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Tumor Biology

, Volume 37, Issue 9, pp 11947–11957 | Cite as

Dishevelled proteins are significantly upregulated in chronic lymphocytic leukaemia

  • Abdul Salam Khan
  • Mohammad Hojjat-Farsangi
  • Amir Hossein Daneshmanesh
  • Lotta Hansson
  • Parviz Kokhaei
  • Anders Österborg
  • Håkan Mellstedt
  • Ali Moshfegh
Original Article

Abstract

Dishevelled (DVL) proteins are components of the Wnt signalling pathways, and increased expression is associated with various malignancies. Information on DVLs in chronic lymphatic leukaemia (CLL) is limited. The aim of the present study was to investigate the role of DVLs in CLL cells and association with Wnt pathways downstream of ROR1. DVL1, 2 and 3 were exclusively expressed in CLL cells as compared to normal peripheral blood mononuclear cells (PBMCs). The expression of DVL1 and DVL3 proteins was significantly more pronounced in progressive than in non-progressive disease (p < 0.01), whereas the level of DVL2 was significantly higher in non-progressive as compared to progressive disease (p < 0.001). Treatment of CLL cells with anti-ROR1 specific monoclonal antibodies induced dephosphorylation of ROR1 as well as of tyrosine and serine residues of both DVL2 and DVL3. However, gene silencing of DVLs in the CLL cell line (EHEB) did not induce detectable apoptosis. Non-progressive CLL patients had a different protein activity pattern with regard to Wnt signalling pathway proteins as GSK-3β, β-catenin and AKT as compared to progressive disease. The DVL2 protein may play a role in the activation of signalling pathways in CLL during early stages of the disease, while DVL1 and 3 may have a role in later phases of the leukaemia.

Keywords

CLL DVL ROR1 Wnt 

Notes

Acknowledgments

This study was supported by grants from CLL Global Research Foundation, the Cancer and Allergy Foundation (149351, 149746, 150288), the Swedish Research Council (K2013-64X-21464-04-3), the Swedish Cancer Society (CAN 2009/852), the Cancer Society in Stockholm (121332), the King Gustav Vth Jubilee Fund (124272) and the Stockholm County Council (20120051). The secretarial help from Leila Relander is highly appreciated.

Authors’ contributions

ASK and AM designed the study, performed experiments, interpreted data and wrote the manuscript; MHF performed experiments and read the manuscript; AHDM read the manuscript; HM, AÖ and LH provided clinical samples and all read the manuscript; and HM supervised the study.

Compliance with ethical standards

Approval by the regional ethics committee (www.epn.se) was obtained as well as oral and written informed consent from the donors in accordance with the Helsinki Declaration.

