Advertisement

International Journal of Hematology

, Volume 110, Issue 2, pp 213–227 | Cite as

Binimetinib, a novel MEK1/2 inhibitor, exerts anti-leukemic effects under inactive status of PI3Kinase/Akt pathway

  • Kanae Sakakibara
  • Takayuki Tsujioka
  • Jun-ichiro Kida
  • Nami Kurozumi
  • Takako Nakahara
  • Shin-ichiro Suemori
  • Akira Kitanaka
  • Yujiro Arao
  • Kaoru TohyamaEmail author
Original Article
  • 104 Downloads

Abstract

A MEK1/2 inhibitor, binimetinib is promising as a therapeutic agent for malignant melanoma with N-RAS mutation. We examined in vitro effects of binimetinib on 10 human myeloid/lymphoid leukemia cell lines, and found that three of five cell lines with N-RAS mutation and one of five without N-RAS mutation were responsive to treatment with binimetinib. Binimetinib inhibited cell growth mainly by inducing G1 arrest and this action mechanism was assisted by gene set enrichment analysis. To identify signaling pathways associated with binimetinib response, we examined the status of MAP kinase/ERK and PI3Kinase/Akt pathways. The basal levels of phosphorylated ERK and Akt varied between the cell lines, and the amounts of phosphorylated ERK and Akt appeared to be reciprocal of each other. Interestingly, most of the binimetinib-resistant cell lines revealed strong Akt phosphorylation compared with binimetinib-sensitive ones. The effect of binimetinib may not be predicted by the presence/absence of N-RAS mutation, but rather by Akt phosphorylation status. Moreover, combination of binimetinib with a PI3K/Akt inhibitor showed additive growth-suppressive effects. These results suggest that binimetinib shows potential anti-leukemic effects and the basal level of phosphorylated Akt might serve as a biomarker predictive of therapeutic effect.

Keywords

Myelodysplastic syndromes (MDS) N-RAS mutation G1 arrest Akt phosphorylation 

Notes

Acknowledgements

This work was supported in part by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science, and in part by a Kawasaki Medical School project grant. The authors thank Ms. Aki Kuyama for editorial assistance.

Compliance with ethical standards

Conflict of interest

The authors have no conflict of interest to declare.

Supplementary material

12185_2019_2667_MOESM1_ESM.jpg (942 kb)
Binimetinib inhibits the proliferation of some leukemia cell lines. Five cell lines with N-RAS mutation (HL-60, MDS-L, TF-1, THP-1 and MOLT4) and five cell lines without this mutation (Jurkat, K562, U937, MOLM13 and F-36P) were treated with binimetinib (0-1 µM) for indicated times (24, 48, 72 and 96 h) and cell growth was assessed by trypan blue staining. The value without binimetinib was adjusted to 100%. The data represent the mean values with SD from three independent experiments (JPEG 941 kb)
12185_2019_2667_MOESM2_ESM.jpg (147 kb)
Buparlisib inhibits the proliferation of TF-1 and F-36P cell lines. TF-1 and F-36P were treated with 1 µM buparlisib for 48 h and cell count was evaluated by trypan blue staining (JPEG 146 kb)

