Abstract
Objective and design
We sought to determine the effect of necrosis-induced activation of the complement protein C3 in medulloblastoma.
Materials/methods
Twelve medulloblastoma surgical specimens were evaluated for complement activation using immunohistochemistry, with H&E stains performed on adjacent tissue sections to determine the relationship of complement activation to necrotic tissue. Flow cytometry and Western blot were performed on three established medulloblastoma lines and one surgically-procured cell culture to determine expression of C3a receptor (C3aR) in medulloblastoma. In vitro proliferation of siRNA C3aR knockdown cells was compared to that of control siRNA cells with cell line Daoy.
Results
Three surgical specimens were found to have necrosis on H&E sections. In each case, iC3b staining was identified on adjacent sections, limited to the necrotic region. In no case did necrosis occur without iC3b staining on adjacent sections. C3aR protein was demonstrated on both the three established cell lines and on the surgical culture. Proliferation assays of Daoy cells with siRNA knockdown vs. control siRNA revealed significantly reduced proliferation at 72 h (p = 0.001).
Conclusions
Necrosis is associated with complement activation in medulloblastoma. Medulloblastoma cells express C3aR, and siRNA-mediated knockdown of C3aR inhibits proliferation of these cells in vitro.
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References
Proctor LM, Arumugam TV, Shiels I, Reid RC, Fairlie DP, Taylor SM. Comparative anti-inflammatory activities of antagonists to C3a and C5a receptors in a rat model of intestinal ischaemia/reperfusion injury. Br J Pharmacol. 2004;142:756–64.
Elsner J, Oppermann M, Czech W, Kapp A. C3a activates the respiratory burst in human polymorphonuclear neutrophilic leukocytes via pertussis toxin-sensitive G-proteins. Blood. 1994;83:3324–31.
Walport MJ. Complement. First of two parts. N Engl J Med. 2001;344:1058–66.
Walport MJ. Complement. Second of two parts. N Engl J Med. 2001;344:1140–4.
Gasque P, Singhrao SK, Neal JW, Gotze O, Morgan BP. Expression of the receptor for complement C5a (CD88) is up-regulated on reactive astrocytes, microglia, and endothelial cells in the inflamed human central nervous system. Am J Pathol. 1997;150:31–41.
Gasque P, Dean YD, McGreal EP, VanBeek J, Morgan BP. Complement components of the innate immune system in health and disease in the CNS. Immunopharmacology. 2000;49:171–86.
Ciurana CL, Zwart B, van Mierlo G, Hack CE. Complement activation by necrotic cells in normal plasma environment compares to that by late apoptotic cells and involves predominantly IgM. Eur J Immunol. 2004;34:2609–19.
Janeway CA, Travers P, Walport M, Capra JD. Immunobiology: the immune system in health and disease. London: Churchill Livingstone; 2001.
Markiewski MM, Lambris JD. The role of complement in inflammatory diseases from behind the scenes into the spotlight. Am J Pathol. 2007;171:715–27.
Mastellos D, Lambris JD. Complement: more than a ‘guard’ against invading pathogens? Trends Immunol. 2002;23:485–91.
Rutkowski MJ, Sughrue ME, Kane AJ, Ahn BJ, Fang S, Parsa AT. The complement cascade as a mediator of tissue growth and regeneration. Inflamm Res. 2010;59:897–905.
Rutkowski MJ, Sughrue ME, Kane AJ, Mills SA, Fang S, Parsa AT. Complement and the central nervous system: emerging roles in development, protection and regeneration. Immunol Cell Biol. 2010;88:781–6.
Gaipl US, Kuenkele S, Voll RE, Beyer TD, Kolowos W, Heyder P, et al. Complement binding is an early feature of necrotic and a rather late event during apoptotic cell death. Cell Death Differ. 2001;8:327–34.
Gasque P, Neal JW, Singhrao SK, McGreal EP, Dean YD, Van BJ, et al. Roles of the complement system in human neurodegenerative disorders: pro-inflammatory and tissue remodeling activities. Mol Neurobiol. 2002;25:1–17.
Humbles AA, Lu B, Nilsson CA, Lilly C, Israel E, Fujiwara Y, et al. A role for the C3a anaphylatoxin receptor in the effector phase of asthma. Nature. 2000;406:998–1001.
Mathieu MC, Sawyer N, Greig GM, Hamel M, Kargman S, Ducharme Y, et al. The C3a receptor antagonist SB 290157 has agonist activity. Immunol Lett. 2005;100:139–45.
Qu H, Magotti P, Ricklin D, Wu EL, Kourtzelis I, Wu YQ, et al. Novel analogues of the therapeutic complement inhibitor compstatin with significantly improved affinity and potency. Mol Immunol. 2011;48:481–9.
Rahpeymai Y, Hietala MA, Wilhelmsson U, Fotheringham A, Davies I, Nilsson AK, et al. Complement: a novel factor in basal and ischemia-induced neurogenesis. EMBO J. 2006;25:1364–74.
