, Volume 19, Issue 1, pp 201–209 | Cite as

The putative BH3 mimetic S1 sensitizes leukemia to ABT-737 by increasing reactive oxygen species, inducing endoplasmic reticulum stress, and upregulating the BH3-only protein NOXA

  • Ryan Soderquist
  • Alexandre A. Pletnev
  • Alexey V. Danilov
  • Alan Eastman
Original Paper


S1 is a putative BH3 mimetic proposed to inhibit BCL2 and MCL1 based on cell-free assays. However, we previously demonstrated that it failed to inhibit BCL2 or induce apoptosis in chronic lymphocytic leukemia (CLL) cells, which are dependent on BCL2 for survival. In contrast, we show here that S1 rapidly increases reactive oxygen species, initiates endoplasmic reticulum stress, and upregulates the BH3-only protein NOXA. The BCL2 inhibitors, ABT-737, ABT-263, and ABT-199, have demonstrated pro-apoptotic efficacy in cell lines, while ABT-263 and ABT-199 have demonstrated efficacy in early clinical trials. Resistance to these inhibitors arises from the upregulation of anti-apoptotic factors, such as MCL1, BFL1, and BCLXL. This resistance can be induced by co-culturing CLL cells on a stromal cell line that mimics the microenvironment found in patients. Since NOXA can inhibit MCL1, BFL1, and BCLXL, we hypothesized that S1 may overcome resistance to ABT-737. Here we demonstrate that S1 induces NOXA-dependent sensitization to ABT-737 in a human promyelocytic leukemia cell line (NB4). Furthermore, S1 sensitized CLL cells to ABT-737 ex vivo, and overcame resistance to ABT-737 induced by co-culturing CLL cells with stroma.


BCL2 MCL1 BCLXL NOXA Reactive oxygen species ATF3 



This research was supported by a Translational Research Grant from the Leukemia and Lymphoma Society and a Cancer Center Support Grant to the Norris Cotton Cancer Center (NIH CA23108). Support to A.V.D. was provided by a National Cancer Institute new faculty award (NIH CA023108-31S4) to the Norris Cotton Cancer Center.

Conflict of interest


Supplementary material

10495_2013_910_MOESM1_ESM.pdf (724 kb)
Supplementary material 1 (PDF 723 kb)


