Identification of Ponatinib as a potent inhibitor of growth, migration, and activation of neoplastic eosinophils carrying FIP1L1-PDGFRA - PubMed (original) (raw)
Clinical Trial
. 2014 Apr;42(4):282-293.e4.
doi: 10.1016/j.exphem.2013.12.007. Epub 2014 Jan 6.
Els Lierman 2, Barbara Peter 3, Harald Herrmann 3, Verena Suppan 1, Gabriele Stefanzl 1, Oskar Haas 4, Thomas Lion 4, Winfried Pickl 5, Jan Cools 2, Peter Vandenberghe 2, Peter Valent 6
Affiliations
- PMID: 24407160
- PMCID: PMC4338611
- DOI: 10.1016/j.exphem.2013.12.007
Clinical Trial
Identification of Ponatinib as a potent inhibitor of growth, migration, and activation of neoplastic eosinophils carrying FIP1L1-PDGFRA
Irina Sadovnik et al. Exp Hematol. 2014 Apr.
Abstract
In chronic eosinophilic leukemia, the transforming oncoprotein FIP1L1-PDGFRA is a major target of therapy. In most patients, the tyrosine kinase inhibitor (TKI) imatinib induces complete remission. For patients who are intolerant or resistant, novel TKIs have been proposed. We examined the in vitro effects of 14 kinase blockers on growth and function of EOL-1 cells, a FIP1L1-PDGFRA(+) eosinophil cell line. Major growth-inhibitory effects were seen with all PDGFR-blocking agents, with IC50 values in the low nanomolar range: ponatinib, 0.1-0.2 nmol/L; sorafenib, 0.1-0.2 nmol/L; masitinib, 0.2-0.5 nmol/L; nilotinib, 0.2-1.0 nmol/L; dasatinib, 0.5-2.0 nmol/L; sunitinib, 1-2 nmol/L; midostaurin, 5-10 nmol/L. These drugs were also found to block activation of PDGFR-downstream signaling molecules, including Akt, S6, and STAT5 in EOL-1 cells. All effective TKIs produced apoptosis in EOL-1 cells as determined by microscopy, Annexin-V/PI, and caspase-3 staining. In addition, PDGFR-targeting TKIs were found to inhibit cytokine-induced migration of EOL-1 cells. In all bioassays used, ponatinib was found to be the most potent compound in EOL-1 cells. In addition, ponatinib was found to downregulate expression of the activation-linked surface antigen CD63 on EOL-1 cells and to suppress the growth of primary neoplastic eosinophils. We also examined drug effects on Ba/F3 cells expressing two clinically relevant, imatinib-resistant, mutant forms of FIP1L1-PDGFRA, namely T674I and D842V. Strong inhibitory effects on both mutants were seen only with ponatinib. In summary, novel PDGFR-targeting TKIs may be alternative agents for the treatment of patients with imatinib-resistant chronic eosinophilic leukemia. Although several different PDGFR-targeting agents are effective, the most potent drug appears to be ponatinib.
Copyright © 2014 ISEH - Society for Hematology and Stem Cells. Published by Elsevier Inc. All rights reserved.
Figures
Figure 1. Effects of various kinase blockers on proliferation of neoplastic eosinophils
(A) EOL-1 cells were incubated in control medium (Co) or in various concentrations of targeted drugs as indicated at 37°C for 48 hours. After incubation, 3H-thymidine uptake was measured. Results are expressed as percent of control and represent the mean±SD from 3 independent experiments. Asterisk: p<0.05. (B) Primary eosinophils obtained from a patient with FIP1L1-PDGFRA+ CEL (upper panels), one with aggressive systemic mastocytosis with eosinophilia (ASM-eo; middle panels) and one with reactive hypereosinophilia (HER, lower panels), were incubated in control medium (Co) or in various concentrations of targeted drugs as indicated at 37°C for 48 hours. After incubation, 3H-thymidine uptake was measured. Results are expressed as percent of control and represent the mean±SD of triplicates.
