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p66Shc deficiency enhances CXCR4 and CCR7 recycling in CLL B cells by facilitating their dephosphorylation-dependent release from β-arrestin at early endosomes

Oncogene volume 37, pages 1534–1550 (2018)Cite this article

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Abstract

Neoplastic cell traffic abnormalities are central to the pathogenesis of chronic lymphocytic leukemia (CLL). Enhanced CXC chemokine receptor-4 (CXCR4) and chemokine receptor-7 (CCR7) recycling contributes to the elevated surface levels of these receptors on CLL cells. Here we have addressed the role of p66Shc, a member of the Shc family of protein adaptors the expression of which is defective in CLL cells, in CXCR4/CCR7 recycling. p66Shc reconstitution in CLL cells reduced CXCR4/CCR7 recycling, lowering their surface levels and attenuating B-cell chemotaxis, due to their accumulation in Rab5+ endosomes as serine-phosphoproteins bound to β-arrestin. This results from the ability of p66Shc to inhibit Ca2+ and PP2B-dependent CXCR4/CCR7 dephosphorylation and β-arrestin release. We also show that ibrutinib, a Btk inhibitor that promotes leukemic cell mobilization from lymphoid organs, reverses the CXCR4/CCR7 recycling abnormalities in CLL cells by increasing p66Shc expression. These results, identifying p66Shc as a regulator of CXCR4/CCR7 recycling in B cells, underscore the relevance of its deficiency to CLL pathogenesis and provide new clues to the mechanisms underlying the therapeutic effects of ibrutinib.

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References

  1. Scarfo L, Ferreri AJ, Ghia P. Chronic lymphocytic leukaemia. Crit Rev Oncol Hematol. 2016;104:169–82.

    Article  Google Scholar 

  2. Burger JA, Gribben JG, The microenvironment in chronic lymphocytic leukemia (CLL) and other B cell malignancies: insight into disease biology and new targeted therapies. Semin Cancer Biol. 2014;24:71–81.

    Article  CAS  Google Scholar 

  3. Munk Pedersen I, Reed J. Microenvironmental interactions and survival of CLL B-cells. Leuk Lymphoma. 2004;45:2365–72.

    Article  CAS  Google Scholar 

  4. Kehrl JH, Hwang IY, Park C. Chemoattract receptor signaling and its role in lymphocyte motility and trafficking. Curr Top Microbiol Immunol. 2009;334:107–27.

    CAS  PubMed  Google Scholar 

  5. Kucia M, Jankowski K, Reca R, Wysoczynski M, Bandura L, Allendorf DJ, et al. CXCR4-SDF-1 signalling, locomotion, chemotaxis and adhesion. J Mol Histol. 2004;35:233–45.

    Article  CAS  Google Scholar 

  6. Comerford I, Harata-Lee Y, Bunting MD, Gregor C, Kara EE, McColl SR. A myriad of functions and complex regulation of the CCR7/CCL19/CCL21 chemokine axis in the adaptive immune system. Cytokine Growth Factor Rev. 2013;24:269–83.

    Article  CAS  Google Scholar 

  7. Kurtova AV, Balakrishnan K, Chen R, Ding W, Schnabl S, Quiroga MP, et al. Diverse marrow stromal cells protect CLL cells from spontaneous and drug-induced apoptosis: development of a reliable and reproducible system to assess stromal cell adhesion-mediated drug resistance. Blood. 2009;114:4441–50.

    Article  CAS  Google Scholar 

  8. Ten Hacken E, Burger JA. Microenvironment interactions and B-cell receptor signaling in chronic lymphocytic leukemia: implications for disease pathogenesis and treatment. Biochim Biophys Acta. 2016;1863:401–13.

    Article  Google Scholar 

  9. Jin L, Liu WR, Tian MX, Fan J, Shi YH. The SphKs/S1P/S1PR1 axis in immunity and cancer: more ore to be mined. World J Surg Oncol. 2016;14:131.

    Article  Google Scholar 

  10. Redondo-Munoz J, Jose Terol M, Garcia-Marco JA, Garcia-Pardo A. Matrix metalloproteinase-9 is up-regulated by CCL21/CCR7 interaction via extracellular signal-regulated kinase-1/2 signaling and is involved in CCL21-driven B-cell chronic lymphocytic leukemia cell invasion and migration. Blood. 2008;111:383–6.

