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

Effective targeting of STAT5-mediated survival in myeloproliferative neoplasms using ABT-737 combined with rapamycin

Abstract

Signal transducer and activator of transcription-5 (STAT5) is a critical transcription factor for normal hematopoiesis and its sustained activation is associated with hematologic malignancy. A persistently active mutant of STAT5 (STAT5aS711F) associates with Grb2-associated binding protein 2 (Gab2) in myeloid leukemias and promotes growth in vitro through AKT activation. Here we have retrovirally transduced wild-type or Gab2−/− mouse bone marrow cells expressing STAT5aS711F and transplanted into irradiated recipient mice to test an in vivo myeloproliferative disease model. To target Gab2-independent AKT/mTOR activation, we treated wild-type mice separately with rapamycin. In either case, mice lacking Gab2 or treated with rapamycin showed attenuated myeloid hyperplasia and modestly improved survival, but the effects were not cytotoxic and were reversible. To improve on this approach, we combined in vitro targeting of STAT5-mediated AKT/mTOR using rapamycin with inhibition of the STAT5 direct target genes bcl-2 and bcl-XL using ABT-737. Striking synergy with both drugs was observed in mouse BaF3 cells expressing STAT5aS711F, TEL-JAK2 or BCR-ABL and in the relatively single agent-resistant human BCR-ABL-positive K562 cell line. Therefore, targeting distinct STAT5-mediated survival signals, for example, bcl-2/bcl-XL and AKT/mTOR may be an effective therapeutic approach for human myeloproliferative neoplasms.

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

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

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

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

References

  1. Iyer J, Reich NC . Constitutive nuclear import of latent and activated STAT5a by its coiled coil domain. FASEB J 2008; 22: 391–400.

    Article  CAS  PubMed  Google Scholar 

  2. Kotecha N, Flores NJ, Irish JM, Simonds EF, Sakai DS, Archambeault S et al. Single-cell profiling identifies aberrant STAT5 activation in myeloid malignancies with specific clinical and biologic correlates. Cancer Cell 2008; 14: 335–343.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Heuser M, Sly LM, Argiropoulos B, Kuchenbauer F, Lai C, Weng A et al. Modelling the functional heterogeneity of leukemia stem cells: role of STAT5 in leukemia stem cell self-renewal. Blood 2009; 114: 3983–3993.

    Article  CAS  PubMed  Google Scholar 

  4. Moellering RE, Cornejo M, Davis TN, Del BC, Aster JC, Blacklow SC et al. Direct inhibition of the NOTCH transcription factor complex. Nature 2009; 462: 182–188.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Gorczynski MJ, Grembecka J, Zhou Y, Kong Y, Roudaia L, Douvas MG et al. Allosteric inhibition of the protein–protein interaction between the leukemia-associated proteins Runx1 and CBFbeta. Chem Biol 2007; 14: 1186–1197.

    Article  CAS  PubMed  Google Scholar 

  6. Schwaller J, Parganas E, Wang D, Cain D, Aster JC, Williams IR et al. Stat5 is essential for the myelo- and lymphoproliferative disease induced by TEL/JAK2. Mol Cell 2000; 6: 693–704.

    Article  CAS  PubMed  Google Scholar 

  7. Kato Y, Iwama A, Tadokoro Y, Shimoda K, Minoguchi M, Akira S et al. Selective activation of STAT5 unveils its role in stem cell self-renewal in normal and leukemic hematopoiesis. J Exp Med 2005; 202: 169–179.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Harir N, Pecquet C, Kerenyi M, Sonneck K, Kovacic B, Nyga R et al. Constitutive activation of Stat5 promotes its cytoplasmic localization and association with PI 3-kinase in myeloid leukemias. Blood 2007; 109: 1678–1686.

    Article  CAS  PubMed  Google Scholar 

  9. Moriggl R, Sexl V, Kenner L, Duntsch C, Stangl K, Gingras S et al. Stat5 tetramer formation is associated with leukemogenesis. Cancer Cell 2005; 7: 87–99.

    Article  CAS  PubMed  Google Scholar 

  10. Li G, Miskimen KL, Wang Z, Xie XY, Brenzovich J, Ryan JJ et al. STAT5 requires the N-domain for suppression of miR15/16, induction of bcl-2, and survival signaling in myeloproliferative disease. Blood 2010; 115: 1416–1424.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Um M, Lodish HF . Antiapoptotic effects of erythropoietin in differentiated neuroblastoma SH-SY5Y cells require activation of both the STAT5 and AKT signaling pathways. J Biol Chem 2006; 281: 5648–5656.

