Low expression of pro-apoptotic Bcl-2 family proteins sets the apoptotic threshold in Waldenström macroglobulinemia

Article metrics


Waldenström macroglobulinemia (WM) is a proliferative disorder of IgM-secreting, lymphoplasmacytoid cells that inhabit the lymph nodes and bone marrow. The disease carries a high prevalence of activating mutations in MyD88 (91%) and CXCR4 (28%). Because signaling through these pathways leads to Bcl-xL induction, we examined Bcl-2 family expression in WM patients and cell lines. Unlike other B-lymphocyte-derived malignancies, which become dependent on expression of anti-apoptotic proteins to counter expression of pro-apoptotic proteins, WM samples expressed both pro- and anti-apoptotic Bcl-2 proteins at low levels similar to their normal B-cell and plasma cell counterparts. Three WM cell lines expressed pro-apoptotic Bcl-2 family members Bim or Bax and Bak at low levels, which determined their sensitivity to inducers of intrinsic apoptosis. In two cell lines, miR-155 upregulation, which is common in WM, was responsible for the inhibition of FOXO3a and Bim expression. Both antagonizing miR-155 to induce Bim and proteasome inhibition increased the sensitivity to ABT-737 in these lines indicating a lowering of the apoptotic threshold. In this manner, treatments that increase pro-apoptotic protein expression increase the efficacy of agents treated in combination in addition to direct killing.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7


  1. 1

    Chipuk JE, Moldoveanu T, Llambi F, Parsons MJ, Green DR . The BCL-2 family reunion. Mol Cell 2010; 37: 299–310.

  2. 2

    Ren D, Tu HC, Kim H, Wang GX, Bean GR, Takeuchi O et al. BID, BIM, and PUMA are essential for activation of the BAX- and BAK-dependent cell death program. Science 2010; 330: 1390–1393.

  3. 3

    Wei MC, Zong WX, Cheng EH, Lindsten T, Panoutsakopoulou V, Ross AJ et al. Proapoptotic BAX and BAK: a requisite gateway to mitochondrial dysfunction and death. Science 2001; 292: 727–730.

  4. 4

    Danial NN, Korsmeyer SJ . Cell death: critical control points. Cell 2004; 116: 205–219.

  5. 5

    Certo M, Del Gaizo Moore V, Nishino M, Wei G, Korsmeyer S, Armstrong SA et al. Mitochondria primed by death signals determine cellular addiction to antiapoptotic BCL-2 family members. Cancer Cell 2006; 9: 351–365.

  6. 6

    Strasser A, Harris AW, Bath ML, Cory S . Novel primitive lymphoid tumours induced in transgenic mice by cooperation between myc and bcl-2. Nature 1990; 348: 331–333.

  7. 7

    Beverly LJ, Varmus HE . MYC-induced myeloid leukemogenesis is accelerated by all six members of the antiapoptotic BCL family. Oncogene 2009; 28: 1274–1279.

  8. 8

    Ni Chonghaile T, Sarosiek KA, Vo TT, Ryan JA, Tammareddi A, Moore Vdel G et al. Pretreatment mitochondrial priming correlates with clinical response to cytotoxic chemotherapy. Science 2011; 334: 1129–1133.

  9. 9

    Herrinton LJ, Weiss NS . Incidence of Waldenstrom's macroglobulinemia. Blood 1993; 82: 3148–3150.

  10. 10

    Ghobrial IM, Gertz MA, Fonseca R . Waldenstrom macroglobulinaemia. Lancet Oncol 2003; 4: 679–685.

  11. 11

    Treon SP, Xu L, Yang G, Zhou Y, Liu X, Cao Y et al. MYD88 L265P somatic mutation in Waldenstrom's macroglobulinemia. N Engl J Med 2012; 367: 826–833.

  12. 12

    Poulain S, Roumier C, Decambron A, Renneville A, Herbaux C, Bertrand E et al. MYD88 L265P mutation in Waldenstrom macroglobulinemia. Blood 2013; 121: 4504–4511.

  13. 13

    Gaudette BT, Iwakoshi NN, Boise LH . Bcl-xL protein protects from C/EBP homologous protein (CHOP)-dependent apoptosis during plasma cell differentiation. J Biol Chem 2014; 289: 23629–23640.

  14. 14

    Gutierrez NC, Ocio EM, de Las Rivas J, Maiso P, Delgado M, Ferminan E et al. Gene expression profiling of B lymphocytes and plasma cells from Waldenstrom's macroglobulinemia: comparison with expression patterns of the same cell counterparts from chronic lymphocytic leukemia, multiple myeloma and normal individuals. Leukemia 2007; 21: 541–549.

  15. 15

    Ditzel Santos D, Ho AW, Tournilhac O, Hatjiharissi E, Leleu X, Xu L et al. Establishment of BCWM.1 cell line for Waldenstrom's macroglobulinemia with productive in vivo engraftment in SCID-hu mice. Exp Hematol 2007; 35: 1366–1375.