Conflicts of interest

None

References

  1. 1.
    Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ. Cancer statistics, 2009. CA Cancer J Clin. 2009;59:225–49.CrossRefPubMedGoogle Scholar
  2. 2.
    Hamblin TJ, Davis Z, Gardiner A, Oscier DG, Stevenson FK. Unmutated Ig V(H) genes are associated with a more aggressive form of chronic lymphocytic leukemia. Blood. 1999;94:1848–54.PubMedGoogle Scholar
  3. 3.
    Baskar S, Kwong KY, Hofer T, Levy JM, Kennedy MG, Lee E, et al. Unique cell surface expression of receptor tyrosine kinase ROR1 in human B-cell chronic lymphocytic leukemia. Clin Cancer Res. 2008;14:396–404.CrossRefPubMedGoogle Scholar
  4. 4.
    Daneshmanesh AH, Mikaelsson E, Jeddi-Tehrani M, Bayat AA, Ghods R, Ostadkarampour M, et al. Ror1, a cell surface receptor tyrosine kinase is expressed in chronic lymphocytic leukemia and may serve as a putative target for therapy. Int J Cancer. 2008;123:1190–5.CrossRefPubMedGoogle Scholar
  5. 5.
    Fukuda T, Chen L, Endo T, Tang L, Lu D, Castro JE, et al. Antisera induced by infusions of autologous Ad-CD154-leukemia B cells identify ROR1 as an oncofetal antigen and receptor for Wnt5a. Proc Natl Acad Sci U S A. 2008;105:3047–52.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Forrester WC, Dell M, Perens E, Garriga G. A C. elegans ROR receptor tyrosine kinase regulates cell motility and asymmetric cell division. Nature. 1999;400:881–5.CrossRefPubMedGoogle Scholar
  7. 7.
    Al-Shawi R, Ashton SV, Underwood C, Simons JP. Expression of the Ror1 and Ror2 receptor tyrosine kinase genes during mouse development. Dev Genes Evol. 2001;211:161–71.CrossRefPubMedGoogle Scholar
  8. 8.
    Green JL, Kuntz SG, Sternberg PW. Ror receptor tyrosine kinases: orphans no more. Trends Cell Biol. 2008;18:536–44.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Morioka K, Tanikawa C, Ochi K, Daigo Y, Katagiri T, Kawano H, et al. Orphan receptor tyrosine kinase ROR2 as a potential therapeutic target for osteosarcoma. Cancer Sci. 2009;100:1227–33.CrossRefPubMedGoogle Scholar
  10. 10.
    Shabani M, Asgarian-Omran H, Vossough P, Sharifian RA, Faranoush M, Ghragozlou S, et al. Expression profile of orphan receptor tyrosine kinase (ROR1) and Wilms’ tumor gene 1 (WT1) in different subsets of B-cell acute lymphoblastic leukemia. Leuk Lymphoma. 2008;49:1360–7.CrossRefPubMedGoogle Scholar
  11. 11.
    Hudecek M, Schmitt TM, Baskar S, Lupo-Stanghellini MT, Nishida T, Yamamoto TN, et al. The B-cell tumor-associated antigen ROR1 can be targeted with T cells modified to express a ROR1-specific chimeric antigen receptor. Blood. 2010;116:4532–41.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Sato A, Yamamoto H, Sakane H, Koyama H, Kikuchi A. Wnt5a regulates distinct signalling pathways by binding to Frizzled2. EMBO J. 2010;29:41–54.CrossRefPubMedGoogle Scholar
  13. 13.
    Clevers H. Wnt/beta-catenin signaling in development and disease. Cell. 2006;127:469–80.CrossRefPubMedGoogle Scholar
  14. 14.
    Tian J, He H, Lei G. Wnt/beta-catenin pathway in bone cancers. Tumour Biol. 2014;35:9439–45.CrossRefPubMedGoogle Scholar
  15. 15.
    Bernassola F, Karin M, Ciechanover A, Melino G. The HECT family of E3 ubiquitin ligases: multiple players in cancer development. Cancer Cell. 2008;14:10–21.CrossRefPubMedGoogle Scholar
  16. 16.
    Bilic J, Huang YL, Davidson G, Zimmermann T, Cruciat CM, Bienz M, et al. Wnt induces LRP6 signalosomes and promotes dishevelled-dependent LRP6 phosphorylation. Science. 2007;316:1619–22.CrossRefPubMedGoogle Scholar
  17. 17.
    Moon RT. Wnt/beta-catenin pathway. Sci STKE 2005;2005:cm1.Google Scholar
  18. 18.
    Bryja V, Schulte G, Rawal N, Grahn A, Arenas E. Wnt-5a induces dishevelled phosphorylation and dopaminergic differentiation via a CK1-dependent mechanism. J Cell Sci. 2007;120:586–95.CrossRefPubMedGoogle Scholar
  19. 19.
    Klingensmith J, Yang Y, Axelrod JD, Beier DR, Perrimon N, Sussman DJ. Conservation of dishevelled structure and function between flies and mice: isolation and characterization of Dvl2. Mech Dev. 1996;58:15–26.CrossRefPubMedGoogle Scholar
