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GBM-Derived Wnt3a Induces M2-Like Phenotype in Microglial Cells Through Wnt/β-Catenin Signaling

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Abstract

Glioblastoma is an extremely aggressive and deadly brain tumor known for its striking cellular heterogeneity and capability to communicate with microenvironment components, such as microglia. Microglia-glioblastoma interaction contributes to an increase in tumor invasiveness, and Wnt signaling pathway is one of the main cascades related to tumor progression through changes in cell migration and invasion. However, very little is known about the role of canonical Wnt signaling during microglia-glioblastoma crosstalk. Here, we show for the first time that Wnt3a is one of the factors that regulate interactions between microglia and glioblastoma cells. Wnt3a activates the Wnt/β-catenin signaling of both glioblastoma and microglial cells. Glioblastoma-conditioned medium not only induces nuclear translocation of microglial β-catenin but also increases microglia viability and proliferation as well as Wnt3a, cyclin-D1, and c-myc expression. Moreover, glioblastoma-derived Wnt3a increases microglial ARG-1 and STI1 expression, followed by an upregulation of IL-10 mRNA levels, and a decrease in IL1β gene expression. The presence of Wnt3a in microglia-glioblastoma co-cultures increases the formation of membrane nanotubes accompanied by changes in migration capability. In vivo, tumors formed from Wnt3a-stimulated glioblastoma cells presented greater microglial infiltration and more aggressive characteristics such as growth rate than untreated tumors. Thus, we propose that Wnt3a belongs to the arsenal of factors capable of stimulating the induction of M2-like phenotype on microglial cells, which contributes to the poor prognostic of glioblastoma, reinforcing that Wnt/β-catenin pathway can be a potential therapeutic target to attenuate glioblastoma progression.

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References

  1. Stupp R, Hegi ME, Gilbert MR, Chakravarti A (2007) Chemoradiotherapy in malignant glioma: standard of care and future directions. J Clin Oncol 25:4127–4136. https://doi.org/10.1200/JCO.2007.11.8554

    Article  CAS  PubMed  Google Scholar 

  2. Louis DN, Ohgaki H, Wiestler OD, Cavenee WK, Burger PC, Jouvet A, Scheithauer BW, Kleihues P (2007) The 2007 WHO classification of tumours of the central nervous system. Acta Neuropathol 114:97–109. https://doi.org/10.1007/s00401-007-0243-4

    Article  PubMed  PubMed Central  Google Scholar 

  3. Friedmann-Morvinski D (2014) Glioblastoma heterogeneity and cancer cell plasticity. Crit Rev Oncog 19:327–336

    Article  PubMed  Google Scholar 

  4. Albini A, Bruno A, Gallo C, Pajardi G, Noonan DM, Dallaglio K (2015) Cancer stem cells and the tumor microenvironment: interplay in tumor heterogeneity. Connect Tissue Res 56:414–425. https://doi.org/10.3109/03008207.2015.1066780

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Balça-Silva J, Matias D, Do Carmo A, Dubois LG, Gonçalves AC, Girão H, Silva Canedo NH, Correia AH et al (2017) Glioblastoma entities express subtle differences in molecular composition and response to treatment. Oncol Rep 38:1341–1352. https://doi.org/10.3892/or.2017.5799

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Abou-Antoun TJ, Hale JS, Lathia JD, Dombrowski SM (2017) Brain cancer stem cells in adults and children: cell biology and therapeutic implications. Neurotherapeutics 14:372–384. https://doi.org/10.1007/s13311-017-0524-0

    Article  PubMed  PubMed Central  Google Scholar 

  7. Roos A, Ding Z, Loftus JC, Tran NL (2017) Molecular and microenvironmental determinants of glioma stem-like cell survival and invasion. Front Oncol 7:120. https://doi.org/10.3389/fonc.2017.00120

    Article  PubMed  PubMed Central  Google Scholar 

  8. Garcia C, Dubois LG, Xavier AL, Geraldo LH, da Fonseca ACC, Correia AH, Meirelles F, Ventura G et al (2014) The orthotopic xenotransplant of human glioblastoma successfully recapitulates glioblastoma-microenvironment interactions in a non-immunosuppressed mouse model. BMC Cancer 14:923. https://doi.org/10.1186/1471-2407-14-923

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Carvalho da Fonseca AC, Wang H, Fan H, Chen X, Zhang I, Zhang L, Lima FRS, Badie B (2014) Increased expression of stress inducible protein 1 in glioma-associated microglia/macrophages. J Neuroimmunol 274:71–77. https://doi.org/10.1016/j.jneuroim.2014.06.021

