SNS-032 inhibits mTORC1/mTORC2 activity in acute myeloid leukemia cells and has synergistic activity with perifosine against Akt

Journal of Hematology & Oncology

Wenbin Qian¹  Xue Yang¹  Chunmei Yang¹  Liangshun You¹  Hui Liu¹  Yingming Jin¹  Haitao Meng¹ 

[1] Institute of Hematology, The First Affiliated Hospital, College of Medicine, Zhejiang University, 79# Qingchun Road, Hangzhou, 310003, P. R. China

关键词: Acute myeloid leukemia;    Akt;    Perifosine;    Cyclin-dependent kinases;    mTORC2;    mTORC1;    SNS-032;   

Others  :  812547
DOI  :  10.1186/1756-8722-6-18

 received in 2012-11-18, accepted in 2013-02-08,  发布年份 2013

【 摘 要 】

Background

Acute myeloid leukemia (AML) is a heterogeneous disorder with aberrant regulation of a variety of signal pathways. Therefore, simultaneous targeting of two or even more deregulated signal transduction pathways is needed to overcome drug resistance. Previously, it was reported that SNS-032, a selective cyclin-dependent kinase inhibitor, is an effective agent for treatment of AML; however, the molecular mechanisms of SNS-032-induced cell death of AML cells are not yet fully understood. The aim of the study was to characterize the effects in vitro of SNS-032, used alone and in combination with an Akt inhibitor perifosine, against AML cells and to identify the mechanism involved.

Results

SNS-032 significantly induced cytotoxicity in human AML cell lines and blasts from patients with newly diagnosed or relapsed AML. However, Kasumi-1 cells and some of leukemic samples (14.9%) from AML patients were resistant to SNS-032-mediated cell death. Western blot analysis showed that SNS-032 strongly inhibited the phosphorylation of mammalian target of rapamycin (mTOR) on Ser 2448 and Ser2481, and that removal of SNS-032 resulted in partial recovery of cell death and reactivation of phosphorylation of mTOR. Moreover, exogenous insulin-like growth factor-1 (IGF-1) did not reverse SNS-032-induced cell growth inhibition and downregualtion of phosphor-mTOR at Ser2448 and Ser2481 although slight suppression of IGF-1R expression was triggered by the agent. Furthermore, SNS-032 at a lower concentration (60–80 nM) enhanced AML cell cytotoxicity induced by perifosine, an Akt inhibitor. Importantly, SNS-032 treatment reduced colony formation ability of AML cells, which was significantly increased when two agents were combined. This combination therapy led to almost complete inhibition of Akt activity.

Conclusion

We conclude that SNS-032 might directly target mammalian target of rapamycin complex 1 (mTORC1)/mTORC2. Our results further provide a rationale for combining SNS-032 with perifosine for the treatment of AML.

【 授权许可】

   
2013 Meng et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

Figure 7.