  20. 20.
    Etheridge SL, Ray S, Li S, Hamblet NS, Lijam N, Tsang M, et al. Murine dishevelled 3 functions in redundant pathways with dishevelled 1 and 2 in normal cardiac outflow tract, cochlea, and neural tube development. PLoS Genet. 2008;4, e1000259.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Hamblet NS, Lijam N, Ruiz-Lozano P, Wang JB, Yang YS, Luo ZG, et al. Dishevelled 2 is essential for cardiac outflow tract development, somite segmentation and neural tube closure. Development. 2002;129:5827–38.CrossRefPubMedGoogle Scholar
  22. 22.
    Widelitz R. Wnt signaling through canonical and non-canonical pathways: recent progress. Growth Factors. 2005;23:111–6.CrossRefPubMedGoogle Scholar
  23. 23.
    Gao C, Chen YG. Dishevelled: the hub of Wnt signaling. Cell Signal. 2010;22:717–27.CrossRefPubMedGoogle Scholar
  24. 24.
    Jones C, Chen P. Planar cell polarity signaling in vertebrates. Bioessays. 2007;29:120–32.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Kuhl M. The Wnt/calcium pathway: biochemical mediators, tools and future requirements. Front Biosci. 2004;9:967–74.CrossRefPubMedGoogle Scholar
  26. 26.
    Gentile A, Lazzari L, Benvenuti S, Trusolino L, Comoglio PM. Ror1 is a pseudokinase that is crucial for met-driven tumorigenesis. Cancer Res. 2011;71:3132–41.CrossRefPubMedGoogle Scholar
  27. 27.
    Hallek M, Cheson BD, Catovsky D, Caligaris-Cappio F, Dighiero G, Dohner H, et al. International Workshop on Chronic Lymphocytic L: guidelines for the diagnosis and treatment of chronic lymphocytic leukemia: a report from the International Workshop on Chronic Lymphocytic Leukemia updating the National Cancer Institute-Working Group 1996 guidelines. Blood. 2008;111:5446–56.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Huang PY, Best OG, Almazi JG, Belov L, Davis ZA, Majid A, et al. Cell surface phenotype profiles distinguish stable and progressive chronic lymphocytic leukemia. Leuk Lymphoma. 2014;55:2085–92.CrossRefPubMedGoogle Scholar
  29. 29.
    Daneshmanesh AH, Farsangi MH, Moshfegh A, Khan S, Osterborg A, Mellstedt H. Apoptosis induction mediated through PI3-kinase/AKT/mTOR pathway using anti-ROR1 monoclonal antibody in chronic lymphocytic leukemia cells. ASCO Annual Meeting Abstracts. J Clin Oncol. 2013;31.Google Scholar
  30. 30.
    Hojjat-Farsangi M, Khan AS, Daneshmanesh AH, Moshfegh A, Sandin A, Mansouri L, et al. The tyrosine kinase receptor ROR1 is constitutively phosphorylated in chronic lymphocytic leukemia (CLL) cells. PLoS One. 2013;8, e78339.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Reya T, Clevers H. Wnt signalling in stem cells and cancer. Nature. 2005;434:843–50.CrossRefPubMedGoogle Scholar
  32. 32.
    Miller JR, Hocking AM, Brown JD, Moon RT. Mechanism and function of signal transduction by the Wnt/beta-catenin and Wnt/Ca2+ pathways. Oncogene. 1999;18:7860–72.CrossRefPubMedGoogle Scholar
  33. 33.
    Novak A, Dedhar S. Signaling through beta-catenin and Lef/Tcf. Cell Mol Life Sci. 1999;56:523–37.CrossRefPubMedGoogle Scholar
  34. 34.
    Grumolato L, Liu G, Mong P, Mudbhary R, Biswas R, Arroyave R, et al. Canonical and noncanonical Wnts use a common mechanism to activate completely unrelated coreceptors. Genes Dev. 2010;24:2517–30.CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Zhang A, He S, Sun X, Ding L, Bao X, Wang N. Wnt5a promotes migration of human osteosarcoma cells by triggering a phosphatidylinositol-3 kinase/Akt signals. Cancer Cell Int. 2014;14:15.CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Cuni S, Perez-Aciego P, Perez-Chacon G, Vargas JA, Sanchez A, Martin-Saavedra FM, et al. A sustained activation of PI3k/NF-kappaB pathway is critical for the survival of chronic lymphocytic leukemia B cells. Leukemia. 2004;18:1391–400.CrossRefPubMedGoogle Scholar
  37. 37.
    Barragan M, Bellosillo B, Campas C, Colomer D, Pons G, Gil J. Involvement of protein kinase C and phosphatidylinositol 3-kinase pathways in the survival of B-cell chronic lymphocytic leukemia cells. Blood. 2002;99:2969–76.CrossRefPubMedGoogle Scholar
