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

Targeting the KIT activating switch control pocket: a novel mechanism to inhibit neoplastic mast cell proliferation and mast cell activation

Leukemia volume 27, pages 278–285 (2013)Cite this article

Subjects

Abstract

Activating mutations in the receptor tyrosine kinase KIT, most notably KIT D816V, are commonly observed in patients with systemic mastocytosis. Thus, inhibition of KIT has been a major focus for treatment of this disorder. Here we investigated a novel approach to such inhibition. Utilizing rational drug design, we targeted the switch pocket (SP) of KIT, which regulates its catalytic conformation. Two SP inhibitors thus identified, DP-2976 and DP-4851, were examined for effects on neoplastic mast cell proliferation and mast cell activation. Autophosphorylation of both wild-type and, where also examined, KIT D816V activation was blocked by these compounds in transfected 293T cells, HMC 1.1 and 1.2 human mast cell lines, and in CD34+-derived human mast cells activated by stem cell factor (SCF). Both inhibitors induced apoptosis in the neoplastic mast cell lines and reduced survival of primary bone marrow mast cells from patients with mastocytosis. Moreover, the SP inhibitors more selectively blocked SCF potentiation of FcɛRI-mediated degranulation. Overall, SP inhibitors represent an innovative mechanism of KIT inhibition whose dual suppression of KIT D816V neoplastic mast cell proliferation and SCF-enhanced mast cell activation may provide significant therapeutic benefits.

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
Figure 7

Similar content being viewed by others

References

  1. Escribano L, Ocqueteau M, Almeida J, Orfao A, San Miguel JF . Expression of the c-kit (CD117) molecule in normal and malignant hematopoiesis. Leuk Lymphoma 1998; 30: 459–466.

    Article  CAS  PubMed  Google Scholar 

  2. Jensen BM, Akin C, Gilfillan AM . Pharmacological targeting of the KIT growth factor receptor: a therapeutic consideration for mast cell disorders. Br J Pharmacol 2008; 154: 1572–1582.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Hundley TR, Gilfillan AM, Tkaczyk C, Andrade MV, Metcalfe DD, Beaven MA . Kit and FcɛR1 mediate unique and convergent signals for release of inflammatory mediators from human mast cells. Blood 2004; 104: 2410–2417.

    Article  CAS  PubMed  Google Scholar 

  4. Nagata H, Worobec AS, Oh CK, Chowdhury BA, Tannenbaum S, Suzuki Y et al. Identification of a point mutation in the catalytic domain of the protooncogene c-kit in peripheral blood mononuclear cells of patients who have mastocytosis with an associated hematologic disorder. Proc Natl Acad Sci USA 1995; 92: 10560–10564.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Horny H-P, Metcalfe DD, Bennett JM, Bain BJ, Akin C, Escribano L, Valent. P . Mastocytosis. In Swerdlow SH, Campo E, Harris NL, Jaffee ES, Pileri SA, Stein H, Thiele J, Varderman JW, (eds). WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues 4th edn. International Agency for Research on Cancer: Lyon, France, 2008, 54–63pp.

    Google Scholar 

  6. Verstovsek S, Tefferi A, Cortes J, O′Brien S, Garcia-Manero G, Pardanani A et al. Phase II study of dasatinib in Philadelphia chromosome-negative acute and chronic myeloid diseases, including systemic mastocytosis. Clin Cancer Res 2008; 14: 3906–3915.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Vega-Ruiz A, Cortes JE, Sever M, Manshouri T, Quintas-Cardama A, Luthra R et al. Phase II study of imatinib mesylate as therapy for patients with systemic mastocytosis. Leuk Res 2009; 33: 1481–1484.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Andreas H, Ottmann OG, Lauber S, Hughes T, Verhoef G, Schwarer AP et al. A phase II study of Nilotinib, a novel inhibitor of c-Kit, PDGFR, and Bcr-Abl, administered to patients with systemic mastocytosis. Blood (ASH Annual Meeting Abstracts) 2006; 108: abstract 2703.

  9. Ustun C, DeRemer DL, Akin C . Tyrosine kinase inhibitors in the treatment of systemic mastocytosis. Leuk Res 2011; 35: 1143–1152.

    Article  CAS  PubMed  Google Scholar 

  10. Gajiwala KS, Wu JC, Christensen J, Deshmukh GD, Diehl W, DiNitto JP et al. KIT kinase mutants show unique mechanisms of drug resistance to imatinib and sunitinib in gastrointestinal stromal tumor patients. Proc Natl Acad Sci USA 2009; 106: 1542–1547.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Mol CD, Dougan DR, Schneider TR, Skene RJ, Kraus ML, Scheibe DN et al. Structural basis for the autoinhibition and STI-571 inhibition of c-Kit tyrosine kinase. J Biol Chem 2004; 279: 31655–31663.

