Myeloma

Phospho-flow detection of constitutive and cytokine-induced pSTAT3/5, pAKT and pERK expression highlights novel prognostic biomarkers for patients with multiple myeloma

Article metrics

Subjects

Abstract

Identifying check points in cell signal transduction pathways has led to the development of new cancer therapies; however, relatively few studies have determined the diagnostic and prognostic significance of analysing phosphorylated signaling proteins in patient blood and bone marrow (BM) samples. This is the first comprehensive phospho-flow study of both constitutive and cytokine-induced pSTAT3, pSTAT5, pAKT and phosphorylated extracellular signal-regulated kinase (pERK) expression in malignant plasma cells of patients with monoclonal gammopathies. In diagnostic BM samples from 65 patients with multiple myeloma (MM), interleukin (IL)-6-induced pSTAT3 proved to be a new and independent prognostic biomarker for improved survival. When combined with the International Staging System, 6 subgroups demonstrated stratified median survivals from 9 to 72 months (χ2=34.3; P<0.0001). In contrast, constitutive expression of pSTAT3, pSTAT5, pAKT and pERK did not assist the differential diagnosis nor determine prognosis. High pSTAT3 expression was dependent on existing CD45 expression and pSTAT5 appeared to regulate IgG production. Phospho-flow cytometry could be used to screen for personalized therapy, although the lack of clinical significance of constitutive pSTAT3 levels suggests that pSTAT3 blockade may not be clinically relevant in MM. This study has revealed novel prognostic biomarkers and insights into the biology of signaling pathways in patients with MM.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1
Figure 2
Figure 3

References

  1. 1

    Darnell JE . STATs and gene regulation. Science 1997; 277: 1630–1635.

  2. 2

    Calò V, Migliavacca M, Bazan V, Macaluso M, Buscemi M, Gebbia N et al. STAT proteins: From normal control of cellular events to tumorigenesis. J Cell Physiol 2003; 197: 157–168.

  3. 3

    Sansone P, Bromberg J . Targeting interleukin-6/Jak/stat pathway in human malignancies. J Clin Oncol 2013; 30: 1005–1014.

  4. 4

    O’Shea JJ, Holland SM, Staudt L . Jaks and STATs in immunity, immunodeficiency and cancer. N Engl J Med 2013; 368: 161–170.

  5. 5

    Yu H, Pardoll D, Jove R . STATs in cancer inflammation and immunity: a leading role for STAT3. Nat Rev Cancer 2009; 9: 798–809.

  6. 6

    Vivanco I, Swayers CL . The phosphatidylinositol 3-kinase AKT pathway in human cancer. Nat Rev Cancer 2002; 2: 389–501.

  7. 7

    Downward J . Targeting RAS signalling pathways in cancer therapy. Nat Rev Cancer 2003; 3: 11–22.

  8. 8

    Fabbro D, Cowan-Jacob SW, Möbitz H, Martiny-Baron G . Targeting cancer with small-molecular-weight kinase inhibitors. Methods Mol Biol 2012; 795: 1–34.

  9. 9

    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.

  10. 10

    Leow CC-Y, Gerondakis S, Spencer A . MEK inhibitors as a chemotherapeutic intervention in multiple myeloma. Blood Cancer J 2013; 3: e105.

  11. 11

    Yang C, Lee H, Jove V, Deng J, Zhang W, Liu X et al. Prognostic significance of B-cells and pSTAT3 in patients with ovarian cancer. PLoS One 2013; 8: e54029.

  12. 12

    Kusaba T, Nakayama T, Yamazumi K, Yakata Y, Yoshizaki A, Inoue K et al. Activation of STAT3 is a marker of poor prognosis in human colorectal cancer. Oncol Rep 2006; 15: 1445–1451.

  13. 13

    Denley SM, Jamieson NB, McCall P, Oien KA, Morton JP, Carter CR et al. Activation of the IL-6R/Jak/stat pathway is associated with a poor outcome in resected pancreatic ductal adenocarcinoma. J Gastrointest Surg 2013; 17: 887–898.

  14. 14

    Leu CM, Wong FH, Chang C, Huang SF, Hu CP . Interleukin-6 acts as an antiapoptotic factor in human esophageal carcinoma cells through the activation of both STAT3 and mitogen-activated protein kinase pathways. Oncogene 2003; 22: 7809–7818.

  15. 15

    Xiong H, Du W, Wang JL, Hong J, Su WY, Tang JT et al. Constitutive activation of STAT3 is predictive of poor prognosis in human gastric cancer. J Mol Med (Berl) 2012; 90: 1037–1046.

  16. 16

    Woo S, Lee BL, Yoon J, Cho SJ, Baik TK, Chang MS et al. Constitutive activation of signal transducers and activators of transcription 3 correlates with better prognosis, cell proliferation and hypoxia-inducible factor-1α in human gastric cancer. Pathobiology 2011; 78: 295–301.

  17. 17

    Benekli M, Xia Z, Donohue KA, Ford LA, Pixley LA, Baer MR et al. Constitutive activity of signal transducer and activator of transcription 3 protein in acute myeloid leukemia blasts is associated with short disease-free survival. Blood 2002; 99: 252–257.

  18. 18

    Redell MS, Ruiz MJ, Gerbing RB, Alonzo TA, Lange BJ, Tweardy DJ et al. FACS analysis of Stat3/5 signaling reveals sensitivity to G-CSF and IL-6 as a significant prognostic factor in pediatric AML. Blood 2013; 121: 1083–1093.