References

  1. 1.
    Miyawaki S. JSH guideline for tumors of hematopoietic and lymphoid tissues: leukemia 1. Acute myeloid leukemia (AML). Int J Hematol. 2017;106:310–25.CrossRefGoogle Scholar
  2. 2.
    Takeuchi J, Kusumoto S, Akiyama H, Kanda Y, Izutsu K. JSH guideline for tumors of hematopoietic and lymphoid tissues-leukemia: 3. Acute lymphoblastic leukemia/lymphoblastic lymphoma (ALL/LBL). Int J Hematol. 2017;106:732–47.CrossRefGoogle Scholar
  3. 3.
    Prior IA, Lewis PD, Mattos C. A comprehensive survey of Ras mutations in cancer. Cancer Res. 2012;72:2457–67.CrossRefGoogle Scholar
  4. 4.
    Cazzola M, Della Porta MG, Malcovati L. The genetic basis of myelodysplasia and its clinical relevance. Blood. 2013;122:4021–34.CrossRefGoogle Scholar
  5. 5.
    Welch JS, Ley TJ, Link DC, Miller CA, Larson DE, Koboldt DC, et al. The origin and evolution of mutations in acute myeloid leukemia. Cell. 2012;150:264–78.CrossRefGoogle Scholar
  6. 6.
    Bacher U, Haferlach T, Kern W, Haferlach C, Schnittger S. A comparative study of molecular mutations in 381 patients with myelodysplastic syndrome and in 4130 patients with acute myeloid leukemia. Haematologica. 2007;92:744–52.CrossRefGoogle Scholar
  7. 7.
    Patel JP, Gonen M, Figueroa ME, Fernandez H, Sun Z, Racevskis J, et al. Prognostic relevance of integrated genetic profiling in acute myeloid leukemia. N Engl J Med. 2012;366:1079–89.CrossRefGoogle Scholar
  8. 8.
    Christiansen DH, Andersen MK, Desta F, Pedersen-Bjergaard J. Mutations of genes in the receptor tyrosine kinase (RTK)/RAS-BRAF signal transduction pathway in therapy-related myelodysplasia and acute myeloid leukemia. Leukemia. 2005;19:2232–40.CrossRefGoogle Scholar
  9. 9.
    Miller CR, Oliver KE, Farley JH. MEK1/2 inhibitors in the treatment of gynecologic malignancies. Gynecol Oncol. 2014;133:128–37.CrossRefGoogle Scholar
  10. 10.
    Akinleye A, Furqan M, Mukhi N, Ravella P, Liu D. MEK and the inhibitors: from bench to bedside. J Hematol Oncol. 2013;6:27.CrossRefGoogle Scholar
  11. 11.
    Yao W, Yue P, Zhang G, Owonikoko TK, Khuri FR, Sun SY. Enhancing therapeutic efficacy of the MEK inhibitor, MEK162, by blocking autophagy or inhibiting PI3K/Akt signaling in human lung cancer cells. Cancer Lett. 2015;364:70–8.CrossRefGoogle Scholar
  12. 12.
    Hamidi H, Lu M, Chau K, Anderson L, Fejzo M, Ginther C, et al. KRAS mutational subtype and copy number predict in vitro response of human pancreatic cancer cell lines to MEK inhibition. Br J Cancer. 2014;111:1788–801.CrossRefGoogle Scholar
  13. 13.
    Lee MS, Helms TL, Feng N, Gay J, Chang QE, Tian F, et al. Efficacy of the combination of MEK and CDK4/6 inhibitors in vitro and in vivo in KRAS mutant colorectal cancer models. Oncotarget. 2016;7:39595–608.Google Scholar
  14. 14.
    Kiessling MK, Curioni-Fontecedro A, Samaras P, Lang S, Scharl M, Aguzzi A, et al. Targeting the mTOR complex by everolimus in NRAS mutant neuroblastoma. PLoS One. 2016;11:e0147682.CrossRefGoogle Scholar
  15. 15.
    Thumar J, Shahbazian D, Aziz SA, Jilaveanu LB, Kluger HM. MEK targeting in N-RAS mutated metastatic melanoma. Mol Cancer. 2014;13:45.CrossRefGoogle Scholar
  16. 16.
    Kerstjens M, Driessen EM, Willekes M, Pinhancos SS, Schneider P, Pieters R, et al. MEK inhibition is a promising therapeutic strategy for MLL-rearranged infant acute lymphoblastic leukemia patients carrying RAS mutations. Oncotarget. 2017;8:14835–46.CrossRefGoogle Scholar
  17. 17.
    Matsuoka A, Tochigi A, Kishimoto M, Nakahara T, Kondo T, Tsujioka T, et al. Lenalidomide induces cell death in an MDS-derived cell line with deletion of chromosome 5q by inhibition of cytokinesis. Leukemia. 2010;24:748–55.CrossRefGoogle Scholar