    Article  CAS  Google Scholar 

  11. Patrussi L, Capitani N, Martini V, Pizzi M, Trimarco V, Frezzato F, et al. Enhanced chemokine receptor recycling and impaired S1P1 expression promote leukemic cell infiltration of lymph nodes in chronic lymphocytic leukemia. Cancer Res. 2015;75:4153–63.

    Article  CAS  Google Scholar 

  12. López-Giral S, Quintana NE, Cabrerizo M, Alfonso-Pérez M, Sala-Valdés M, De Soria VG, et al. Chemokine receptors that mediate B cell homing to secondary lymphoid tissues are highly expressed in B cell chronic lymphocytic leukemia and non-Hodgkin lymphomas with widespread nodular dissemination. J Leukoc Biol. 2004;76:462–71.

    Article  Google Scholar 

  13. Capitani N, Patrussi L, Trentin L, Lucherini OM, Cannizzaro E, Migliaccio E, et al. S1P1 expression is controlled by the pro-oxidant activity of p66Shc and is impaired in B-CLL patients with unfavorable prognosis. Blood. 2012;120:4391–9.

    Article  CAS  Google Scholar 

  14. Kang DS, Tian X, Benovic JL. Role of beta-arrestins and arrestin domain-containing proteins in G protein-coupled receptor trafficking. Curr Opin Cell Biol. 2014;27:63–71.

    Article  Google Scholar 

  15. Gurevich VV, Gurevich EV. Arrestins: critical players in trafficking of many GPCRs. Prog Mol Biol Transl Sci. 2015;132:1–14.

    Article  CAS  Google Scholar 

  16. Marchese A. Endocytic trafficking of chemokine receptors. Curr Opin Cell Biol. 2014;27:72–77.

    Article  CAS  Google Scholar 

  17. Otero C, Groettrup M, Legler DF. Opposite fate of endocytosed CCR7 and its ligands: recycling versus degradation. J Immunol. 2006;177:2314–23.

    Article  CAS  Google Scholar 

  18. Zhang Y, Foudi A, Geay JF, Berthebaud M, Buet D, Jarrier P, et al. Intracellular localization and constitutive endocytosis of CXCR4 in human CD34+hematopoietic progenitor cells. Stem Cells. 2004;22:1015–29.

    Article  CAS  Google Scholar 

  19. Capitani N, Lucherini OM, Sozzi E, Ferro M, Giommoni N, Finetti F, et al. Impaired expression of p66Shc, a novel regulator of B-cell survival, in chronic lymphocytic leukemia. Blood. 2010;115:3726–36.

    Article  CAS  Google Scholar 

  20. Patrussi L, Capitani N, Cannizzaro E, Finetti F, Lucherini OM, Pelicci PG, et al. Negative regulation of chemokine receptor signaling and B-cell chemotaxis by p66Shc. Cell Death Dis. 2014;5:e1068.

    Article  CAS  Google Scholar 

  21. Vasudevan NT, Mohan ML, Goswami SK, Naga Prasad SV. Regulation of beta-adrenergic receptor function: an emphasis on receptor resensitization. Cell Cycle. 2011;10:3684–91.

    Article  CAS  Google Scholar 

  22. Bolinger MT, Ramshekar A, Waldschmidt HV, Larsen SD, Bewley MC, Flanagan JM, et al. Occludin S471 phosphorylation contributes to epithelial monolayer maturation. Mol Cell Biol. 2016;36:2051–66.

    Article  CAS  Google Scholar 

  23. Zhou T, Song L, Yang P, Wang Z, Lui D, Jope RS. Bisindolylmaleimide VIII facilitates Fas-mediated apoptosis and inhibits T cell-mediated autoimmune diseases. Nat Med. 1999;5:42–48.

    Article  CAS  Google Scholar 

  24. Murray AJ. Pharmacological PKA inhibition: all may not be what it seems. Sci Signal. 2008;1:re4.

    Article  Google Scholar 

  25. Swingle M, Ni L, Honkanen RE. Small-molecule inhibitors of ser/thr protein phosphatases: specificity, use and common forms of abuse. Methods Mol Biol. 2007;365:23–38.