【 参考文献 】

• [1]Burnett AK, Hills RK, Milligan DW, Goldstone AH, Prentice AG, McMullin MF, Duncombe A, Gibson B, Wheatley K: Attempts to optimize induction and consolidation treatment in acute myeloid leukemia: results of the MRC AML12 trial. J Clin Oncol 2010, 28:586-595.
• [2]Büchner T, Berdel WE, Haferlach C, Haferlach T, Schnittger S, Müller-Tidow C, Braess J, Spiekermann K, Kienast J, Staib P: Age-related risk profile and chemotherapy dose response in acute myeloid leukemia: a study by the German acute myeloid leukemia cooperative group. J Clin Oncol 2009, 27:61-69.
• [3]Scholl C, Gilliland DG, Fröhling S: Deregulation of signaling pathways in acute myeloid leukemia. Semin Oncol 2008, 35:336-345.
• [4]Zhu X, Ma Y, Liu D: Novel agents and regimens for acute myeloid leukemia: 2009 ASH annual meeting highlights. J Hematol Oncol 2010, 23:3-17.
• [5]Roboz GJ: Novel approaches to the treatment of acute myeloid leukemia. Hematology Am Soc Hematol Educ Program 2011, 2011:43-50.
• [6]Senderowicz AM: Inhibitors of cyclin-dependent kinase modulators for cancer therapy. Prog Drug Res 2005, 63:183-206.
• [7]Dickson MA, Schwartz GK: Development of cell-cycle inhibitors for cancer therapy. Curr Oncol 2009, 16:36-43.
• [8]Chen R, Wierda WG, Chubb S, Hawtin RE, Fox JA, Keating MJ, Gandhi V, Plunkett W: Mechanism of action of SNS-032, a novel cyclin-dependent kinase inhibitor, in chronic lymphocytic leukemia. Blood 2009, 113:4637-4645.
• [9]Chen R, Chubb S, Cheng T, Hawtin RE, Gandhi V, Plunkett W: Responses in mantle cell lymphoma cells to SNS-032 depend on the biological context of each cell line. Cancer Res 2010, 70:6587-6597.
• [10]Wu Y, Chen C, Sun X, Shi X, Jin B, Ding K, Yeung SC, Pan J: Cyclin-dependent kinase 7/9 inhibitor SNS-032 abrogates FIP1-like-1 platelet-derived growth factor receptor α and bcr-abl oncogene addiction in malignant hematologic cells. Clin Cancer Res 2012, 18:1966-1978.
• [11]Tong WG, Chen R, Plunkett W, Siegel D, Sinha R, Harvey RD, Badros AZ, Popplewell L, Coutre S, Fox JA: Phase I and pharmacologic study of SNS-032, a potent and selective Cdk2, 7, and 9 inhibitor, in patients with advanced chronic lymphocytic leukemia and multiple myeloma. J Clin Oncol 2010, 28:3015-3022.
• [12]Walsby E, Lazenby M, Pepper C, Burnett AK: The cyclin-dependent kinase inhibitor SNS-032 has single agent activity in AML cells and is highly synergistic with cytarabine. Leukemia 2011, 25:411-419.
• [13]Park S, Chapuis N, Tamburini J, Bardet V, Cornillet-Lefebvre P, Willems L, Green A, Mayeux P, Lacombe C, Bouscary D: Role of the PI3K/AKT and mTOR signaling pathways in acute myeloid leukemia. Haematologica 2010, 95:819-828.
• [14]Oh WJ, Jacinto E: mTOR complex 2 signaling and functions. Cell Cycle 2011, 10:2305-2316.
• [15]Chapuis N, Tamburini J, Green AS, Vignon C, Bardet V, Neyret A, Pannetier M, Willems L, Park S, Macone A: Dual inhibition of PI3K and mTORC1/2 signaling by NVP-BEZ235 as a New therapeutic strategy for acute myeloid leukemia. Clin Cancer Res 2010, 16:5424-5435.
• [16]Gupta M, Hendrickson AE, Yun SS, Han JJ, Schneider PA, Koh BD, Stenson MJ, Wellik LE, Shing JC, Peterson KL: Dual mTORC1/mTORC2 inhibition diminishes Akt activation and induces Puma-dependent apoptosis in lymphoid malignancies. Blood 2012, 119:476-487.
• [17]Zeng Z, Shi YX, Tsao T, Qiu Y, Kornblau SM, Baggerly KA, Liu W, Jessen K, Liu Y, Kantarjian H: Targeting of mTORC1/2 by the mTOR kinase inhibitor PP242 induces apoptosis in AML cells under conditions mimicking the bone marrow microenvironment. Blood 2012, 120:2679-2689.
• [18]Wahner Hendrickson AE, Haluska P, Schneider PA, Loegering DA, Peterson KL, Attar R, Smith BD, Erlichman C, Gottardis M, Karp JE: Expression of insulin receptor isoform A and insulin-like growth factor-1 receptor in human acute myelogenous leukemia: effect of the dual-receptor inhibitor BMS-536924 in vitro. Cancer Res 2009, 69:7635-7643.
• [19]Dennis MD, Jefferson LS, Kimball SR: Role of p70S6K1-mediated phosphorylation of eIF4B and PDCD4 in the regulation of protein synthesis. J Biol Chem 2012, 287:42890-42899.
