Combination of the ABL kinase inhibitor imatinib with the Janus kinase 2 inhibitor TG101348 for targeting residual BCR-ABL-positive cells

Journal of Hematology & Oncology

Kazuma Ohyashiki¹  Yuko Tanaka¹  Seiichiro Katagiri¹  Tetsuzo Tauchi¹  Seiichi Okabe¹ 

[1] First Department of Internal Medicine, Tokyo Medical University, Shinjuku-ku, Tokyo 160-0023, Japan

关键词: Feeder cell;    Cytokine;    Imatinib;    JAK2 inhibitor;    Chronic myeloid leukemia;   

Others  :  801234
DOI  :  10.1186/1756-8722-7-37

 received in 2014-02-07, accepted in 2014-04-22,  发布年份 2014

【 摘 要 】

Background

The ABL kinase inhibitor imatinib is highly effective in treating most, but not all, patients with chronic myeloid leukemia (CML). This is because residual CML cells are generally present in the bone marrow microenvironment and are refractory to imatinib. Hematopoietic cytokine receptor signaling is mediated by Janus kinases (JAKs) and their downstream transcription factor, signal transducer and activator of transcription (STAT). TG101348 (SAR302503) is an oral inhibitor of JAK2.

Methods

We investigated the efficacy of imatinib and TG101348 using the break point cluster region-c-Abelson (BCR-ABL)-positive cell line and primary CML samples wherein leukemia cells were protected by a feeder cell line (HS-5).

Results

Imatinib treatment resulted in partial inhibition of cell growth in HS-5-conditioned medium. Furthermore, combined treatment with imatinib and TG101348 abrogated the protective effects of HS-5-conditioned medium on K562 cells. Phosphorylation of Crk-L, a BCR-ABL substrate, decreased considerably, while apoptosis increased. In addition, the combined treatment of CD34-positive primary samples resulted in considerably increased cytotoxicity, decreased Crk-L phosphorylation, and increased apoptosis. We also investigated TG101348 activity against feeder cells and observed that STAT5 phosphorylation, granulocyte macrophage colony-stimulating factor, and interleukin 6 levels decreased, indicating reduced cytokine production in HS-5 cells treated with TG101348.

Conclusions

These results showed that JAK inhibitors may enhance the cytotoxic effect of imatinib against residual CML cells and that a combined approach may be a powerful strategy against the stroma-associated drug resistance of Philadelphia chromosome-positive cells.