    PubMed  PubMed Central  Google Scholar 

  26. Martinez-Martinez S, Redondo JM. Inhibitors of the calcineurin/NFAT pathway. Curr Med Chem. 2004;11:997–1007.

    Article  CAS  Google Scholar 

  27. Schulz RA, Yutzey KE. Calcineurin signaling and NFAT activation in cardiovascular and skeletal muscle development. Dev Biol. 2004;266:1–16.

    Article  CAS  Google Scholar 

  28. Serfling E, Berberich-Siebelt F, Chuvpilo S, Jankevics E, Klein-Hessling S, Twardzik T, et al. The role of NF-AT transcription factors in T cell activation and differentiation. Biochim Biophys Acta. 2000;1498:1–18.

    Article  CAS  Google Scholar 

  29. Burger JA, Tedeschi A, Barr PM, Robak T, Owen C, Ghia P, et al. Ibrutinib as initial therapy for patients with chronic lymphocytic leukemia. N Engl J Med. 2015;373:2425–37.

    Article  CAS  Google Scholar 

  30. Lee CS, Rattu MA, Kim SS. A review of a novel, Bruton’s tyrosine kinase inhibitor, ibrutinib. J Oncol Pharm Pract. 2016;22:92–104.

    Article  Google Scholar 

  31. Byrd JC, Furman RR, Coutre SE, Flinn IW, Burger JA, Blum KA, et al. Targeting BTK with ibrutinib in relapsed chronic lymphocytic leukemia. N Engl J Med. 2013;369:32–42.

    Article  CAS  Google Scholar 

  32. Cheng S, Ma J, Guo A, Lu P, Leonard JP, Coleman M, et al. BTK inhibition targets in vivo CLL proliferation through its effects on B-cell receptor signaling activity. Leukemia. 2014;28:649–57.

    Article  CAS  Google Scholar 

  33. Wodarz D, Garg N, Komarova NL, Benjamini O, Keating MJ, Wierda WG, et al. Kinetics of CLL cells in tissues and blood during therapy with the BTK inhibitor ibrutinib. Blood. 2014;123:4132–5.

    Article  CAS  Google Scholar 

  34. de Rooij MF, Kuil A, Geest CR, Eldering E, Chang BY, Buggy JJ, et al. The clinically active BTK inhibitor PCI-32765 targets B-cell receptor- and chemokine-controlled adhesion and migration in chronic lymphocytic leukemia. Blood. 2012;119:2590–4.

    Article  Google Scholar 

  35. Hjortø GM, Larsen O, Steen A, Daugvilaite V, Berg C, Fares S, et al. Differential CCR7 targeting in dendritic cells by three naturally occurring CC-chemokines. Front Immunol. 2016;7:568.

    Article  Google Scholar 

  36. Otero C, Eisele PS, Schaeuble K, Groettrup M, Legler DF. Distinct motifs in the chemokine receptor CCR7 regulate signal transduction, receptor trafficking and chemotaxis. J Cell Sci. 2008;121:2759–67.

    Article  CAS  Google Scholar 

  37. Hauser MA, Legler DF. Common and biased signaling pathways of the chemokine receptor CCR7 elicited by its ligands CCL19 and CCL21 in leukocytes. J Leukoc Biol. 2016;99:869–82.

    Article  CAS  Google Scholar 

  38. Charest-Morin X, Pepin R, Gagne-Henley A, Morissette G, Lodge R, Marceau F, C-C chemokine receptor-7 mediated endocytosis of antibody cargoes into intact cells. Front Pharmacol. 2013;4:122.

    Article  Google Scholar 

  39. Lagane B, Chow KY, Balabanian K, Levoye A, Harriague J, Planchenault T, et al. CXCR4 dimerization and beta-arrestin-mediated signaling account for the enhanced chemotaxis to CXCL12 in WHIM syndrome. Blood. 2008;112:34–44.