    Article  CAS  PubMed  Google Scholar 

  10. Olah M, Raj D, Brouwer N, de Haas AH, Eggen BJL, den Dunnen WFA, Biber KPH, Boddeke HWGM (2012) An optimized protocol for the acute isolation of human microglia from autopsy brain samples. Glia 60:96–111. https://doi.org/10.1002/glia.21251

    Article  PubMed  Google Scholar 

  11. Razavi S-M, Lee KE, Jin BE, Aujla PS, Gholamin S, Li G (2016) Immune evasion strategies of glioblastoma. Front Surg 3:11. https://doi.org/10.3389/fsurg.2016.00011

    Article  PubMed  PubMed Central  Google Scholar 

  12. da Fonseca ACC, Amaral R, Garcia C et al (2016) Microglia in cancer: for good or for bad? Adv Exp Med Biol 949:245–261. https://doi.org/10.1007/978-3-319-40764-7_12

    Article  CAS  PubMed  Google Scholar 

  13. Audia A, Conroy S, Glass R, Bhat KPL (2017) The impact of the tumor microenvironment on the properties of glioma stem-like cells. Front Oncol 7:143. https://doi.org/10.3389/fonc.2017.00143

    Article  PubMed  PubMed Central  Google Scholar 

  14. Van Meir E, Sawamura Y, Diserens AC et al (1990) Human glioblastoma cells release interleukin 6 in vivo and in vitro. Cancer Res 50:6683–6688

    PubMed  Google Scholar 

  15. Woiciechowsky C, Asadullah K, Nestler D, Glockner F, Robinson PN, Volk HD, Vogel S, Lanksch WR (1997) Different release of cytokines into the cerebrospinal fluid following surgery for intra- and extra-axial brain tumours. Acta Neurochir 139:619–624

    Article  CAS  PubMed  Google Scholar 

  16. Constam DB, Philipp J, Malipiero UV et al (1992) Differential expression of transforming growth factor-beta 1, -beta 2, and -beta 3 by glioblastoma cells, astrocytes, and microglia. J Immunol 148:1404–1410

    CAS  PubMed  Google Scholar 

  17. Hambardzumyan D, Gutmann DH, Kettenmann H (2016) The role of microglia and macrophages in glioma maintenance and progression. Nat Neurosci 19:20–27. https://doi.org/10.1038/nn.4185

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Markovic DS, Glass R, Synowitz M, Rooijen N, Kettenmann H (2005) Microglia stimulate the invasiveness of glioma cells by increasing the activity of metalloprotease-2. J Neuropathol Exp Neurol 64:754–762

    Article  CAS  PubMed  Google Scholar 

  19. Matias D, Predes D, Niemeyer Filho P, Lopes MC, Abreu JG, Lima FRS, Moura Neto V (2017) Microglia-glioblastoma interactions: new role for Wnt signaling. Biochim Biophys Acta 1868:333–340. https://doi.org/10.1016/j.bbcan.2017.05.007

    Article  CAS  Google Scholar 

  20. Zhang I, Alizadeh D, Liang J, Zhang L, Gao H, Song Y, Ren H, Ouyang M et al (2016) Characterization of arginase expression in glioma-associated microglia and macrophages. PLoS One 11:e0165118. https://doi.org/10.1371/journal.pone.0165118

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Fonseca ACC, da Romão L, Amaral RF et al (2012) Microglial stress inducible protein 1 promotes proliferation and migration in human glioblastoma cells. Neuroscience 200:130–141. https://doi.org/10.1016/j.neuroscience.2011.10.025

    Article  CAS  PubMed  Google Scholar 

  22. Denysenko T, Annovazzi L, Cassoni P et al (2016) WNT/β-catenin signaling pathway and downstream modulators in low- and high-grade glioma. Cancer Genomics Proteomics 13:31–45

    CAS  PubMed  Google Scholar 

  23. Halleskog C, Mulder J, Dahlström J, Mackie K, Hortobágyi T, Tanila H, Kumar Puli L, Färber K et al (2011) WNT signaling in activated microglia is proinflammatory. Glia 59:119–131. https://doi.org/10.1002/glia.21081

    Article  PubMed  Google Scholar 

  24. Zheng H, Jia L, Liu C-C, Rong Z, Zhong L, Yang L, Chen XF, Fryer JD et al (2017) TREM2 promotes microglial survival by activating Wnt/β-catenin pathway. J Neurosci 37:1772–1784. https://doi.org/10.1523/JNEUROSCI.2459-16.2017