    Article  CAS  PubMed  Google Scholar 

  12. Fletcher JA, Rubin BP . KIT mutations in GIST. Curr Opin Genet Dev 2007; 17: 3–7.

    Article  CAS  PubMed  Google Scholar 

  13. Schindler T, Bornmann W, Pellicena P, Miller WT, Clarkson B, Kuriyan J . Structural mechanism for STI-571 inhibition of abelson tyrosine kinase. Science 2000; 289: 1938–1942.

    CAS  PubMed  Google Scholar 

  14. Butterfield JH, Weiler D, Dewald G, Gleich GJ . Establishment of an immature mast cell line from a patient with mast cell leukemia. Leuk Res 1988; 12: 345–355.

    Article  CAS  PubMed  Google Scholar 

  15. Kirshenbaum AS, Goff JP, Semere T, Foster B, Scott LM, Metcalfe DD . Demonstration that human mast cells arise from a progenitor cell population that is CD34(+), c-kit(+), and expresses aminopeptidase N (CD13). Blood 1999; 94: 2333–2342.

    CAS  PubMed  Google Scholar 

  16. Caruana G, Cambareri AC, Ashman LK . Isoforms of c-KIT differ in activation of signalling pathways and transformation of NIH3T3 fibroblasts. Oncogene 1999; 18: 5573–5581.

    Article  CAS  PubMed  Google Scholar 

  17. Iwaki S, Spicka J, Tkaczyk C, Jensen BM, Furumoto Y, Charles N et al. Kit- and Fc epsilonRI-induced differential phosphorylation of the transmembrane adaptor molecule NTAL/LAB/LAT2 allows flexibility in its scaffolding function in mast cells. Cell Signal 2008; 20: 195–205.

    Article  CAS  PubMed  Google Scholar 

  18. Tkaczyk C, Metcalfe DD, Gilfillan AM . Determination of protein phosphorylation in Fc epsilon RI-activated human mast cells by immunoblot analysis requires protein extraction under denaturing conditions. J Immunol Methods 2002; 268: 239–243.

    Article  CAS  PubMed  Google Scholar 

  19. Jensen BM, Beaven MA, Iwaki S, Metcalfe DD, Gilfillan AM . Concurrent inhibition of kit- and FcepsilonRI-mediated signaling: coordinated suppression of mast cell activation. J Pharmacol Exp Ther 2008; 324: 128–138.

    Article  CAS  PubMed  Google Scholar 

  20. Woolhiser MR, Okayama Y, Gilfillan AM, Metcalfe DD . IgG-dependent activation of human mast cells following up-regulation of FcgammaRI by IFN-gamma. Eur J Immunol 2001; 31: 3298–3307.

    Article  CAS  PubMed  Google Scholar 

  21. Shah NP, Lee FY, Luo R, Jiang Y, Donker M, Akin C . Dasatinib (BMS-354825) inhibits KITD816V, an imatinib-resistant activating mutation that triggers neoplastic growth in most patients with systemic mastocytosis. Blood 2006; 108: 286–291.

    Article  CAS  PubMed  Google Scholar 

  22. Akin C, Brockow K, D′Ambrosio C, Kirshenbaum AS, Ma Y, Longley BJ et al. Effects of tyrosine kinase inhibitor STI571 on human mast cells bearing wild-type or mutated c-kit. Exp Hematol 2003; 31: 686–692.

    Article  CAS  PubMed  Google Scholar 

  23. Gleixner KV, Mayerhofer M, Aichberger KJ, Derdak S, Sonneck K, Bohm A et al. PKC412 inhibits in vitro growth of neoplastic human mast cells expressing the D816V-mutated variant of KIT: comparison with AMN107, imatinib, and cladribine (2CdA) and evaluation of cooperative drug effects. Blood 2006; 107: 752–759.

    Article  CAS  PubMed  Google Scholar 

  24. Krauth MT, Mirkina I, Herrmann H, Baumgartner C, Kneidinger M, Valent P . Midostaurin (PKC412) inhibits immunoglobulin E-dependent activation and mediator release in human blood basophils and mast cells. Clin Exp Allergy 2009; 39: 1711–1720.