• [20]Tamburini J, Chapuis N, Bardet V, Park S, Sujobert P, Willems L, Ifrah N, Dreyfus F, Mayeux P, Lacombe C: Mammalian target of rapamycin (mTOR) inhibition activates phosphatidylinositol 3-kinase/Akt by up-regulating insulin-like growth factor-1 receptor signaling in acute myeloid leukemia: rationale for therapeutic inhibition of both pathways. Blood 2008, 111:379-382.
• [21]Pinton G, Manente AG, Angeli G, Mutti L, Moro L: Perifosine as a potential novel anti-cancer agent inhibits EGFR/MET-AKT axis in malignant pleural mesothelioma. PLoS One 2012, 7:e36856.
• [22]Li Z, Tan F, Liewehr DJ, Steinberg SM, Thiele CJ: In vitro and In vivo inhibition of Neuroblastoma tumor cell growth by AKT inhibitor Perifosine. J Natl Cancer Inst 2010, 102:758-770.
• [23]Bose P, Perkins EB, Honeycut C, Wellons MD, Stefan T, Jacobberger JW, Kontopodis E, Beumer JH, Egorin MJ, Imamura CK: Phase I trial of the combination of flavopiridol and imatinib mesylate in patients with Bcr-Abl + hematological malignancies. Cancer Chemother Pharmacol 2012, 69:1657-1667.
• [24]Vignot S, Faivre S, Aguirre D, Raymond E: mTOR-targeted therapy of cancer with rapamycin derivatives. Ann Oncol 2005, 16:525-537.
• [25]Tamburini J, Green AS, Bardet V, Chapuis N, Park S, Willems L, Uzunov M, Ifrah N, Dreyfus F, Lacombe C: Protein synthesis is resistant to rapamycin and constitutes a promising therapeutic target in acute myeloid leukemia. Blood 2009, 114:1618-1627.
• [26]Xu Q, Thompson JE, Carroll M: mTOR regulates cell survival after etoposide treatment in primary AML cells. Blood 2005, 106:4261-4268.
• [27]Altman JK, Sassano A, Kaur S, Glaser H, Kroczynska B, Redig AJ, Russo S, Barr S, Platanias LC: Dual mTORC2/mTORC1 targeting results in potent suppressive effects on acute myeloid leukemia (AML) progenitors. Clin Cancer Res 2011, 17:4378-4388.
• [28]Polak R, Buitenhuis M: The PI3K/PKB signaling module as key regulator of hematopoiesis: implications for therapeutic strategies in leukemia. Blood 2012, 119:911-923.
• [29]Guertin DA, Sabatini DM: The pharmacology of mTOR inhibition. Sci Signal 2009, 2(67):pe24. Apr 21
• [30]Janes MR, Limon JJ, So L, Chen J, Lim RJ, Chavez MA, Vu C, Lilly MB, Mallya S, Ong ST: Effective and selective targeting of leukemia cells using a TORC1/2 kinase inhibitor. Nat Med 2010, 16:205-213.
• [31]Willems L, Chapuis N, Puissant A, Maciel TT, Green AS, Jacque N, Vignon C, Park S, Guichard S, Herault O: The dual mTORC1 and mTORC2 inhibitor AZD8055 has anti-tumor activity in acute myeloid leukemia. Leukemia 2012, 26:1195-1202.
• [32]Sujobert P, Bardet V, Cornillet-Lefebvre P, Hayflick JS, Prie N, Verdier F, Vanhaesebroeck B, Muller O, Pesce F, Ifrah N: Essential role for the p110delta isoform in phosphoinositide 3-kinase activation and cell proliferation in acute myeloid leukemia. Blood 2005, 106:1063-1066.
• [33]Chapuis N, Tamburini J, Cornillet-Lefebvre P, Gillot L, Bardet V, Willems L, Park S, Green AS, Ifrah N, Dreyfus F: Autocrine IGF-1/IGF-1R signaling is responsible for constitutive PI3K/Akt activation in acute myeloid leukemia: therapeutic value of neutralizing anti-IGF-1R antibody. Haematologica 2010, 95:415-423.
• [34]Ma Z, Zhu L, Luo X, Zhai S, Li P, Wang X: Perifosine enhances mTORC1-targeted cancer therapy by activation of GSK3β in NSCLC cells. Cancer Biol Ther 2012, 13:1009-1017.
• [35]Tong Y, Liu YY, You LS, Qian WB: Perifosine induces protective autophagy and upregulation of ATG5 in human chronic myelogenous leukemia cells in vitro. Acta Pharmacol Sin 2012, 33:542-550.
• [36]Papa V, Tazzari PL, Chiarini F, Cappellini A, Ricci F, Billi AM, Evangelisti C, Ottaviani E, Martinelli G, Testoni N: Proapoptotic activity and chemosensitizing effect of the novel Akt inhibitor perifosine in acute myelogenous leukemia cells. Leukemia 2008, 22:147-160.
• [37]Pal SK, Reckamp K, Yu H, Figlin RA: Akt inhibitors in clinical development for the treatment of cancer. Expert Opin Investig Drugs 2010, 19:1355-1366.
• [38]Jin J, Liu H, Yang C, Li G, Liu X, Qian Q, Qian W: Effective gene-viral therapy of leukemia by a new fiber chimeric oncolytic adenovirus expressing TRAIL: in vitro and in vivo evaluation. Mol Cancer Ther 2009, 8:1387-1397.