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Dijksterhuis JP, Arthofer E, Marinescu VD, Nelander S, Uhlén M, Pontén F, Mulder J, Schulte G (2015) High levels of WNT-5A in human glioma correlate with increased presence of tumor-associated microglia/monocytes. Exp Cell Res 339:280–288. https://doi.org/10.1016/j.yexcr.2015.10.022

    Article  CAS  PubMed  Google Scholar 

  26. Faria J, Romão L, Martins S, Alves T, Mendes FA, de Faria GP, Hollanda R, Takiya C et al (2006) Interactive properties of human glioblastoma cells with brain neurons in culture and neuronal modulation of glial laminin organization. Differentiation 74:562–572. https://doi.org/10.1111/j.1432-0436.2006.00090.x

    Article  CAS  PubMed  Google Scholar 

  27. Kahn SA, Biasoli D, Garcia C et al (2012) Equinatoxin II potentiates temozolomide- and etoposide-induced glioblastoma cell death. Curr Top Med Chem 12:2082–2093

    Article  CAS  PubMed  Google Scholar 

  28. Lima FR, Gervais A, Colin C et al (2001) Regulation of microglial development: a novel role for thyroid hormone. J Neurosci 21:2028–2038

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Towbin H, Staehelin T, Gordon J (1992) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. 1979 Biotechnology 24:145–149

    CAS  PubMed  Google Scholar 

  30. Amado NG, Fonseca BF, Cerqueira DM, Neto VM, Abreu JG (2011) Flavonoids: potential Wnt/beta-catenin signaling modulators in cancer. Life Sci 89:545–554. https://doi.org/10.1016/j.lfs.2011.05.003

    Article  CAS  PubMed  Google Scholar 

  31. Oloumi A, Syam S, Dedhar S (2006) Modulation of Wnt3a-mediated nuclear beta-catenin accumulation and activation by integrin-linked kinase in mammalian cells. Oncogene 25:7747–7757. https://doi.org/10.1038/sj.onc.1209752

    Article  CAS  PubMed  Google Scholar 

  32. Liao DJ, Thakur A, Wu J, Biliran H, Sarkar FH (2007) Perspectives on c-Myc, cyclin D1, and their interaction in cancer formation, progression, and response to chemotherapy. Crit Rev Oncog 13:93–158

    Article  PubMed  Google Scholar 

  33. Clément-Lacroix P, Ai M, Morvan F et al (2005) Lrp5-independent activation of Wnt signaling by lithium chloride increases bone formation and bone mass in mice. Proc Natl Acad Sci U S A 102:17406–17411. https://doi.org/10.1073/pnas.0505259102

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Ajmone-Cat MA, D’Urso MC, di Blasio G, Brignone MS, de Simone R, Minghetti L (2016) Glycogen synthase kinase 3 is part of the molecular machinery regulating the adaptive response to LPS stimulation in microglial cells. Brain Behav Immun 55:225–235. https://doi.org/10.1016/j.bbi.2015.11.012

    Article  CAS  PubMed  Google Scholar 

  35. Crain JM, Nikodemova M, Watters JJ (2013) Microglia express distinct M1 and M2 phenotypic markers in the postnatal and adult central nervous system in male and female mice. J Neurosci Res 91:1143–1151. https://doi.org/10.1002/jnr.23242

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Dey N, Barwick BG, Moreno CS, Ordanic-Kodani M, Chen Z, Oprea-Ilies G, Tang W, Catzavelos C et al (2013) Wnt signaling in triple negative breast cancer is associated with metastasis. BMC Cancer 13:537. https://doi.org/10.1186/1471-2407-13-537

    Article  PubMed  PubMed Central  Google Scholar 

  37. Yamaguchi H, Condeelis J (2007) Regulation of the actin cytoskeleton in cancer cell migration and invasion. Biochim Biophys Acta 1773:642–652. https://doi.org/10.1016/j.bbamcr.2006.07.001

    Article  CAS  PubMed  Google Scholar 

  38. Shibamoto S, Higano K, Takada R, Ito F, Takeichi M, Takada S (1998) Cytoskeletal reorganization by soluble Wnt-3a protein signalling. Genes Cells 3:659–670