    Article  CAS  Google Scholar 

  40. Bamidele AO, Kremer KN, Hirsova P, Clift IC, Gores GJ, Billadeau DD, et al. IQGAP1 promotes CXCR4 chemokine receptor function and trafficking via EEA-1+endosomes. J Cell Biol. 2015;210:257–72.

    Article  CAS  Google Scholar 

  41. Kennedy JE, Marchese A. Regulation of GPCR trafficking by ubiquitin. Prog Mol Biol Transl Sci. 2015;132:15–38.

    Article  CAS  Google Scholar 

  42. Berlin I, Higginbotham KM, Dise RS, Sierra MI, Nash PD. The deubiquitinating enzyme USP8 promotes trafficking and degradation of the chemokine receptor 4 at the sorting endosome. J Biol Chem. 2010;285:37895–908.

    Article  CAS  Google Scholar 

  43. Seachrist JL, Ferguson SS. Regulation of G protein-coupled receptor endocytosis and trafficking by Rab GTPases. Life Sci. 2003;74:225–35.

    Article  CAS  Google Scholar 

  44. Seachrist JL, Laporte SA, Dale LB, Babwah AV, Caron MG, Anborgh PH, et al. Rab5 association with the angiotensin II type 1A receptor promotes Rab5 GTP binding and vesicular fusion. J Biol Chem. 2002;277:679–85.

    Article  CAS  Google Scholar 

  45. van IJzendoorn SC, Tuvim MJ, Weimbs T, Dickey BF, Mostov KE. Direct interaction between Rab3b and the polymeric immunoglobulin receptor controls ligand-stimulated transcytosis in epithelial cells. Dev Cell. 2002;2:219–28.

    Article  Google Scholar 

  46. Li L, Baxter SS, Gu N, Ji M, Zhan X. Missing-in-metastasis protein downregulates CXCR4 by promoting ubiquitylation and interaction with small Rab GTPases. J Cell Sci. 2017;130:1475–85.

    Article  CAS  Google Scholar 

  47. Wienands J. The B-cell antigen receptor: formation of signaling complexes and the function of adaptor proteins. Curr Top Microbiol Immunol. 2000;245:53–76.

    CAS  PubMed  Google Scholar 

  48. Bürkle A, Niedermeier M, Schmitt-Gräff A, Wierda WG, Keating MJ, Burger JA. Overexpression of the CXCR5 chemokine receptor, and its ligand, CXCL13 in B-cell chronic lymphocytic leukemia. Blood. 2007;110:3316–25.

    Article  Google Scholar 

  49. Calebiro D, Nikolaev VO, Persani L, Lohse MJ. Signaling by internalized G-protein-coupled receptors. Trends Pharmacol Sci. 2010;31:221–8.

    Article  CAS  Google Scholar 

  50. Bowman SL, Shiwarski DJ, Puthenveedu MA. Distinct G protein-coupled receptor recycling pathways allow spatial control of downstream G protein signaling. J Cell Biol. 2016;214:797–806.

    Article  CAS  Google Scholar 

  51. Herman SE, Mustafa RZ, Jones J, Wong DH, Farooqui M, Wiestner A. Treatment with ibrutinib inhibits BTK- and VLA-4-dependent adhesion of chronic lymphocytic leukemia cells in vivo. Clin Cancer Res. 2015;21:4642–51.

    Article  CAS  Google Scholar 

  52. Cattaneo F, Patrussi L, Capitani N, Frezzato F, D’Elios MM, Trentin L, et al. Expression of the p66Shc protein adaptor is regulated by the activator of transcription STAT4 in normal and chronic lymphocytic leukemia B cells. Oncotarget. 2016;7:57086–98.

    Article  Google Scholar 

  53. Visentin A, Facco M, Frezzato F, Castelli M, Trimarco V, Martini V, et al. Integrated CLL scoring system, a new and simple index to predict time to treatment and overall survival in patients with chronic lymphocytic leukemia. Clin Lymphoma Myeloma Leuk. 2015;15:612–20.

    Article  Google Scholar 

  54. Stacchini A, Aragno M, Vallario A, Alfarano A, Circosta P, Gottardi D, et al. MEC1 and MEC2: two new cell lines derived from B-chronic lymphocytic leukaemia in prolymphocytoid transformation. Leuk Res. 1999;23:127–36.