Figure 1. Effects of various kinase blockers on proliferation of neoplastic eosinophils
(A) EOL-1 cells were incubated in control medium (Co) or in various concentrations of targeted drugs as indicated at 37°C for 48 hours. After incubation, 3H-thymidine uptake was measured. Results are expressed as percent of control and represent the mean±SD from 3 independent experiments. Asterisk: p<0.05. (B) Primary eosinophils obtained from a patient with FIP1L1-PDGFRA+ CEL (upper panels), one with aggressive systemic mastocytosis with eosinophilia (ASM-eo; middle panels) and one with reactive hypereosinophilia (HER, lower panels), were incubated in control medium (Co) or in various concentrations of targeted drugs as indicated at 37°C for 48 hours. After incubation, 3H-thymidine uptake was measured. Results are expressed as percent of control and represent the mean±SD of triplicates.
Figure 2. PDGFR-targeting drugs induce apoptosis in EOL-1 cells
EOL-1 cells were cultured in the absence (Co) or presence of various concentrations of targeted drugs as indicated at 37°C for 48 hours. Apoptosis was determined by Annexin-V/PI-staining by flow cytometry. Results are expressed as percent of control and represent the mean±S.D. of 3 or 4 independent experiments. Asterisk: p<0.05. In the lower right panel, the DMSO control is also shown.
Figure 3. Effects of various targeted drugs on PDGFR-downstream signaling molecules in EOL-1 cells
(A) EOL-1 cells were incubated in control medium in the absence (Co) or presence of ponatinib, sorafenib, masitinib, sunitinib or erlotinib (each 0.01, 0.1 or 1 μM) at 37°C for 4 hours. After incubation, cells were analyzed for expression of pAKT, pS6, and pSTAT5, by flow cytometry as described in the text. Results are expressed as percent of control and represent the mean±SD from at least 3 independent experiments. DMSO (solvent) per se did not show any effect on phosphorylation of tested molecules (not shown). Asterisk, p<0.05. (B) EOL-1 cells were incubated in control medium in the absence (Control) or presence of ponatinib, sunitinib, sorafenib, masitinib or erlotinib (each 0.1 or 1 μM) at 37°C for 4 hours. After incubation, cells were analyzed for expression of pPDGFR, pS6 and pSTAT5 by western blotting as described in the text. (C) EOL-1 cells were incubated in control medium (Co) or in various concentrations of everolimus, BEZ235, pimozide, or piceatannol at 37°C for 48 hours. After incubation, 3H-thymidine uptake was measured. Results are expressed as percent of control and represent the mean±SD from 3 independent experiments. Asterisk: p<0.05.
Figure 3. Effects of various targeted drugs on PDGFR-downstream signaling molecules in EOL-1 cells
(A) EOL-1 cells were incubated in control medium in the absence (Co) or presence of ponatinib, sorafenib, masitinib, sunitinib or erlotinib (each 0.01, 0.1 or 1 μM) at 37°C for 4 hours. After incubation, cells were analyzed for expression of pAKT, pS6, and pSTAT5, by flow cytometry as described in the text. Results are expressed as percent of control and represent the mean±SD from at least 3 independent experiments. DMSO (solvent) per se did not show any effect on phosphorylation of tested molecules (not shown). Asterisk, p<0.05. (B) EOL-1 cells were incubated in control medium in the absence (Control) or presence of ponatinib, sunitinib, sorafenib, masitinib or erlotinib (each 0.1 or 1 μM) at 37°C for 4 hours. After incubation, cells were analyzed for expression of pPDGFR, pS6 and pSTAT5 by western blotting as described in the text. (C) EOL-1 cells were incubated in control medium (Co) or in various concentrations of everolimus, BEZ235, pimozide, or piceatannol at 37°C for 48 hours. After incubation, 3H-thymidine uptake was measured. Results are expressed as percent of control and represent the mean±SD from 3 independent experiments. Asterisk: p<0.05.