  19. 19

    Hasegawa D, Bugarin C, Giordan M, Bresolin S, Longoni D, Micalizzi C et al. Validation of flow cytometric phospho-STAT5 as a diagnostic tool for juvenile myelomonocytic leukemia. Blood Cancer J 2013; 3: e160.

  20. 20

    Padron E, Painter JS, Kunigal S, Mailloux AW, McGraw K, McDaniel JM et al. GM-CSF–dependent pSTAT5 sensitivity is a feature with therapeutic potential in chronic myelomonocytic leukemia. Blood 2013; 121: 5068–5077.

  21. 21

    Stewart DA, Bahlis N, Mansoor A . pY-STAT3 and p53 expression predict outcome for poor prognosis diffuse large B-cell lymphoma treated with high dose chemotherapy and autologous stem cell transplantation. Leuk Lymphoma 2009; 50: 1276–1282.

  22. 22

    Wu ZL, Song YQ, Shi YF, Zhu J High nuclear expression of STAT3 is associated with unfavorable prognosis in diffuse large B-cell lymphoma. J Hematol Oncol 2011; 4: 31.

  23. 23

    Gupta M, Maurer MJ, Wellik LE, Law ME, Han JJ, Ozsan N et al. Expression of Myc, but not pSTAT3, is an adverse prognostic factor for diffuse large B-cell lymphoma treated with epratuzumab/R-CHOP. Blood 2012; 120: 4400–4406.

  24. 24

    Soldini D, Montagna C, Schüffler P, Martin V, Georgis A, Thiesler T et al. A new diagnostic algorithm for Burkitt and diffuse large B-cell lymphomas based on the expression of CSE1L and STAT3 and on MYC rearrangement predicts outcome. Ann Oncol 2013; 24: 193–201.

  25. 25

    Hodge LS, Ziesmer SC, Yang A-A, Secreto FJ, Novak AJ, Ansell S . Constitutive expression of STAT5A and STAT5B regulates IgM secretion in Waldenstrom’s macroglobulinemia. Blood 2014; 123: 1055–1058.

  26. 26

    Catlett-Falcone R, Landowski TH, Oshiro MM, Turkson J, Levitzki A, Savino R et al. Constitutive activation of Stat3 signaling confers resistance to apoptosis in human U266 myeloma cells. Immunity 1999; 10: 105–115.

  27. 27

    Quintanilla-Martinez L, Kremer M, Specht K, Calzada-Wack J, Nathrath N, Schaich R et al. Analysis of signal transducer and activator of transcription 3 (Stat 3) pathway in multiple myeloma. Am J Pathol 2003; 162: 1449–1461.

  28. 28

    Bharti AC, Shishodia S, Reuben JM, Weber D, Alexanian R, Raj-Vadhan S et al. Nuclear factor-kappaB and STAT3 are constitutively active in CD138+ cells derived from multiple myeloma patients, and suppression of these transcription factors leads to apoptosis. Blood 2004; 103: 3175–3184.

  29. 29

    Zollinger A, Stuhmer T, Chatterjee M, Gattenlöhner S, Haralambieva E, Muller-Hermelink HK et al. Combined functional and molecular analysis of tumor cell signaling defines 2 distinct myeloma subgroups: Akt-dependent and Akt-independent multiple myeloma. Blood 2008; 112: 3403–3411.

  30. 30

    Richardson PG, Wolf J, Jakubowiak A, Zonder J, Lonial S, Irwin D et al. Perifosine plus bortezomib and dexamethasone in patients with relapsed/refractory multiple myeloma previously treated with bortezomib: results of a multicenter phase I/II trial. J Clin Oncol 2011; 29: 44243–44249.

  31. 31

    Kumar S, Rajkumar SV, Greipp PR, Witzig T . CD45 expression by bone marrow cells in multiple myeloma: clinical and biological considerations. Leukemia 2005; 19: 1466–1470.

  32. 32

    Zhou Q, Yao Y, Ericson SG . The protein tyrosine phosphatase CD45 is required for interleukin 6 signaling in U266 myeloma cells. Int J Hematol 2004; 79: 63–73.

  33. 33

    Rawat R, Rainey GJ, Thompson CD, Frazier-Jessen MR, Brown RT, Nordan RP . Constitutive activation of STAT3 is associated with the acquisition of an interleukin 6-independent phenotype by murine plasmacytomas and hybridomas. Blood 2000; 96: 3514–3521.

  34. 34

    Brenne AT, Ro TB, Waage A, Sundan A, Borset M, Hjorth-Hansen H . Interleukin-21 is a growth and survival factor for human myeloma cells. Blood 2002; 99: 3756–3762.

  35. 35

    Iwamaru A, Szymanski S, Iwado E, Aoki H, Yokoyama T, Fokt I et al. A novel inhibitor of the STAT3 pathway induces apoptosis in malignant glioma cells both in vitro and in vivo. Oncogene 2007; 26: 2435–2444.

  36. 36

    Ferrajoli A, Faderl S, Van Q, Kantarjian HM, Talpaz M, Estrov Z et al. WP1066 disrupts Janus kinase-2 and induces caspase-dependent apoptosis in acute myelogenous leukemia cells. Cancer Res 2007; 67: 11291–11299.

  37. 37

    Abou-Ghazal M, Yang DS, Qiao W, Reina-Ortiz C, Wei J, Kong LY et al. The incidence, correlation with tumor-infiltrating inflammation, and prognosis of phosphorylated STAT3 expression in human gliomas. Clin Cancer Res 2008; 14: 8228–8235.

Download references

Acknowledgements

This work was funded by Cancer Institute of NSW and Sydney Foundation for Medical Research.

Author information

Correspondence to R Brown.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies this paper on the Leukemia website

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Further reading