltransferases influenced the viability of MEC-1 CLL cell lineage. Results mRNA analysis revealed that CLL samples had higher levels of GLP, but not G9a, when compared to non-leukemic controls. Interestingly, patients with unfavorable cytogenetics showed higher expression levels of GLP compared to patients with favorable karyotypes. More importantly, GLP/G9a inhibition markedly induced cell death in CLL cells. Conclusion Taken together, these results indicate that GLP is associated with a worse prognosis in CLL, and that the inhibition of GLP/G9a influences CLL cell viability. Altogether, the present data demonstrate that these methyltransferases can be potential markers for disease progression, as well as a promising epigenetic target for CLL treatment and the prevention of disease evolution.

Keywords

Chronic lymphocytic leukemia GLP G9a UNC0646 Prognostic marker, cytogenetic abnormalities 

Notes

Acknowledgments

This study was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) and Fundação de Amparo à Pesquisa do Distrito Federal (FAPDF).

Compliance with ethical standards

Conflicts of Interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

References

  1. 1.
    Rai KR, Jain P (2016) Chronic lymphocytic leukemia (CLL)-Then and now. Am J Hematol 91:330–340CrossRefPubMedGoogle Scholar
  2. 2.
    Zenz T, Mertens D, Küppers R et al (2009) From pathogenesis to treatment of chronic lymphocytic leukaemia. Nat Rev Cancer 10:37–50CrossRefPubMedGoogle Scholar
  3. 3.
    Rai KR, Sawitsky A, Cronkite EP et al (1975) Clinical staging of chronic lymphocytic leukemia. Blood 46:219–234PubMedGoogle Scholar
  4. 4.
    Binet JL, Auquier A, Dighiero G et al (1981) A new prognostic classification of chronic lymphocytic leukemia derived from a multivariate survival analysis. Cancer 48:198–206CrossRefPubMedGoogle Scholar
  5. 5.
    Scarfò L, Ferreri AJM, Ghia P (2016) Chronic lymphocytic leukaemia. Crit Rev Oncol Hematol 104:169–182CrossRefPubMedGoogle Scholar
  6. 6.
    Roos-Weil D, Nguyen-Khac F, Bernard OA (2016) Chronic lymphocytic leukemia: Time to go past genomics? Am J Hematol 91:518–528CrossRefPubMedGoogle Scholar
  7. 7.
    Kurdistani SK (2007) Histone modifications as markers of cancer prognosis: a cellular view. Br J Cancer 97:1–5CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Mozzetta C, Pontis J, Ait-Si-Ali S (2015) Functional Crosstalk Between Lysine Methyltransferases on Histone Substrates: The Case of G9A/GLP and Polycomb Repressive Complex 2. Antioxid Redox Signal 22:1365–1381CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Matutes E, Owusu-Ankomah K, Morilla R et al (1994) The immunological profile of B-cell disorders and proposal of a scoring system for the diagnosis of CLL. Leukemia 8:1640–1645PubMedGoogle Scholar
  10. 10.
    Crespo M, Bosch F, Villamor N et al (2003) ZAP-70 Expression as a Surrogate for Immunoglobulin-Variable-Region Mutations in Chronic Lymphocytic Leukemia. N Engl J Med 348:1764–1775CrossRefPubMedGoogle Scholar
  11. 11.
    Döhner H, Stilgenbauer S, Benner A et al (2000) Genomic Aberrations and Survival in Chronic Lymphocytic Leukemia. N Engl J Med 343:1910–1916CrossRefPubMedGoogle Scholar
  12. 12.
    Nabhan C, Raca G, Wang YL (2015) Predicting Prognosis in Chronic Lymphocytic Leukemia in the Contemporary Era. JAMA Oncol 1:965–974CrossRefPubMedGoogle Scholar
  13. 13.
    Mayr C, Speicher MR, Kofler DM et al (2006) Chromosomal translocations are associated with poor prognosis in chronic lymphocytic leukemia. Blood 107:742–751CrossRefPubMedGoogle Scholar
  14. 14.