    Article  CAS  PubMed  Google Scholar 

  12. Moon JJ, Rubio ED, Martino A, Krumm A, Nelson BH . A permissive role for phosphatidylinositol 3-kinase in the Stat5-mediated expression of cyclin D2 by the interleukin-2 receptor. J Biol Chem 2004; 279: 5520–5527.

    Article  CAS  PubMed  Google Scholar 

  13. Kirito K, Watanabe T, Sawada K, Endo H, Ozawa K, Komatsu N . Thrombopoietin regulates Bcl-xL gene expression through Stat5 and phosphatidylinositol 3-kinase activation pathways. J Biol Chem 2002; 277: 8329–8337.

    Article  CAS  PubMed  Google Scholar 

  14. Scherr M, Chaturvedi A, Battmer K, Dallmann I, Schultheis B, Ganser A et al. Enhanced sensitivity to inhibition of SHP2, STAT5, and Gab2 expression in chronic myeloid leukemia (CML). Blood 2006; 107: 3279–3287.

    Article  CAS  PubMed  Google Scholar 

  15. Nyga R, Pecquet C, Harir N, Gu H, Dhennin-Duthille I, Regnier A et al. Activated STAT5 proteins induce activation of the PI 3-kinase/Akt and Ras/MAPK pathways via the Gab2 scaffolding adapter. Biochem J 2005; 390: 359–366.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Lockyer HM, Tran E, Nelson BH . STAT5 is essential for Akt/p70S6 kinase activity during IL-2-induced lymphocyte proliferation. J Immunol 2007; 179: 5301–5308.

    Article  CAS  PubMed  Google Scholar 

  17. Gu H, Pratt JC, Burakoff SJ, Neel BG . Cloning of p97/Gab2, the major SHP2-binding protein in hematopoietic cells, reveals a novel pathway for cytokine-induced gene activation. Mol Cell 1998; 2: 729–740.

    Article  CAS  PubMed  Google Scholar 

  18. Nishida K, Yoshida Y, Itoh M, Fukada T, Ohtani T, Shirogane T et al. Gab-family adapter proteins act downstream of cytokine and growth factor receptors and T- and B-cell antigen receptors. Blood 1999; 93: 1809–1816.

    CAS  PubMed  Google Scholar 

  19. Crouin C, Arnaud M, Gesbert F, Camonis J, Bertoglio J . A yeast two-hybrid study of human p97/Gab2 interactions with its SH2 domain-containing binding partners. FEBS Lett 2001; 495: 148–153.

    Article  CAS  PubMed  Google Scholar 

  20. Yu WM, Hawley TS, Hawley RG, Qu CK . Role of the docking protein Gab2 in beta(1)-integrin signaling pathway-mediated hematopoietic cell adhesion and migration. Blood 2002; 99: 2351–2359.

    Article  CAS  PubMed  Google Scholar 

  21. Rosa Santos SC, Dumon S, Mayeux P, Gisselbrecht S, Gouilleux F . Cooperation between STAT5 and phosphatidylinositol 3-kinase in the IL-3-dependent survival of a bone marrow derived cell line. Oncogene 2000; 19: 1164–1172.

    Article  CAS  PubMed  Google Scholar 

  22. Santos SC, Lacronique V, Bouchaert I, Monni R, Bernard O, Gisselbrecht S et al. Constitutively active STAT5 variants induce growth and survival of hematopoietic cells through a PI3-kinase/Akt dependent pathway. Oncogene 2001; 20: 2080–2090.

    Article  CAS  PubMed  Google Scholar 

  23. Li G, Wang Z, Zhang Y, Kang Z, Haviernikova E, Cui Y et al. STAT5 requires the N-domain to maintain hematopoietic stem cell repopulating function and appropriate lymphoid-myeloid lineage output. Exp Hematol 2007; 35: 1684–1694.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Zhang Y, Diaz-Flores E, Li G, Wang Z, Kang Z, Haviernikova E et al. Abnormal hematopoiesis in Gab2 mutant mice. Blood 2007; 110: 116–124.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Yilmaz OH, Valdez R, Theisen BK, Guo W, Ferguson DO, Wu H et al. Pten dependence distinguishes haematopoietic stem cells from leukaemia-initiating cells. Nature 2006; 441: 475–482.

    Article  CAS  PubMed  Google Scholar 

  26. Hoelbl A, Kovacic B, Kerenyi MA, Simma O, Warsch W, Cui Y et al. Clarifying the role of Stat5 in lymphoid development and Abelson induced transformation. Blood 2006; 107: 4898–4906.