【 授权许可】

   
2014 Okabe et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Sattler M, Griffin JD: Molecular mechanisms of transformation by the BCR-ABL oncogene. Semin Hematol 2003, 40(suppl 1):4-10.
• [2]Kantarjian HM, Talpaz M, Giles F, O’Brien S, Cortes J: New insights into the pathophysiology of chronic myeloid leukemia and imatinib resistance. Ann Intern Med 2006, 145:913-923.
• [3]Helgason GV, Karvela M, Holyoake TL: Kill one bird with two stones: potential efficacy of BCR-ABL and autophagy inhibition in CML. Blood 2011, 118:2035-2043.
• [4]Sawyers CL, McLaughlin J, Witte ON: Genetic requirement for Ras in the transformation of fibroblasts and hematopoietic cells by the Bcr-Abl oncogene. J Exp Med 1995, 181:307-313.
• [5]Skorski T, Bellacosa A, Nieborowska-Skorska M, Majewski M, Martinez R, Choi JK, Trotta R, Wlodarski P, Perrotti D, Chan TO, Wasik MA, Tsichlis PN, Calabretta B: Transformation of hematopoietic cells by BCR/ABL requires activation of a PI-3k/Akt-dependent pathway. EMBO J 1997, 16:6151-6161.
• [6]Chai SK, Nichols GL, Rothman P: Constitutive activation of JAKs and STATs in BCR-Abl-expressing cell lines and peripheral blood cells derived from leukemic patients. J Immunol 1997, 159:4720-4728.
• [7]Roskoski R Jr: STI-571: an anticancer protein-tyrosine kinase inhibitor. Biochem Biophys Res Commun 2003, 309:709-717.
• [8]Druker BJ, Guilhot F, O’Brien SG, Gathmann I, Kantarjian H, Gattermann N, Deininger MW, Silver RT, Goldman JM, Stone RM, Cervantes F, Hochhaus A, Powell BL, Gabrilove JL, Rousselot P, Reiffers J, Cornelissen JJ, Hughes T, Agis H, Fischer T, Verhoef G, Shepherd J, Saglio G, Gratwohl A, Nielsen JL, Radich JP, Simonsson B, Taylor K, Baccarani M, So C, et al.: Five-year follow-up of patients receiving imatinib for chronic myeloid leukemia. N Engl J Med 2006, 355:2408-2417.
• [9]Mahon FX, Réa D, Guilhot J, Guilhot F, Huguet F, Nicolini F, Legros L, Charbonnier A, Guerci A, Varet B, Etienne G, Reiffers J, Rousselot P, Intergroupe Français des Leucémies Myéloïdes Chroniques: Discontinuation of imatinib in patients with chronic myeloid leukemia who have maintained complete molecular remission for at least 2 years: the prospective, multicentre Stop Imatinib (STIM) trial. Lancet Oncol 2010, 11:1029-1035.
• [10]Sloma I, Jiang X, Eaves AC, Eaves CJ: Insights into the stem cells of chronic myeloid leukemia. Leukemia 2010, 24:1823-1833.
• [11]Murray PJ: The JAK-STAT signaling pathway: input and output integration. J Immunol 2007, 178:2623-2629.
• [12]Wheadon H, Roberts PJ, Watts MJ, Linch DC: Changes in signal transduction downstream from the granulocyte-macrophage colony-stimulating factor receptor during differentiation of primary hemopoietic cells. Exp Hematol 1999, 27:1077-1086.
• [13]Nagata Y, Todokoro K: Interleukin 3 activates not only JAK2 and STAT5, but also Tyk2, STAT1, and STAT3. Biochem Biophys Res Commun 1996, 221:785-789.
• [14]Fabbro D: BCR-ABL signaling: A new STATus in CML. Nat Chem Biol 2012, 8:228-229.
• [15]Vainchenker W, Constantinescu SN: JAK/STAT signaling in hematological malignancies. Oncogene 2013, 32:2601-2613.
• [16]Pardanani A, Gotlib JR, Jamieson C, Cortes JE, Talpaz M, Stone RM, Silverman MH, Gilliland DG, Shorr J, Tefferi A: Safety and efficacy of TG101348, a selective JAK2 inhibitor, in myelofibrosis. J Clin Oncol 2011, 29:789-796.
• [17]Warsch W, Walz C, Sexl V: JAK of all trades: JAK2-STAT5 as novel therapeutic targets in BCR-ABL1+ chronic myeloid leukemia. Blood 2013, 122:2167-2175.
• [18]Xu W, Chen B, Tong X: Chronic myeloid leukemia patient with co-occurrence of BCR-ABL junction and JAK2 V617F mutation. Int J Hematol 2014, 99:87-90.
• [19]Lee YJ, Moon JH, Shin HC, Seo JW, Han SA, Seo SK, Sohn SK: Two CML patients who subsequently developed features of essential thrombocythemia with JAK2-V617F mutation while in complete cytogenetic remission after treatment with imatinib mesylate. Int J Hematol 2013, 97:804-807.
• [20]Park SH, Chi HS, Cho YU, Jang S, Park CJ, Kim DY, Lee JH, Lee KH: Two cases of myeloproliferative neoplasm with a concurrent JAK2 (V617F) mutation and BCR/ABL translocation without chronic myelogenous leukemia phenotype acquisition during hydroxyurea treatment. Ann Lab Med 2013, 33:229-232.
• [21]Samanta AK, Chakraborty SN, Wang Y, Schlette E, Reddy EP, Arlinghaus RB: Destabilization of Bcr-Abl/Jak2 network by a Jak2/Abl kinase inhibitor ON044580 overcomes drug resistance in blast crisis chronic myelogenous leukemia (CML). Genes Cancer 2010, 1:346-359.
• [22]Cogle CR: Overcoming chronic myeloid leukemia stem cell resistance to imatinib by also targeting JAK2. J Natl Cancer Inst 2013, 105:378-379.
• [23]Nakao T, Kim S, Ohta K, Kawano H, Hino M, Miura K, Tatsumi N, Iwao H: Role of mitogen-activated protein kinase family in serum-induced leukemia inhibitory factor and interleukin-6 secretion by bone marrow stromal cells. Br J Pharmacol 2002, 136:975-984.
• [24]Ernst T, Hochhaus A: Chronic myeloid leukemia: clinical impact of BCR-ABL1 mutations and other lesions associated with disease progression. Semin Oncol 2012, 39:58-66.
• [25]Warsch W, Kollmann K, Eckelhart E, Fajmann S, Cerny-Reiterer S, Hölbl A, Gleixner KV, Dworzak M, Mayerhofer M, Hoermann G, Herrmann H, Sillaber C, Egger G, Valent P, Moriggl R, Sexl V: High STAT5 levels mediate imatinib resistance and indicate disease progression in chronic myeloid leukemia. Blood 2011, 117:3409-3420.
• [26]Corbin AS, Agarwal A, Loriaux M, Cortes J, Deininger MW, Druker BJ: Human chronic myeloid leukemia stem cells are insensitive to imatinib despite inhibition of BCR-ABL activity. J Clin Invest 2011, 121:396-409.
• [27]Wang Y, Cai D, Brendel C, Barett C, Erben P, Manley PW, Hochhaus A, Neubauer A, Burchert A: Adaptive secretion of granulocyte-macrophage colony-stimulating factor (GM-CSF) mediates imatinib and nilotinib resistance in BCR/ABL + progenitors via JAK-2/STAT-5 pathway activation. Blood 2007, 109:2147-2155.
• [28]Mohamed AA, Ibrahim AM, El-Masry MW, Mansour IM, Khroshied MA, Gouda HM, Riad RM: Ex vivo expansion of stem cells: defining optimum conditions using various cytokines. Lab Hematol 2006, 12:86-93.
• [29]McCubrey JA, Steelman LS, Abrams SL, Bertrand FE, Ludwig DE, Bäsecke J, Libra M, Stivala F, Milella M, Tafuri A, Lunghi P, Bonati A, Martelli AM: Targeting survival cascades induced by activation of Ras/Raf/MEK/ERK, PI3K/PTEN/Akt/mTOR and Jak/STAT pathways for effective leukemia therapy. Leukemia 2008, 22:708-722.
• [30]Okabe S, Tauchi T, Tanaka Y, Katagiri S, Ohyashiki K: Effects of the hedgehog inhibitor GDC-0449, alone or in combination with dasatinib, on BCR-ABL-positive leukemia cells. Stem Cells Dev 2012, 21:2939-2948.
• [31]Okabe S, Tauchi T, Tanaka Y, Ohyashiki K: Efficacy of ponatinib against ABL tyrosine kinase inhibitor-resistant leukemia cells. Biochem Biophys Res Commun 2013, 435:506-511.