  16. 16

    Hodge LS, Novak AJ, Grote DM, Braggio E, Ketterling RP, Manske MK et al. Establishment and characterization of a novel Waldenstrom macroglobulinemia cell line, MWCL-1. Blood 2011; 117: e190–e197.

  17. 17

    Chitta KS, Paulus A, Ailawadhi S, Foster BA, Moser MT, Starostik P et al. Development and characterization of a novel human Waldenstrom macroglobulinemia cell line: RPCI-WM1, Roswell Park Cancer Institute—Waldenstrom Macroglobulinemia 1. Leuk Lymphoma 2013; 54: 387–396.

  18. 18

    Morales AA, Gutman D, Lee KP, Boise LH . BH3-only proteins Noxa, Bmf, and Bim are necessary for arsenic trioxide-induced cell death in myeloma. Blood 2008; 111: 5152–5162.

  19. 19

    Oltersdorf T, Elmore SW, Shoemaker AR, Armstrong RC, Augeri DJ, Belli BA et al. An inhibitor of Bcl-2 family proteins induces regression of solid tumours. Nature 2005; 435: 677–681.

  20. 20

    Roccaro AM, Sacco A, Chen C, Runnels J, Leleu X, Azab F et al. microRNA expression in the biology, prognosis, and therapy of Waldenstrom macroglobulinemia. Blood 2009; 113: 4391–4402.

  21. 21

    Lewis BP, Burge CB, Bartel DP . Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 2005; 120: 15–20.

  22. 22

    Kong W, He L, Coppola M, Guo J, Esposito NN, Coppola D et al. MicroRNA-155 regulates cell survival, growth, and chemosensitivity by targeting FOXO3a in breast cancer. J Biol Chem 2010; 285: 17869–17879.

  23. 23

    Yamanaka Y, Tagawa H, Takahashi N, Watanabe A, Guo YM, Iwamoto K et al. Aberrant overexpression of microRNAs activate AKT signaling via down-regulation of tumor suppressors in natural killer-cell lymphoma/leukemia. Blood 2009; 114: 3265–3275.

  24. 24

    Zhang Y, Roccaro AM, Rombaoa C, Flores L, Obad S, Fernandes SM et al. LNA-mediated anti-miR-155 silencing in low-grade B-cell lymphomas. Blood 2012; 120: 1678–1686.

  25. 25

    Premkumar DR, Jane EP, DiDomenico JD, Vukmer NA, Agostino NR, Pollack IF . ABT-737 synergizes with bortezomib to induce apoptosis, mediated by Bid cleavage, Bax activation, and mitochondrial dysfunction in an Akt-dependent context in malignant human glioma cell lines. J Pharmacol Exp Ther 2012; 341: 859–872.

  26. 26

    Gertz MA . Waldenstrom macroglobulinemia: 2013 update on diagnosis, risk stratification, and management. Am J Hematol 2013; 88: 703–711.

  27. 27

    Treon SP, Tripsas CK, Meid K, Kanan S, Sheehy P, Chuma S et al. Carfilzomib, rituximab and dexamethasone (CaRD) is active and offers a neuropathy-sparing approach for proteasome-inhibitor based therapy in Waldenstrom's macroglobulinemia. Blood 2014; 124: 503–510.

  28. 28

    Roccaro AM, Sacco A, Aujay M, Ngo HT, Azab AK, Azab F et al. Selective inhibition of chymotrypsin-like activity of the immunoproteasome and constitutive proteasome in Waldenstrom macroglobulinemia. Blood 2010; 115: 4051–4060.

  29. 29

    Boise LH . Aiming at WM with both barrels blocked. Blood 2010; 115: 4007–4008.

  30. 30

    Morales AA, Kurtoglu M, Matulis SM, Liu J, Siefker D, Gutman DM et al. Distribution of Bim determines Mcl-1 dependence or codependence with Bcl-xL/Bcl-2 in Mcl-1-expressing myeloma cells. Blood 2011; 118: 1329–1339.

  31. 31

    Boise LH, Minn AJ, Noel PJ, June CH, Accavitti MA, Lindsten T et al. CD28 costimulation can promote T cell survival by enhancing the expression of Bcl-xL. Immunity 1995; 3: 87–98.

Download references


We thank Stephen Ansell and Kelvin Lee for cell lines and reagents. This work was supported by: R01 CA127910 and R01 CA129968 as well as funding from the TJ Martell Foundation (LHB), the Leukemia & Lymphoma Society (AAC), the International Waldenstrom Macroglobulinemia (AAC), and the Comité du Septentrion de la Ligue contre le Cancer (XL). LHB is a GRA Distinguished Cancer Scientist. ABT-737 and ABT-199 were a generous gift of Abbvie, (North Chicago, IL, USA).

Author information

Correspondence to L H Boise.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies this paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Further reading