    Article  CAS  PubMed  Google Scholar 

  27. Rocnik JL, Okabe R, Yu JC, Giese N, Schenkein DP, Gilliland DG . Roles of tyrosine 589 and 591 in STAT5 activation and transformation mediated by FLT3-ITD. Blood 2006; 108: 1339–1345.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Cain JA, Xiang Z, O’Neal J, Kreisel F, Colson A, Luo H et al. Myeloproliferative disease induced by TEL-PDGFRB displays dynamic range sensitivity to Stat5 gene dosage. Blood 2007; 109: 3906–3914.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Bumm TG, Elsea C, Corbin AS, Loriaux M, Sherbenou D, Wood L et al. Characterization of murine JAK2V617F-positive myeloproliferative disease. Cancer Res 2006; 66: 11156–11165.

    Article  CAS  PubMed  Google Scholar 

  30. Huang J, Manning BD . A complex interplay between Akt, TSC2 and the two mTOR complexes. Biochem Soc Trans 2009; 37: 217–222.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Mohi MG, Boulton C, Gu TL, Sternberg DW, Neuberg D, Griffin JD et al. Combination of rapamycin and protein tyrosine kinase (PTK) inhibitors for the treatment of leukemias caused by oncogenic PTKs. Proc Natl Acad Sci USA 2004; 101: 3130–3135.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Corbacioglu S, Kilic M, Westhoff MA, Reinhardt D, Fulda S, Debatin KM . Newly identified c-KIT receptor tyrosine kinase ITD in childhood AML induces ligand-independent growth and is responsive to a synergistic effect of imatinib and rapamycin. Blood 2006; 108: 3504–3513.

    Article  CAS  PubMed  Google Scholar 

  33. Wei G, Twomey D, Lamb J, Schlis K, Agarwal J, Stam RW et al. Gene expression-based chemical genomics identifies rapamycin as a modulator of MCL1 and glucocorticoid resistance. Cancer Cell 2006; 10: 331–342.

    Article  CAS  PubMed  Google Scholar 

  34. Kim KW, Moretti L, Mitchell LR, Jung DK, Lu B . Combined Bcl-2/mammalian target of rapamycin inhibition leads to enhanced radiosensitization via induction of apoptosis and autophagy in non-small cell lung tumor xenograft model. Clin Cancer Res 2009; 15: 6096–6105.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Ackler S, Xiao Y, Mitten MJ, Foster K, Oleksijew A, Refici M et al. ABT-263 and rapamycin act cooperatively to kill lymphoma cells in vitro and in vivo. Mol Cancer Ther 2008; 7: 3265–3274.

    Article  CAS  PubMed  Google Scholar 

  36. Tse C, Shoemaker AR, Adickes J, Anderson MG, Chen J, Jin S et al. ABT-263: a potent and orally bioavailable Bcl-2 family inhibitor. Cancer Res 2008; 68: 3421–3428.

    Article  CAS  PubMed  Google Scholar 

  37. Yoshimoto G, Miyamoto T, Jabbarzadeh-Tabrizi S, Iino T, Rocnik JL, Kikushige Y et al. FLT3-ITD up-regulates MCL-1 to promote survival of stem cells in acute myeloid leukemia via FLT3-ITD-specific STAT5 activation. Blood 2009; 114: 5034–5043.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Tahir SK, Wass J, Joseph MK, Devanarayan V, Hessler P, Zhang H et al. Identification of expression signatures predictive of sensitivity to the Bcl-2 family member inhibitor ABT-263 in small cell lung carcinoma and leukemia/lymphoma cell lines. Mol Cancer Ther 2010; 9: 545–557.

    Article  CAS  PubMed  Google Scholar 

  39. Janes MR, Limon JJ, So L, Chen J, Lim RJ, Chavez MA et al. Effective and selective targeting of leukemia cells using a TORC1/2 kinase inhibitor. Nat Med 2010; 16: 205–213.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank Henry Koon for critical review of the article and Fang Xu for biostatistical support. We also thank Jalpa Patel and Emma Arriola at Abbott Laboratories for supplying ABT-737. This work was supported by NIH R01DK059380 (KDB), The Center for Stem Cell and Regenerative Medicine, SFB-F28 (RM) and the Flow Cytometry, Radiation Resources, and Histology Core Facilities of the Case Comprehensive Cancer Center (P30CA43703).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to K D Bunting.

Ethics declarations

Competing interests

The material is original research, has not been previously published and has not been submitted for publication elsewhere while under consideration. The authors also have no conflicts of interest to declare regarding the studies performed.

Additional information

Supplementary Information accompanies the paper on the Leukemia website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, G., Miskimen, K., Wang, Z. et al. Effective targeting of STAT5-mediated survival in myeloproliferative neoplasms using ABT-737 combined with rapamycin. Leukemia 24, 1397–1405 (2010). https://doi.org/10.1038/leu.2010.131

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/leu.2010.131

Keywords

This article is cited by

Search

Quick links