Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Molecular Targets for Therapy

Efficacy of cyclin-dependent-kinase 9 inhibitors in a murine model of mixed-lineage leukemia

Leukemia volume 28, pages 1427–1435 (2014)Cite this article

Subjects

Abstract

Mixed-lineage leukemia fusion proteins activate their target genes predominantly by stimulating transcriptional elongation. A core component necessary for this activity is cyclin-dependent kinase 9. Here we explored the effectiveness of small molecules targeting this enzyme as potential therapeutics. A screen of seven compounds with anti-CDK9 activity applied to a panel of leukemia cell lines identified flavopiridol and the experimental inhibitor PC585 as superior in efficacy with inhibitory concentrations in the submicromolar range. Both substances induced rapid dephosphorylation of the RNA polymerase II C-terminal domain, accompanied by downregulation of CDK9-dependent transcripts for MYC and HOXA9. Global gene expression analysis indicated the induction of a general stress response program, culminating in widespread apoptosis. Importantly, colony-forming activity in leukemia lines and primary patient samples could be completely inhibited under conditions that did not affect native precursors from bone marrow. In vivo application in a mouse transplant model significantly delayed disease with PC585 showing also oral activity. These results suggest CDK9 inhibition as novel treatment option for mixed-lineage leukemia.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  1. Krauter J, Wagner K, Schafer I, Marschalek R, Meyer C, Heil G et al. Prognostic factors in adult patients up to 60 years old with acute myeloid leukemia and translocations of chromosome band 11q23: individual patient data-based meta-analysis of the German Acute Myeloid Leukemia Intergroup. J Clin Oncol 2009; 27: 3000–3006.

    Article  PubMed  Google Scholar 

  2. Pui CH, Carroll WL, Meshinchi S, Arceci RJ . Biology, risk stratification, and therapy of pediatric acute leukemias: an update. J Clin Oncol 2011; 29: 551–565.

    Article  PubMed  Google Scholar 

  3. de Boer J, Walf-Vorderwulbecke V, Williams O . In focus: MLL-rearranged leukemia. Leukemia 2013; 27: 1224–1228.

    Article  CAS  PubMed  Google Scholar 

  4. Slany RK . The molecular biology of mixed lineage leukemia. Haematologica 2009; 94: 984–993.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Lin C, Smith ER, Takahashi H, Lai KC, Martin-Brown S, Florens L et al. AFF4, a component of the ELL/P-TEFb elongation complex and a shared subunit of MLL chimeras, can link transcription elongation to leukemia. Mol Cell 2010; 37: 429–437.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Mueller D, Bach C, Zeisig D, Garcia-Cuellar MP, Monroe S, Sreekumar A et al. A role for the MLL fusion partner ENL in transcriptional elongation and chromatin modification. Blood 2007; 110: 4445–4454.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Mueller D, Garcia-Cuellar MP, Bach C, Buhl S, Maethner E, Slany RK . Misguided transcriptional elongation causes mixed lineage leukemia. PLoS Biol 2009; 7: e1000249.

    Article  PubMed  PubMed Central  Google Scholar 

  8. Yokoyama A, Lin M, Naresh A, Kitabayashi I, Cleary ML . A higher-order complex containing AF4 and ENL family proteins with P-TEFb facilitates oncogenic and physiologic MLL-dependent transcription. Cancer Cell 2010; 17: 198–212.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Zeisig BB, Milne T, Garcia-Cuellar MP, Schreiner S, Martin ME, Fuchs U et al. Hoxa9 and Meis1 are key targets for MLL-ENL-mediated cellular immortalization. Mol Cell Biol 2004; 24: 617–628.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Adelman K, Lis JT . Promoter-proximal pausing of RNA polymerase II: emerging roles in metazoans. Nat Rev Genet 2012; 13: 720–731.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Peterlin BM, Price DH . Controlling the elongation phase of transcription with P-TEFb. Mol Cell 2006; 23: 297–305.

    Article  CAS  PubMed  Google Scholar 

  12. Mohan M, Herz HM, Takahashi YH, Lin C, Lai KC, Zhang Y et al. Linking H3K79 trimethylation to Wnt signaling through a novel Dot1-containing complex (DotCom). Genes Dev 2010; 24: 574–589.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Maethner E, Garcia-Cuellar MP, Breitinger C, Takacova S, Divoky V, Hess JL et al. MLL-ENL inhibits polycomb repressive complex 1 to achieve efficient transformation of hematopoietic cells. Cell Rep 2013; 3: 1553–1566.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Daigle SR, Olhava EJ, Therkelsen CA, Basavapathruni A, Jin L, Boriack-Sjodin PA et al. Potent inhibition of DOT1L as treatment of MLL-fusion leukemia. Blood 2013; 122: 1017–1025.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Daigle SR, Olhava EJ, Therkelsen CA, Majer CR, Sneeringer CJ, Song J et al. Selective killing of mixed lineage leukemia cells by a potent small-molecule DOT1L inhibitor. Cancer Cell 2011; 20: 53–65.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Dawson MA, Prinjha RK, Dittmann A, Giotopoulos G, Bantscheff M, Chan WI et al. Inhibition of BET recruitment to chromatin as an effective treatment for MLL-fusion leukaemia. Nature 2011; 478: 529–533.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Delmore JE, Issa GC, Lemieux ME, Rahl PB, Shi J, Jacobs HM et al. BET bromodomain inhibition as a therapeutic strategy to target c-Myc. Cell 2011; 146: 904–917.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Mertz JA, Conery AR, Bryant BM, Sandy P, Balasubramanian S, Mele DA et al. Targeting MYC dependence in cancer by inhibiting BET bromodomains. Proc Natl Acad Sci USA 2011; 108: 16669–16674.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Zuber J, Shi J, Wang E, Rappaport AR, Herrmann H, Sison EA et al. RNAi screen identifies Brd4 as a therapeutic target in acute myeloid leukaemia. Nature 2011; 478: 524–528.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Lavau C, Szilvassy SJ, Slany R, Cleary ML . Immortalization and leukemic transformation of a myelomonocytic precursor by retrovirally transduced HRX-ENL. EMBO J 1997; 16: 4226–4237.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Takacova S, Slany R, Bartkova J, Stranecky V, Dolezel P, Luzna P et al. DNA damage response and inflammatory signaling limit the MLL-ENL-induced leukemogenesis in vivo. Cancer Cell 2012; 21: 517–531.