    Article  CAS  PubMed  Google Scholar 

  39. Holmes KC (2009) Structural biology: actin in a twist. Nature 457:389–390. https://doi.org/10.1038/457389a

    Article  CAS  PubMed  Google Scholar 

  40. Kaur N, Chettiar S, Rathod S, Rath P, Muzumdar D, Shaikh ML, Shiras A (2013) Wnt3a mediated activation of Wnt/β-catenin signaling promotes tumor progression in glioblastoma. Mol Cell Neurosci 54:44–57. https://doi.org/10.1016/j.mcn.2013.01.001

    Article  CAS  PubMed  Google Scholar 

  41. Zhang Z-M, Yang Z, Zhang Z (2015) Distribution and characterization of tumor-associated macrophages/microglia in rat C6 glioma. Oncol Lett 10:2442–2446. https://doi.org/10.3892/ol.2015.3533

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. O’Connor T, Borsig L, Heikenwalder M (2015) CCL2-CCR2 signaling in disease pathogenesis. Endocr Metab Immune Disord Drug Targets 15:105–118

    Article  PubMed  Google Scholar 

  43. Pontes B, Viana NB, Campanati L, Farina M, Neto VM, Nussenzveig HM (2008) Structure and elastic properties of tunneling nanotubes. Eur Biophys J 37:121–129. https://doi.org/10.1007/s00249-007-0184-9

    Article  CAS  PubMed  Google Scholar 

  44. Hooper C, Sainz-Fuertes R, Lynham S, Hye A, Killick R, Warley A, Bolondi C, Pocock J et al (2012) Wnt3a induces exosome secretion from primary cultured rat microglia. BMC Neurosci 13:144. https://doi.org/10.1186/1471-2202-13-144

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Nawaz M, Fatima F (2017) Extracellular vesicles, tunneling nanotubes, and cellular interplay: synergies and missing links. Front Mol Biosci 4:50. https://doi.org/10.3389/fmolb.2017.00050

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This study was supported by the Brazilian agencies Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ), Pró-Saúde Associação Beneficente de Assistência Social e Hospitalar, and Ary Frauzino Foundation for Cancer Research.

We would like to acknowledge Dra. Juliana Coelho Aguiar for giving us some primers, Dra. Graziella Ventura for helping us with the confocal microscopy acquisitions, and Geralda Cardoso for the lab technical support.

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Authors and Affiliations

  1. Laboratório de Biomedicina do Cérebro, Instituto Estadual do Cérebro Paulo Niemeyer, Secretaria de Saúde do Estado do Rio de Janeiro, Rua do Resende 156, Rio de Janeiro, CEP 20231-092, Brazil

    Diana Matias, Luiz Gustavo Dubois, Luciane Rosário, Valeria Pereira Ferrer, Joana Balça-Silva, Lucy Wanjiku Macharia, Tania Cristina Leite de Sampaio e Spohr, Leila Chimelli, Paulo Niemeyer Filho & Vivaldo Moura-Neto

  2. Instituto de Ciências Biomédicas da Universidade Federal do Rio de Janeiro (ICB/UFRJ), Rio de Janeiro, 21941-902, Brazil

    Diana Matias, Luiz Gustavo Dubois, Bruno Pontes, Anna Carolina Carvalho Fonseca, Luciana Romão, José Garcia Abreu & Flavia Regina Souza Lima

  3. Centro de Neurociências e Biologia celular e Instituto Biomédico da Imagem e das Ciências da Vida (CNC.IBILI), Coimbra, Portugal

    Joana Balça-Silva & Maria Celeste Lopes

  4. Faculdade de Medicina da Universidade de Coimbra (FMUC), Coimbra, Portugal

    Joana Balça-Silva

  5. Pólo das Ciências da Saúde, Faculdade de Farmácia da Universidade de Coimbra, Coimbra, Portugal

    Maria Celeste Lopes

  6. Campus Duque de Caxias, Universidade Federal do Rio de Janeiro, Duque de Caxias, Brazil

    Luciana Romão

  7. Programa de Pós-Graduação em Anatomia Patológica, Faculdade de Medicina da Universidade Federal do Rio de Janeiro -UFRJ, Rio de Janeiro, Brazil

    Luciane Rosário & Lucy Wanjiku Macharia

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  1. Diana Matias
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Correspondence to Vivaldo Moura-Neto.

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Matias, D., Dubois, L.G., Pontes, B. et al. GBM-Derived Wnt3a Induces M2-Like Phenotype in Microglial Cells Through Wnt/β-Catenin Signaling. Mol Neurobiol 56, 1517–1530 (2019). https://doi.org/10.1007/s12035-018-1150-5

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