Figure 3. Effects of various targeted drugs on PDGFR-downstream signaling molecules in EOL-1 cells
(A) EOL-1 cells were incubated in control medium in the absence (Co) or presence of ponatinib, sorafenib, masitinib, sunitinib or erlotinib (each 0.01, 0.1 or 1 μM) at 37°C for 4 hours. After incubation, cells were analyzed for expression of pAKT, pS6, and pSTAT5, by flow cytometry as described in the text. Results are expressed as percent of control and represent the mean±SD from at least 3 independent experiments. DMSO (solvent) per se did not show any effect on phosphorylation of tested molecules (not shown). Asterisk, p<0.05. (B) EOL-1 cells were incubated in control medium in the absence (Control) or presence of ponatinib, sunitinib, sorafenib, masitinib or erlotinib (each 0.1 or 1 μM) at 37°C for 4 hours. After incubation, cells were analyzed for expression of pPDGFR, pS6 and pSTAT5 by western blotting as described in the text. (C) EOL-1 cells were incubated in control medium (Co) or in various concentrations of everolimus, BEZ235, pimozide, or piceatannol at 37°C for 48 hours. After incubation, 3H-thymidine uptake was measured. Results are expressed as percent of control and represent the mean±SD from 3 independent experiments. Asterisk: p<0.05.
Figure 4. Effects of targeted drugs on SDF-1α-induced migration of EOL-1 cells
EOL-1 cells were pre-incubated in control medium (Co) or in medium containing ponatinib, sorafenib, masitinib, nilotinib, imatinib, or dasatinib (each 10 or 100 nM) at 37°C for 1 hour. Then, migration of cells against SDF-1α (30 ng/ml) was determined in a double-chamber chemotaxis assay (4 hours) as described in the text. Numbers of viable migrated cells are expressed as percent of total (100% input) cells. Results represent the mean±S.D. of 4 independent experiments. Asterisk: p<0.05.
Similar articles
- Ponatinib efficiently kills imatinib-resistant chronic eosinophilic leukemia cells harboring gatekeeper mutant T674I FIP1L1-PDGFRα: roles of Mcl-1 and β-catenin.
Jin Y, Ding K, Li H, Xue M, Shi X, Wang C, Pan J. Jin Y, et al. Mol Cancer. 2014 Jan 28;13:17. doi: 10.1186/1476-4598-13-17. Mol Cancer. 2014. PMID: 24472312 Free PMC article. - Activity of AMN107, a novel aminopyrimidine tyrosine kinase inhibitor, against human FIP1L1-PDGFR-alpha-expressing cells.
Verstovsek S, Giles FJ, Quintás-Cardama A, Manshouri T, Huynh L, Manley P, Cortes J, Tefferi A, Kantarjian H. Verstovsek S, et al. Leuk Res. 2006 Dec;30(12):1499-505. doi: 10.1016/j.leukres.2006.03.012. Epub 2006 May 8. Leuk Res. 2006. PMID: 16682077 - Sorafenib is a potent inhibitor of FIP1L1-PDGFRalpha and the imatinib-resistant FIP1L1-PDGFRalpha T674I mutant.
Lierman E, Folens C, Stover EH, Mentens N, Van Miegroet H, Scheers W, Boogaerts M, Vandenberghe P, Marynen P, Cools J. Lierman E, et al. Blood. 2006 Aug 15;108(4):1374-6. doi: 10.1182/blood-2006-02-004457. Epub 2006 Apr 27. Blood. 2006. PMID: 16645167 Free PMC article. - Eosinophilia in mast cell disease.
Kovalszki A, Weller PF. Kovalszki A, et al. Immunol Allergy Clin North Am. 2014 May;34(2):357-64. doi: 10.1016/j.iac.2014.01.013. Epub 2014 Mar 13. Immunol Allergy Clin North Am. 2014. PMID: 24745679 Free PMC article. Review. - FIP1L1/PDGFR alpha-associated systemic mastocytosis.