    Article  CAS  PubMed  Google Scholar 

  22. Subramanian A, Tamayo P, Mootha VK, Mukherjee S, Ebert BL, Gillette MA et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci USA 2005; 102: 15545–15550.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Bartholomeeusen K, Xiang Y, Fujinaga K, Peterlin BM . Bromodomain and extra-terminal (BET) bromodomain inhibition activate transcription via transient release of positive transcription elongation factor b (P-TEFb) from 7SK small nuclear ribonucleoprotein. J Biol Chem 2012; 287: 36609–36616.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Bernt KM, Zhu N, Sinha AU, Vempati S, Faber J, Krivtsov AV et al. MLL-rearranged leukemia is dependent on aberrant H3K79 methylation by DOT1L. Cancer Cell 2011; 20: 66–78.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Guenther MG, Lawton LN, Rozovskaia T, Frampton GM, Levine SS, Volkert TL et al. Aberrant chromatin at genes encoding stem cell regulators in human mixed-lineage leukemia. Genes Dev 2008; 22: 3403–3408.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Wilkinson AC, Ballabio E, Geng H, North P, Tapia M, Kerry J et al. RUNX1 is a key target in t(4;11) leukemias that contributes to gene activation through an AF4-MLL complex interaction. Cell Rep 2013; 3: 116–127.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Loven J, Hoke HA, Lin CY, Lau A, Orlando DA, Vakoc CR et al. Selective inhibition of tumor oncogenes by disruption of super-enhancers. Cell 2013; 153: 320–334.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. He N, Pezda AC, Zhou Q . Modulation of a P-TEFb functional equilibrium for the global control of cell growth and differentiation. Mol Cell Biol 2006; 26: 7068–7076.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Napolitano G, Amente S, Castiglia V, Gargano B, Ruda V, Darzacq X et al. Caffeine prevents transcription inhibition and P-TEFb/7SK dissociation following UV-induced DNA damage. PLoS One 2010; 5: e11245.

    Article  PubMed  PubMed Central  Google Scholar 

  30. Ramaswamy B, Phelps MA, Baiocchi R, Bekaii-Saab T, Ni W, Lai JP et al. A dose-finding, pharmacokinetic and pharmacodynamic study of a novel schedule of flavopiridol in patients with advanced solid tumors. Investig New Drugs 2012; 30: 629–638.

    Article  CAS  Google Scholar 

  31. Barr PM . Flavopiridol: Judged by the company one keeps. Leuk Res 2013; 37: 1187–1188.

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Renate Zimmermann for technical assistance and Ingenium Pharmaceuticals for the generous provision with CDK9 inhibitors. This work was supported by research funding from Sander Stiftung, Grant 2010.069.1 to RKS.

Author information

Authors and Affiliations

  1. Department of Genetics, University Erlangen, Erlangen, Germany

    M-P Garcia-Cuellar, E Füller, E Mäthner, C Breitinger, K Hetzner & R K Slany

  2. Ingenium Pharmaceuticals AG, Munich, Germany

    L Zeitlmann

  3. Clinic of Paediatric Oncology, Haematology and Clinical Immunology, Centre for Child and Adolescent Health, Medical Faculty, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany

    A Borkhardt

Authors
  1. M-P Garcia-Cuellar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E Füller
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. E Mäthner
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. C Breitinger
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. K Hetzner
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. L Zeitlmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. A Borkhardt
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. R K Slany
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to R K Slany.

Ethics declarations

Competing interests

L Zeitlmann is an employee of Ingenium Pharmaceuticals. The other authors declare no conflict of interest.

Additional information

Supplementary Information accompanies this paper on the Leukemia website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Garcia-Cuellar, MP., Füller, E., Mäthner, E. et al. Efficacy of cyclin-dependent-kinase 9 inhibitors in a murine model of mixed-lineage leukemia. Leukemia 28, 1427–1435 (2014). https://doi.org/10.1038/leu.2014.40

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/leu.2014.40

Keywords

This article is cited by

Search

Advanced search

Quick links