  1. 1.
    Hanahan D, Weinberg Robert A (2011) Hallmarks of cancer: the next generation. Cell 144:646–674PubMedCrossRefGoogle Scholar
  2. 2.
    Dewson G, Kluck RM (2009) Mechanisms by which Bak and Bax permeabilise mitochondria during apoptosis. J Cell Sci 122:2801–2808PubMedCrossRefGoogle Scholar
  3. 3.
    Walensky LD (2006) BCL-2 in the crosshairs: tipping the balance of life and death. Cell Death Differ 13:1339–1350PubMedCrossRefGoogle Scholar
  4. 4.
    Lomonosova E, Chinnadurai G (2008) BH3-only proteins in apoptosis and beyond: an overview. Oncogene 27:S2–S19PubMedCentralPubMedCrossRefGoogle Scholar
  5. 5.
    Roberts AW, Seymour JF, Brown JR, Wierda WG, Kipps TJ, Khaw SL, Carney DA, He SZ, Huang DCS, Xiong H, Cui Y, Busman TA, McKeegan EM, Krivoshik AP, Enschede SH, Humerickhouse R (2012) Substantial susceptibility of chronic lymphocytic leukemia to BCL2 inhibition: results of a phase I study of Navitoclax in patients with relapsed or refractory disease. J Clin Oncol 30:488–496PubMedCrossRefGoogle Scholar
  6. 6.
    Gandhi L, Camidge DR, Ribeiro de Oliveira M, Bonomi P, Gandara D, Khaira D, Hann CL, McKeegan EM, Litvinovich E, Hemken PM, Dive C, Enschede SH, Nolan C, Chiu Y-L, Busman T, Xiong H, Krivoshik AP, Humerickhouse R, Shapiro GI, Rudin CM (2011) Phase I study of Navitoclax (ABT-263), a novel Bcl-2 family inhibitor, in patients with small-cell lung cancer and other solid tumors. J Clin Oncol 29:909–916PubMedCrossRefGoogle Scholar
  7. 7.
    Yecies D, Carlson NE, Deng J, Letai A (2010) Acquired resistance to ABT-737 in lymphoma cells that up-regulate MCL-1 and BFL-1. Blood 115:3304–3313PubMedCrossRefGoogle Scholar
  8. 8.
    Albershardt TC, Salerni BL, Soderquist RS, Bates DJP, Pletnev AA, Kisselev AF, Eastman A (2011) Multiple BH3 mimetics antagonize antiapoptotic MCL1 protein by inducing the endoplasmic reticulum stress response and up-regulating BH3-only protein NOXA. J Biol Chem 286:24882–24895PubMedCrossRefGoogle Scholar
  9. 9.
    van Delft MF, Wei AH, Mason KD, Vandenberg CJ, Chen L, Czabotar PE, Willis SN, Scott CL, Day CL, Cory S, Adams JM, Roberts AW, Huang DC (2006) The BH3 mimetic ABT-737 targets selective Bcl-2 proteins and efficiently induces apoptosis via Bak/Bax if Mcl-1 is neutralized. Cancer Cell 10:389–399PubMedCentralPubMedCrossRefGoogle Scholar
  10. 10.
    Zhang Z, Jin L, Qian X, Wei M, Wang Y, Wang J, Yang Y, Xu Q, Xu Y, Liu F (2007) Novel Bcl-2 inhibitors: discovery and mechanism study of small organic apoptosis-inducing agents. Chem Bio Chem 8:113–121PubMedCrossRefGoogle Scholar
  11. 11.
    Song T, Chang X, Zhang Z, Liu Y, Shen X (2012) S1, a novel pan-BH3 mimetic, induces apoptosis in Mcl-1-overexpressing cells through Bak. J Pharmacol Sci 119:330–340PubMedCrossRefGoogle Scholar
  12. 12.
    Salerni BL, Bates DJ, Albershardt TC, Lowrey CH, Eastman A (2010) Vinblastine induces acute, cell cycle phase-independent apoptosis in some leukemias and lymphomas and can induce acute apoptosis in others when Mcl-1 is suppressed. Mol Cancer Ther 9:791–802PubMedCentralPubMedCrossRefGoogle Scholar
  13. 13.
    Bates DJP, Salerni BL, Lowrey CH, Eastman A (2011) Vinblastine sensitizes leukemia cells to cyclin-dependent kinase inhibitors, inducing acute cell cycle phase-independent apoptosis. Cancer Biol Ther 12:314–325PubMedCrossRefGoogle Scholar
  14. 14.
    Neron S, Pelletier A, Chevrier M-C, Monier G, Lemieux R, Darveau A (1996) Induction of LFA-1 independent human B cell proliferation and differentiation by binding of CD40 with its ligand. Immunol Invest 25:79–89PubMedCrossRefGoogle Scholar
  15. 15.
    Lee EF, Czabotar PE, van Delft MF, Michalak EM, Boyle MJ, Willis SN, Puthalakath H, Bouillet P, Colman PM, Huang DCS, Fairlie WD (2008) A novel BH3 ligand that selectively targets Mcl-1 reveals that apoptosis can proceed without Mcl-1 degradation. J Cell Biol 180:341–355PubMedCrossRefGoogle Scholar
  16. 16.
    Yoshida H, Matsui T, Yamamoto A, Okada T, Mori K (2001) XBP1 mRNA is induced by ATF6 and spliced by IRE1 in response to ER stress to produce a highly active transcription factor. Cell 107:881–891PubMedCrossRefGoogle Scholar
  17. 17.
    Wang Q, Mora-Jensen H, Weniger MA, Perez-Galan P, Wolford C, Hai T, Ron D, Chen W, Trenkle W, Wiestner A, Ye Y (2009) ERAD inhibitors integrate ER stress with an epigenetic mechanism to activate BH3-only protein NOXA in cancer cells. Proc Natl Acad Sci 106:2200–2205PubMedCrossRefGoogle Scholar