Yamada Y, Cancelas JA. Yamada Y, et al. Int Arch Allergy Immunol. 2010;152 Suppl 1(Suppl 1):101-5. doi: 10.1159/000312134. Epub 2010 Jun 4. Int Arch Allergy Immunol. 2010. PMID: 20523072 Free PMC article. Review.
Cited by
- CLIP-170S is a microtubule +TIP variant that confers resistance to taxanes by impairing drug-target engagement.
Thakkar PV, Kita K, Castillo UD, Galletti G, Madhukar N, Navarro EV, Barasoain I, Goodson HV, Sackett D, Díaz JF, Lu Y, RoyChoudhury A, Molina H, Elemento O, Shah MA, Giannakakou P. Thakkar PV, et al. Dev Cell. 2021 Dec 6;56(23):3264-3275.e7. doi: 10.1016/j.devcel.2021.09.023. Epub 2021 Oct 20. Dev Cell. 2021. PMID: 34672971 Free PMC article. - Eosinophils and eosinophil-associated disorders: immunological, clinical, and molecular complexity.
Valent P, Degenfeld-Schonburg L, Sadovnik I, Horny HP, Arock M, Simon HU, Reiter A, Bochner BS. Valent P, et al. Semin Immunopathol. 2021 Jun;43(3):423-438. doi: 10.1007/s00281-021-00863-y. Epub 2021 May 30. Semin Immunopathol. 2021. PMID: 34052871 Free PMC article. Review. - Zebrafish as a Model for In-Depth Mechanistic Study for Stroke.
Chen W, Xie L, Yu F, Li Y, Chen C, Xie W, Huang T, Zhang Y, Zhang S, Li P. Chen W, et al. Transl Stroke Res. 2021 Oct;12(5):695-710. doi: 10.1007/s12975-021-00907-3. Epub 2021 May 29. Transl Stroke Res. 2021. PMID: 34050491 Review. - Myeloid Neoplasm with PDGFRA Rearrangement Manifesting as a Retromolar Pad Mass.
Duffield AS, Webster J, Smith BD, Necciai JS, McCuiston A, Ware AD. Duffield AS, et al. Head Neck Pathol. 2021 Dec;15(4):1399-1403. doi: 10.1007/s12105-021-01305-9. Epub 2021 Feb 22. Head Neck Pathol. 2021. PMID: 33616851 Free PMC article. - S100A8 and S100A9 Promote Apoptosis of Chronic Eosinophilic Leukemia Cells.
Lee JS, Lee NR, Kashif A, Yang SJ, Nam AR, Song IC, Gong SJ, Hong MH, Kim G, Seok PR, Lee MS, Sung KH, Kim IS. Lee JS, et al. Front Immunol. 2020 Aug 6;11:1258. doi: 10.3389/fimmu.2020.01258. eCollection 2020. Front Immunol. 2020. PMID: 32903598 Free PMC article.
References
- Gotlib J. Molecular classification and pathogenesis of eosinophilic disorders: 2005 update. Acta Haematol. 2005;114:7–25. - PubMed
- Tefferi A, Patnaik MM, Pardanani A. Eosinophilia: secondary, clonal and idiopathic. Br J Haematol. 2006 Jun;133:468–492. - PubMed
- Bain BJ, Fletcher SH. Chronic eosinophilic leukemias and the myeloproliferative variant of the hypereosinophilic syndrome. Immunol Allergy Clin North Am. 2007;27:377–388. - PubMed
- Gotlib J. Chronic eosinophilic leukemia/hypereosinophilic syndrome. Cancer Treat Res. 2008;142:69–106. - PubMed
- Valent P. Pathogenesis, classification, and therapy of eosinophilia and eosinophil disorders. Blood Rev. 2009;23:157–165. - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Research Materials
Miscellaneous