    Guièze R, Wu CJ (2015) Genomic and epigenomic heterogeneity in chronic lymphocytic leukemia. Blood 126:445–453CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Shinkai Y, Tachibana M (2011) H3K9 methyltransferase G9a and the related molecule GLP. Genes Dev 25:781–788CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Loh SW, Ng WL, Yeo KS et al (2014) Inhibition of euchromatic histone methyltransferase 1 and 2 sensitizes chronic myeloid leukemia cells to interferon treatment. PLoS One 9:e103915CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Pappano WN, Guo J, He Y et al (2015) The Histone Methyltransferase Inhibitor A-366 Uncovers a Role for G9a/GLP in the Epigenetics of Leukemia. PLoS One 10:e0131716CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Huang J, Dorsey J, Chuikov S et al (2010) G9a and Glp methylate lysine 373 in the tumor suppressor p53. J Biol Chem 285:9636–9641CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Orchard JA, Ibbotson RE, Davis Z et al (2004) ZAP-70 expression and prognosis in chronic lymphocytic leukaemia. Lancet 363:105–111CrossRefPubMedGoogle Scholar
  20. 20.
    Zenz T, Eichhorst B, Busch R et al (2010) TP53Mutation and Survival in Chronic Lymphocytic Leukemia. J Clin Oncol 28:4473–4479CrossRefPubMedGoogle Scholar
  21. 21.
    Koníková E, Kusenda J (2003) Altered expression of p53 and MDM2 proteins in hematological malignancies. Neoplasma 50:31–40PubMedGoogle Scholar
  22. 22.
    Tomasini R, Mak TW, Melino G (2008) The impact of p53 and p73 on aneuploidy and cancer. Trends Cell Biol 18:244–252CrossRefPubMedGoogle Scholar
  23. 23.
    Lazarian G, Tausch E, Eclache V et al (2016) TP53 mutations are early events in chronic lymphocytic leukemia disease progression and precede evolution to complex karyotypes. Int J Cancer 139:1759–1763CrossRefPubMedGoogle Scholar
  24. 24.
    Dicker F, Herholz H, Schnittger S et al (2009) The detection of TP53 mutations in chronic lymphocytic leukemia independently predicts rapid disease progression and is highly correlated with a complex aberrant karyotype. Leukemia 23:117–124CrossRefPubMedGoogle Scholar
  25. 25.
    Lin X, Huang Y, Zou Y et al (2016) Depletion of G9a gene induces cell apoptosis in human gastric carcinoma. Oncol Rep 35:3041–3049CrossRefPubMedGoogle Scholar
  26. 26.
    Ren A, Qiu Y, Cui H, Fu G (2015) Inhibition of H3K9 methyltransferase G9a induces autophagy and apoptosis in oral squamous cell carcinoma. Biochem Biophys Res Commun 459:10–17CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Li F, Zeng J, Gao Y et al (2015) G9a Inhibition Induces Autophagic Cell Death via AMPK/mTOR Pathway in Bladder Transitional Cell Carcinoma. PLoS One 10:e0138390CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Lai Y-S, Chen J-Y, Tsai H-J et al (2015) The SUV39H1 inhibitor chaetocin induces differentiation and shows synergistic cytotoxicity with other epigenetic drugs in acute myeloid leukemia cells. Blood Cancer J 5:e313CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Juliana Carvalho Alves-Silva
    • 1
    • 2
  • Juliana Lott de Carvalho
    • 3
  • Doralina Amaral Rabello
    • 1
  • Teresa Raquel Tavares Serejo
    • 2
  • Eduardo Magalhaes Rego
    • 4
  • Francisco Assis Rocha Neves
    • 2
  • Antonio Roberto Lucena-Araujo
    • 5
  • Fábio Pittella-Silva
    • 1
  • Felipe Saldanha-Araujo
    • 2
  1. 1.Laboratório de Patologia Molecular do CâncerUniversidade de BrasíliaBrasíliaBrazil
  2. 2.Laboratório de Farmacologia MolecularUniversidade de BrasíliaBrasíliaBrazil
  3. 3.Laboratório de BiotecnologiaUniversidade Católica de BrasíliaBrasíliaBrazil
  4. 4.Laboratório de HematologiaUniversidade de São PauloRibeirão PretoBrazil
  5. 5.Laboratório de HematologiaUniversidade Federal de PernambucoRecifeBrazil

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