  18. 18.
    Ding W, Yang L, Zhang M, Gu Y (2012) Reactive oxygen species-mediated endoplasmic reticulum stress contributes to aldosterone-induced apoptosis in tubular epithelial cells. Biochem Biophys Res Commun 418:451–456PubMedCrossRefGoogle Scholar
  19. 19.
    Zafarullah M, Li WQ, Sylvester J, Ahmad M (2003) Molecular mechanisms of N-acetylcysteine actions. Cell Mol Life Sci 60:6–20PubMedCrossRefGoogle Scholar
  20. 20.
    Mason KD, Khaw SL, Rayeroux KC, Chew E, Lee EF, Fairlie WD, Grigg AP, Seymour JF, Szer J, Huang DCS, Roberts AW (2009) The BH3 mimetic compound, ABT-737, synergizes with a range of cytotoxic chemotherapy agents in chronic lymphocytic leukemia. Leukemia 23:2034–2041PubMedCrossRefGoogle Scholar
  21. 21.
    Vogler M, Butterworth M, Majid A, Walewska RJ, Sun X-M, Dyer MJS, Cohen GM (2009) Concurrent up-regulation of BCL-XL and BCL2A1 induces approximately 1000-fold resistance to ABT-737 in chronic lymphocytic leukemia. Blood 113:4403–4413PubMedCrossRefGoogle Scholar
  22. 22.
    Vogler M, Weber K, Dinsdale D, Schmitz I, Schulze-Osthoff K, Dyer MJS, Cohen GM (2009) Different forms of cell death induced by putative BCL2 inhibitors. Cell Death Differ 16:1030–1039PubMedCrossRefGoogle Scholar
  23. 23.
    Stewart ML, Fire E, Keating AE, Walensky LD (2010) The MCL-1 BH3 helix is an exclusive MCL-1 inhibitor and apoptosis sensitizer. Nat Chem Biol 6:595–601PubMedCentralPubMedCrossRefGoogle Scholar
  24. 24.
    Doi K, Li R, Sung S–S, Wu H, Liu Y, Manieri W, Krishnegowda G, Awwad A, Dewey A, Liu X, Amin S, Cheng C, Qin Y, Schonbrunn E, Daughdrill G, Loughran TP, Sebti S, Wang H-G (2012) Discovery of Marinopyrrole A (Maritoclax) as a selective Mcl-1 antagonist that overcomes ABT-737 resistance by binding to and targeting Mcl-1 for proteasomal degradation. J Biol Chem 287:10224–10235PubMedCrossRefGoogle Scholar
  25. 25.
    Tromp JM, Geest CR, Breij ECW, Elias JA, van Laar J, Luijks DM, Kater AP, Beaumont T, van Oers MHJ, Eldering E (2012) Tipping the Noxa/Mcl-1 balance overcomes ABT-737 resistance in chronic lymphocytic leukemia. Clin Cancer Res 18:487–498PubMedCrossRefGoogle Scholar
  26. 26.
    Lucas KM, Mohana-Kumaran N, Lau D, Zhang XD, Hersey P, Huang DC, Weninger W, Haass NK, Allen JD (2012) Modulation of NOXA and MCL-1 as a strategy for sensitizing melanoma cells to the BH3-mimetic ABT-737. Clin Cancer Res 18:783–795PubMedCrossRefGoogle Scholar
  27. 27.
    Cairns RA, Harris IS, Mak TW (2011) Regulation of cancer cell metabolism. Nat Rev Cancer 11:85–95PubMedCrossRefGoogle Scholar
  28. 28.
    Raj L, Ide T, Gurkar AU, Foley M, Schenone M, Li X, Tolliday NJ, Golub TR, Carr SA, Shamji AF, Stern AM, Mandinova A, Schreiber SL, Lee SW (2011) Selective killing of cancer cells by a small molecule targeting the stress response to ROS. Nature 475:231–234PubMedCentralPubMedCrossRefGoogle Scholar
  29. 29.
    Trachootham D, Alexandre J, Huang P (2009) Targeting cancer cells by ROS-mediated mechanisms: a radical therapeutic approach? Nat Rev Drug Discov 8:579–591PubMedCrossRefGoogle Scholar
  30. 30.
    Song JH, Kandasamy K, Zemskova M, Lin Y-W, Kraft AS (2011) The BH3 mimetic ABT-737 induces cancer cell senescence. Cancer Res 71:506–515PubMedCentralPubMedCrossRefGoogle Scholar
  31. 31.
    Cruickshanks N, Tang Y, Booth L, Hamed H, Grant S, Dent P (2012) Lapatinib and Obatoclax kill breast cancer cells through reactive oxygen species-dependent endoplasmic reticulum stress. Mol Pharmacol 82:1217–1229PubMedCrossRefGoogle Scholar
  32. 32.
    Zhang Z, Song T, Zhang T, Gao J, Wu G, An L, Du G (2011) A novel BH3 mimetic S1 potently induces Bax/Bak-dependent apoptosis by targeting both Bcl-2 and Mcl-1. Int J Cancer 128:1724–1735PubMedCrossRefGoogle Scholar
  33. 33.
    Zhong JT, Xu Y, Yi HW, Su J, Yu HM, Xiang XY, Li XN, Zhang ZC, Sun LK (2012) The BH3 mimetic S1 induces autophagy through ER stress and disruption of Bcl-2/Beclin 1 interaction in human glioma U251 cells. Cancer Lett 323:180–187PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Ryan Soderquist
    • 1
  • Alexandre A. Pletnev
    • 2
  • Alexey V. Danilov
    • 3
  • Alan Eastman
    • 1
  1. 1.Department of Pharmacology and ToxicologyNorris Cotton Cancer Center, Geisel School of Medicine at DartmouthLebanonUSA
  2. 2.Department of ChemistryDartmouth CollegeHanoverUSA
  3. 3.Department of MedicineNorris Cotton Cancer Center, Geisel School of Medicine at DartmouthLebanonUSA

Personalised recommendations