  18. 18.
    Tsujioka T, Yokoi A, Uesugi M, Kishimoto M, Tochigi A, Suemori S, et al. Effects of DNA methyltransferase inhibitors (DNMTIs) on MDS-derived cell lines. Exp Hematol. 2013;41:189–97.CrossRefGoogle Scholar
  19. 19.
    Kida JI, Tsujioka T, Suemori SI, Okamoto S, Sakakibara K, Takahata T, et al. An MDS-derived cell line and a series of its sublines serve as an in vitro model for the leukemic evolution of MDS. Leukemia. 2018;32:1846–50.CrossRefGoogle Scholar
  20. 20.
    Ikediobi ON, Davies H, Bignell G, Edkins S, Stevens C, O’Meara S, et al. Mutation analysis of 24 known cancer genes in the NCI-60 cell line set. Mol Cancer Ther. 2006;5:2606–12.CrossRefGoogle Scholar
  21. 21.
    Yu Y, Xie Y, Cao L, Yang L, Yang M, Lotze MT, et al. The ferroptosis inducer erastin enhances sensitivity of acute myeloid leukemia cells to chemotherapeutic agents. Mol Cell Oncol. 2015;2:e1054549.CrossRefGoogle Scholar
  22. 22.
    Ehmann F, Horn S, Garcia-Palma L, Wegner W, Fiedler W, Giehl K, et al. Detection of N-RAS and K-RAS in their active GTP-bound form in acute myeloid leukemia without activating RAS mutations. Leuk Lymphoma. 2006;47:1387–91.CrossRefGoogle Scholar
  23. 23.
    Cancer Cell Line Encyclopedia. https://portals.broadinstitute.org/ccle. Accessed 24 April 2019.
  24. 24.
    Ormerod MG, Collins MK, Rodriguez-Tarduchy G, Robertson D. Apoptosis in interleukin-3-dependent haemopoietic cells. Quantification by two flow cytometric methods. J Immunol Methods. 1992;153:57–65.CrossRefGoogle Scholar
  25. 25.
    Vermes I, Haanen C, Steffens-Nakken H, Reutelingsperger C. A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. J Immunol Methods. 1995;184:39–51.CrossRefGoogle Scholar
  26. 26.
    Morita H, Matsuoka A, Kida JI, Tabata H, Tohyama K, Tohyama Y. KIF20A, highly expressed in immature hematopoietic cells, supports the growth of HL60 cell line. Int J Hematol. 2018;108:607–14.CrossRefGoogle Scholar
  27. 27.
    Shi Y, Tohyama Y, Kadono T, He J, Miah SM, Hazama R, et al. Protein-tyrosine kinase Syk is required for pathogen engulfment in complement-mediated phagocytosis. Blood. 2006;107:4554–62.CrossRefGoogle Scholar
  28. 28.
    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;102:15545–50.CrossRefGoogle Scholar
  29. 29.
    Gene set enrichment analysis: http://software.broadinstitute.org/gsea/index.jsp. Accessed 24 April 2019.
  30. 30.
    Garon EB, Finn RS, Hosmer W, Dering J, Ginther C, Adhami S, et al. Identification of common predictive markers of in vitro response to the Mek inhibitor selumetinib (AZD6244; ARRY-142886) in human breast cancer and non-small cell lung cancer cell lines. Mol Cancer Ther. 2010;9:1985–94.CrossRefGoogle Scholar
  31. 31.
    Watanabe K, Otsu S, Hirashima Y, Morinaga R, Nishikawa K, Hisamatsu Y, et al. A phase I study of binimetinib (MEK162) in Japanese patients with advanced solid tumors. Cancer Chemother Pharmacol. 2016;77:1157–64.CrossRefGoogle Scholar
  32. 32.
    Bendell JC, Javle M, Bekaii-Saab TS, Finn RS, Wainberg ZA, Laheru DA, et al. A phase 1 dose-escalation and expansion study of binimetinib (MEK162), a potent and selective oral MEK1/2 inhibitor. Br J Cancer. 2017;116:575–83.CrossRefGoogle Scholar
  33. 33.
    Balmanno K, Chell SD, Gillings AS, Hayat S, Cook SJ. Intrinsic resistance to the MEK1/2 inhibitor AZD6244 (ARRY-142886) is associated with weak ERK1/2 signalling and/or strong PI3K signalling in colorectal cancer cell lines. Int J Cancer. 2009;125:2332–41.CrossRefGoogle Scholar
  34. 34.