    Article  CAS  PubMed  Google Scholar 

  25. Eide CA, Adrian LT, Tyner JW, Mac Partlin M, Anderson DJ, Wise SC et al. The ABL switch control inhibitor DCC-2036 is active against the chronic myeloid leukemia mutant BCR-ABLT315I and exhibits a narrow resistance profile. Cancer Res 2011; 71: 3189–3195.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Chan WW, Wise SC, Kaufman MD, Ahn YM, Ensinger CL, Haack T et al. Conformational control inhibition of the BCR-ABL1 tyrosine kinase, including the gatekeeper T315I mutant, by the switch-control inhibitor DCC-2036. Cancer Cell 2011; 19: 556–568.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Azam M, Seeliger MA, Gray NS, Kuriyan J, Daley GQ . Activation of tyrosine kinases by mutation of the gatekeeper threonine. Nat Struct Mol Biol 2008; 15: 1109–1118.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Gotlib J, Berube C, Growney JD, Chen CC, George TI, Williams C et al. Activity of the tyrosine kinase inhibitor PKC412 in a patient with mast cell leukemia with the D816V KIT mutation. Blood 2005; 106: 2865–2870.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Gotlib J, DeAngelo DJ, George TI, Corless CL, Linder A, Langford C et al. KIT Inhibitor Midostaurin exhibits a high rate of clinically meaningful and durable responses in advanced systemic mastocytosis: report of a fully accrued phase II trial. Blood (ASH Annual Meeting Abstracts) 2010; 116: abstract 316.

  30. Tefferi A, Levine RL, Lim KH, Abdel-Wahab O, Lasho TL, Patel J et al. Frequent TET2 mutations in systemic mastocytosis: clinical, KITD816V and FIP1L1-PDGFRA correlates. Leukemia 2009; 23: 900–904.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Wilson TM, Maric I, Simakova O, Bai Y, Chan EC, Olivares N et al. Clonal analysis of NRAS activating mutations in KIT-D816V systemic mastocytosis. Haematologica 2011; 96: 459–463.

    Article  CAS  PubMed  Google Scholar 

  32. Sotlar K, Bache A, Stellmacher F, Bultmann B, Valent P, Horny HP . Systemic mastocytosis associated with chronic idiopathic myelofibrosis: a distinct subtype of systemic mastocytosis associated with a [corrected] clonal hematological non-mast [corrected] cell lineage disorder carrying the activating point mutations KITD816V and JAK2V617F. J Mol Diagn 2008; 10: 58–66.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Gleixner KV, Mayerhofer M, Sonneck K, Gruze A, Samorapoompichit P, Baumgartner C et al. Synergistic growth-inhibitory effects of two tyrosine kinase inhibitors, dasatinib and PKC412, on neoplastic mast cells expressing the D816V-mutated oncogenic variant of KIT. Haematologica 2007; 92: 1451–1459.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Dr Cem Akin (Brigham and Women’s Hospital) for technical advice with the ex vivo studies and Sarka Smrzova (LAD) for technical support with HuMC cultures. We thank all the subjects as well as the LAD clinical research staff for their contributions. This study was supported in part by the Division of Intramural Research of the NIAID, NIH. Deciphera Pharmaceuticals supplied the study compounds and performed the in vitro kinase assays.

Author contributions

YB, GB, SNB, ECC, OS, W-PL and TMW performed the research. TMW, AMG, DDM, IM, SCW and DLF designed the research. TMW and DDM recruited and cared for the patients. YB, GB, ECC, OS, IM, SCW, DLF, DDM and TMW analyzed and interpreted the data. YB, GB, AMG and TMW wrote the paper with edits from all authors.

Author information

Author notes

  1. Y Bai and G Bandara: These authors share first authorship.

Authors and Affiliations

  1. Laboratory of Allergic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA

    Y Bai, G Bandara, E Ching Chan, S N Bandara, D D Metcalfe, A M Gilfillan & T M Wilson

  2. Department of Laboratory Medicine, Clinical Center, Bethesda, MD, USA

    I Maric & O Simakova

  3. Deciphera Pharmaceuticals, Lawrence, KS, USA

    W-P Lu, S C Wise & D L Flynn

Authors
  1. Y Bai
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G Bandara
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. E Ching Chan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. I Maric
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. O Simakova
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. S N Bandara
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. W-P Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. S C Wise
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. D L Flynn
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. D D Metcalfe
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. A M Gilfillan
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. T M Wilson
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to T M Wilson.

Ethics declarations

Competing interests

W-PL, SCW and DLF are employees of Deciphera Pharmaceuticals, which is the owner of DP-2976 and DP-4851, the compounds studied and reported in this work. The NIH authors declare no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bai, Y., Bandara, G., Ching Chan, E. et al. Targeting the KIT activating switch control pocket: a novel mechanism to inhibit neoplastic mast cell proliferation and mast cell activation. Leukemia 27, 278–285 (2013). https://doi.org/10.1038/leu.2012.218

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/leu.2012.218

Keywords

This article is cited by

Search

Advanced search

Quick links