  38. 38.
    Jones DT, Ganeshaguru K, Anderson RJ, Jackson TR, Bruckdorfer KR, Low SY, et al. Albumin activates the AKT signaling pathway and protects B-chronic lymphocytic leukemia cells from chlorambucil- and radiation-induced apoptosis. Blood. 2003;101:3174–80.CrossRefPubMedGoogle Scholar
  39. 39.
    Petlickovski A, Laurenti L, Li X, Marietti S, Chiusolo P, Sica S, et al. Sustained signaling through the B-cell receptor induces Mcl-1 and promotes survival of chronic lymphocytic leukemia B cells. Blood. 2005;105:4820–7.CrossRefPubMedGoogle Scholar
  40. 40.
    Kawasaki A, Torii K, Yamashita Y, Nishizawa K, Kanekura K, Katada M, et al. Wnt5a promotes adhesion of human dermal fibroblasts by triggering a phosphatidylinositol-3 kinase/Akt signal. Cell Signal. 2007;19:2498–506.CrossRefPubMedGoogle Scholar
  41. 41.
    Memarian A, Hojjat-Farsangi M, Asgarian-Omran H, Younesi V, Jeddi-Tehrani M, Sharifian RA, et al. Variation in WNT genes expression in different subtypes of chronic lymphocytic leukemia. Leuk Lymphoma. 2009;50:2061–70.CrossRefPubMedGoogle Scholar
  42. 42.
    Simons M, Mlodzik M. Planar cell polarity signaling: from fly development to human disease. Annu Rev Genet. 2008;42:517–40.CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Thomas C, Strutt D. The roles of the cadherins fat and dachsous in planar polarity specification in Drosophila. Dev Dyn. 2012;241:27–39.CrossRefPubMedGoogle Scholar
  44. 44.
    Christofori G. New signals from the invasive front. Nature. 2006;441:444–50.CrossRefPubMedGoogle Scholar
  45. 45.
    Zhao Y, Yang ZQ, Wang Y, Miao Y, Liu Y, Dai SD, et al. Dishevelled-1 and dishevelled-3 affect cell invasion mainly through canonical and noncanonical Wnt pathway, respectively, and associate with poor prognosis in nonsmall cell lung cancer. Mol Carcinog. 2010;49:760–70.CrossRefPubMedGoogle Scholar
  46. 46.
    Gutierrez Jr A, Tschumper RC, Wu X, Shanafelt TD, Eckel-Passow J, Huddleston 3rd PM, et al. Lef-1 is a prosurvival factor in chronic lymphocytic leukemia and is expressed in the preleukemic state of monoclonal B-cell lymphocytosis. Blood. 2010;116:2975–83.CrossRefPubMedPubMedCentralGoogle Scholar
  47. 47.
    He TY, Wu DW, Lin PL, Wang L, Huang CC, Chou MC, et al. Ddx3 promotes tumor invasion in colorectal cancer via the ck1epsilon/Dvl2 axis. Sci Rep. 2016;6:21483.CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Kaucka M, Plevova K, Pavlova S, Janovska P, Mishra A, Verner J, et al. The planar cell polarity pathway drives pathogenesis of chronic lymphocytic leukemia by the regulation of B-lymphocyte migration. Cancer Res. 2013;73:1491–501.CrossRefPubMedGoogle Scholar
  49. 49.
    Katoh M. WNT/PCP signaling pathway and human cancer (review). Oncol Rep. 2005;14:1583–8.PubMedGoogle Scholar
  50. 50.
    Mittal AK, Chaturvedi NK, Rai KJ, Gilling-Cutucache CE, Nordgren TM, Moragues M, et al. Chronic lymphocytic leukemia cells in a lymph node microenvironment depict molecular signature associated with an aggressive disease. Mol Med. 2014;20:290–301.CrossRefPubMedPubMedCentralGoogle Scholar
  51. 51.
    Wynshaw-Boris A. Dishevelled: in vivo roles of a multifunctional gene family during development. Curr Top Dev Biol. 2012;101:213–35.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  • Abdul Salam Khan
    • 1
  • Mohammad Hojjat-Farsangi
    • 1
  • Amir Hossein Daneshmanesh
    • 1
  • Lotta Hansson
    • 1
    • 2
  • Parviz Kokhaei
    • 1
    • 3
  • Anders Österborg
    • 1
    • 2
  • Håkan Mellstedt
    • 1
    • 4
  • Ali Moshfegh
    • 1
  1. 1.Department of Oncology-Pathology, Immune and Gene Therapy Lab, Cancer Center Karolinska (CCK)Karolinska University Hospital Solna and Karolinska InstitutetStockholmSweden
  2. 2.Department of HematologyKarolinska University Hospital SolnaStockholmSweden
  3. 3.Cancer Research Center and Department of ImmunologySemnan University of Medical SciencesSemnanIran
  4. 4.Cancer Centre Karolinska, Department of OncologyKarolinska University Hospital SolnaStockholmSweden

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