    Yeh JJ, Routh ED, Rubinas T, Peacock J, Martin TD, Shen XJ, et al. KRAS/BRAF mutation status and ERK1/2 activation as biomarkers for MEK1/2 inhibitor therapy in colorectal cancer. Mol Cancer Ther. 2009;8:834–43.CrossRefGoogle Scholar
  35. 35.
    Wan X, Helman LJ. Levels of PTEN protein modulate Akt phosphorylation on serine 473, but not on threonine 308, in IGF-II-overexpressing rhabdomyosarcomas cells. Oncogene. 2003;22:8205–11.CrossRefGoogle Scholar
  36. 36.
    Tesio M, Trinquand A, Ballerini P, Hypolite G, Lhermitte L, Petit A, et al. Age-related clinical and biological features of PTEN abnormalities in T-cell acute lymphoblastic leukaemia. Leukemia. 2017;31:2594–600.CrossRefGoogle Scholar
  37. 37.
    He W, Wang X, Chen L, Guan X. A crosstalk imbalance between p27(Kip1) and its interacting molecules enhances breast carcinogenesis. Cancer Biother Radiopharm. 2012;27:399–402.CrossRefGoogle Scholar
  38. 38.
    Timmerbeul I, Garrett-Engele CM, Kossatz U, Chen X, Firpo E, Grunwald V, et al. Testing the importance of p27 degradation by the SCFskp2 pathway in murine models of lung and colon cancer. Proc Natl Acad Sci USA. 2006;103:14009–14.CrossRefGoogle Scholar
  39. 39.
    Ungermannova D, Gao Y, Liu X. Ubiquitination of p27Kip1 requires physical interaction with cyclin E and probable phosphate recognition by SKP2. J Biol Chem. 2005;280:30301–9.CrossRefGoogle Scholar
  40. 40.
    Chen G, Li G. Increased Cul1 expression promotes melanoma cell proliferation through regulating p27 expression. Int J Oncol. 2010;37:1339–44.Google Scholar
  41. 41.
    Bai J, Zhou Y, Chen G, Zeng J, Ding J, Tan Y, et al. Overexpression of Cullin1 is associated with poor prognosis of patients with gastric cancer. Hum Pathol. 2011;42:375–83.CrossRefGoogle Scholar
  42. 42.
    Min KW, Kim DH, Do SI, Sohn JH, Chae SW, Pyo JS, et al. Diagnostic and prognostic relevance of Cullin1 expression in invasive ductal carcinoma of the breast. J Clin Pathol. 2012;65:896–901.CrossRefGoogle Scholar
  43. 43.
    Taskinen M, Louhimo R, Koivula S, Chen P, Rantanen V, Holte H, et al. Deregulation of COMMD1 is associated with poor prognosis in diffuse large B-cell lymphoma. PLoS One. 2014;9:e91031.CrossRefGoogle Scholar
  44. 44.
    Malek E, Abdel-Malek MA, Jagannathan S, Vad N, Karns R, Jegga AG, et al. Pharmacogenomics and chemical library screens reveal a novel SCF(SKP2) inhibitor that overcomes Bortezomib resistance in multiple myeloma. Leukemia. 2017;31:645–53.CrossRefGoogle Scholar
  45. 45.
    Lonetti A, Antunes IL, Chiarini F, Orsini E, Buontempo F, Ricci F, et al. Activity of the pan-class I phosphoinositide 3-kinase inhibitor NVP-BKM120 in T-cell acute lymphoblastic leukemia. Leukemia. 2014;28:1196–206.CrossRefGoogle Scholar
  46. 46.
    Turke AB, Song Y, Costa C, Cook R, Arteaga CL, Asara JM, et al. MEK inhibition leads to PI3K/AKT activation by relieving a negative feedback on ERBB receptors. Cancer Res. 2012;72:3228–37.CrossRefGoogle Scholar
  47. 47.
    Gopal YN, Deng W, Woodman SE, Komurov K, Ram P, Smith PD, et al. Basal and treatment-induced activation of AKT mediates resistance to cell death by AZD6244 (ARRY-142886) in Braf-mutant human cutaneous melanoma cells. Cancer Res. 2010;70:8736–47.CrossRefGoogle Scholar

Copyright information

© Japanese Society of Hematology 2019

Authors and Affiliations

  1. 1.Division of Medical TechnologyKawasaki University of Medical WelfareOkayamaJapan
  2. 2.Field of Medical Technology, Graduate School of Health SciencesOkayama UniversityOkayamaJapan
  3. 3.Department of Laboratory MedicineKawasaki Medical SchoolKurashikiJapan

Personalised recommendations