    Article  CAS  Google Scholar 

  55. Migliaccio E, Giorgio M, Mele S, Pelicci G, Reboldi P, Pandolfi PP, et al. The p66shc adaptor protein controls oxidative stress response and life span in mammals. Nature. 1999;402:309–13.

    Article  CAS  Google Scholar 

  56. Patrussi L, Ulivieri C, Lucherini OM, Paccani SR, Gamberucci A, Lanfrancone L, et al. p52Shc is required for CXCR4-dependent signaling and chemotaxis in T cells. Blood. 2007;110:1730–8.

    Article  CAS  Google Scholar 

  57. Patrussi L, Baldari CT. Analysis of TCR/CD3 recycling at the immune synapse. Methods Mol Biol. 2017;1584:143–55.

    Article  CAS  Google Scholar 

  58. Gamberucci A, Giunti R, Benedetti A. Progesterone inhibits capacitative Ca2+ entry in Jurkat T lymphocytes by a membrane delimited mechanism, independently of plasma membrane depolarization. Cell Calcium. 2004;36:175–80.

    Article  CAS  Google Scholar 

  59. Finetti F, Paccani SR, Riparbelli MG, Giacomello E, Perinetti G, Pazour GJ, et al. Intraflagellar transport is required for polarized recycling of the TCR/CD3 complex to the immune synapse. Nat Cell Biol. 2009;11:1332–9.

    Article  CAS  Google Scholar 

  60. Manders EM, Stap J, Brakenhoff GJ, van Driel R, Aten JA. Dynamics of three-dimensional replication patterns during the S-phase, analysed by double labelling of DNA and confocal microscopy. J Cell Sci. 1992;103:857–62.

    CAS  PubMed  Google Scholar 

  61. Plyte S, Boncristiano M, Fattori E, Galvagni F, Paccani SR, Majolini MB, et al. Identification and characterization of a novel nuclear factor of activated T-cells-1 isoform expressed in mouse brain. J Biol Chem. 2001;276:14350–8.

    Article  CAS  Google Scholar 

  62. Boncristiano M, Paccani SR, Barone S, Ulivieri C, Patrussi L, Ilver D, et al. The Helicobacter pylori vacuolating toxin inhibits T cell activation by two independent mechanisms. J Exp Med. 2003;198:1887–97.

    Article  CAS  Google Scholar 

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Acknowledgements

The authors wish to thank Sonia Grassini for technical assistance.

Funding

This work was carried out with the support of grant AIRC IG-15220 and ITT-Regione Toscana to CTB, AIRC IG-15286, Cariparo and Cariverona to GS and AIRC IG-15397 to LT.

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Author notes

  1. Laura Patrussi and Nagaja Capitani contributed equally to this work.

Authors and Affiliations

  1. Department of Life Sciences, University of Siena, Siena, Italy

    Laura Patrussi, Nagaja Capitani, Francesca Cattaneo, Noemi Manganaro & Cosima T. Baldari

  2. Department of Clinical and Experimental Medicine, University of Florence, Florence, Italy

    Nagaja Capitani & Mario M. D’Elios

  3. Department of Molecular and Developmental Medicine, University of Siena, Siena, Italy

    Alessandra Gamberucci

  4. Venetian Institute of Molecular Medicine, Padua, Italy

    Federica Frezzato, Veronica Martini, Andrea Visentin, Livio Trentin & Gianpietro Semenzato

  5. Department of Medicine, Hematology and Clinical Immunology Branch, Padua University School of Medicine, Padua, Italy

    Federica Frezzato, Veronica Martini, Andrea Visentin, Livio Trentin & Gianpietro Semenzato

  6. European Institute of Oncology, Milan, Italy

    Pier Giuseppe Pelicci

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Patrussi, L., Capitani, N., Cattaneo, F. et al. p66Shc deficiency enhances CXCR4 and CCR7 recycling in CLL B cells by facilitating their dephosphorylation-dependent release from β-arrestin at early endosomes. Oncogene 37, 1534–1550 (2018). https://doi.org/10.1038/s41388-017-0066-2

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