Ricklin D, Lambris JD. Compstatin: a complement inhibitor on its way to clinical application. Adv Exp Med Biol. 2008;632:273–92.
Speth C, Dierich MP, Gasque P. Neuroinvasion by pathogens: a key role of the complement system. Mol Immunol. 2002;38:669–79.
Rutkowski MJ, Sughrue ME, Kane AJ, Mills SA, Parsa AT. Cancer and the complement cascade. Mol Cancer Res. 2010;8:1453–65.
Gasque P, Thomas A, Fontaine M, Morgan BP. Complement activation on human neuroblastoma cell lines in vitro: route of activation and expression of functional complement regulatory proteins. J Neuroimmunol. 1996;66:29–40.
Kaiser HE, Bodey B. The role of apoptosis in normal ontogenesis and solid human neoplasms. In vivo Athens Greece. 2000;14:789–803.
Fishelson Z, Donin N, Zell S, Schultz S, Kirschfink M. Obstacles to cancer immunotherapy: expression of membrane complement regulatory proteins (mCRPs) in tumors. Mol Immunol. 2003;40:109–23.
Smoll NR, Drummond KJ. The incidence of medulloblastomas and primitive neurectodermal tumours in adults and children. J Clin Neurosci. 2012;19:1541–4.
McNeil DE, Cote TR, Clegg L, Rorke LB. Incidence and trends in pediatric malignancies medulloblastoma/primitive neuroectodermal tumor: a SEER update. Surveillance Epidemiology and End Results. Med Pediatr Oncol. 2002;39:190–4.
Urberuaga A, Navajas A, Burgos J, Pijoan JI. A review of clinical and histological features of Spanish paediatric medulloblastomas during the last 21 years. Child’s Nerv Syst. 2006;22:466–74.
Verma S, Tavare CJ, Gilles FH. Histologic features and prognosis in pediatric medulloblastoma. Pediatr Dev Pathol. 2008;11:337–43.
Eberhart CG, Kepner JL, Goldthwaite PT, Kun LE, Duffner PK, Friedman HS, et al. Histopathologic grading of medulloblastomas: a Pediatric Oncology Group study. Cancer. 2002;94:552–60.
Kopelson G, Linggood RM, Kleinman GM. Medulloblastoma. The identification of prognostic subgroups and implications for multimodality management. Cancer. 1983;51:312–9.
Caputy AJ, McCullough DC, Manz HJ, Patterson K, Hammock MK. A review of the factors influencing the prognosis of medulloblastoma. The importance of cell differentiation. J Neurosurg. 1987;66:80–7.
van Beek J. Complement activation: beneficial and detrimental effects in the CNS. Ernst Schering Research Foundation workshop 2004; 67–85.
van Beek J, Elward K, Gasque P. Activation of complement in the central nervous system: roles in neurodegeneration and neuroprotection. Ann N Y Acad Sci. 2003;992:56–71.
Markiewski MM, Mastellos D, Tudoran R, DeAngelis RA, Strey CW, Franchini S, et al. C3a and C3b activation products of the third component of complement (C3) are critical for normal liver recovery after toxic injury. J Immunol. 2004;173:747–54 (Baltimore Md: 1950).
Markiewski MM, DeAngelis RA, Benencia F, Ricklin-Lichtsteiner SK, Koutoulaki A, Gerard C, et al. Modulation of the antitumor immune response by complement. Nat Immunol. 2008;9:1225–35.
Nozaki M, Raisler BJ, Sakurai E, Sarma JV, Barnum SR, Lambris JD, et al. Drusen complement components C3a and C5a promote choroidal neovascularization. Proc Natl Acad Sci USA. 2006;103:2328–33.
Nataf S, Stahel PF, Davoust N, Barnum SR. Complement anaphylatoxin receptors on neurons: new tricks for old receptors? Trends Neurosci. 1999;22:397–402.
Ames RS, Li Y, Sarau HM, Nuthulaganti P, Foley JJ, Ellis C, et al. Molecular cloning and characterization of the human anaphylatoxin C3a receptor. J Biol Chem. 1996;271:20231–4.
Acknowledgments
Phillip Bonney was supported by an Oklahoma Shared Clinical and Translational Resources Summer Scholars Program Grant. This study was partially supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20 GM103639 for the use of Histology and Immunohistochemistry Core, which provided staining, immunohistochemistry, and photographic services. We would like to acknowledge Megan Lerner, of the University of Oklahoma, Department of Surgery, for her assistance with immunohistochemical staining. We thank the Peggy and Charles Stephenson Cancer Center at the University of Oklahoma, Oklahoma City, OK and an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20 GM103639 for the use of Histology and Immunohistochemistry Core, which provided staining, immunohistochemistry, and photographic services.
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The authors declare that they have no conflict of interest.
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Maurer, A.J., Bonney, P.A., Toho, L.C. et al. Tumor necrosis-initiated complement activation stimulates proliferation of medulloblastoma cells. Inflamm. Res. 64, 185–192 (2015). https://doi.org/10.1007/s00011-015-0796-y
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DOI: https://doi.org/10.1007/s00011-015-0796-y