GDNF mediates glioblastoma-induced microglia attraction but not astrogliosis
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High-grade gliomas are the most common primary brain tumors. Their malignancy is promoted by the complex crosstalk between different cell types in the central nervous system. Microglia/brain macrophages infiltrate high-grade gliomas and contribute to their progression. To identify factors that mediate the attraction of microglia/macrophages to malignant brain tumors, we established a glioma cell encapsulation model that was applied in vivo. Mouse GL261 glioma cell line and human high-grade glioma cells were seeded into hollow fibers (HF) that allow the passage of soluble molecules but not cells. The glioma cell containing HF were implanted into one brain hemisphere and simultaneously HF with non-transformed fibroblasts (controls) were introduced into the contralateral hemisphere. Implanted mouse and human glioma- but not fibroblast-containing HF attracted microglia and up-regulated immunoreactivity for GFAP, which is a marker of astrogliosis. In this study, we identified GDNF as an important factor for microglial attraction: (1) GL261 and human glioma cells secret GDNF, (2) reduced GDNF production by siRNA in GL261 in mouse glioma cells diminished attraction of microglia, (3) over-expression of GDNF in fibroblasts promoted microglia attraction in our HF assay. In vitro migration assays also showed that GDNF is a strong chemoattractant for microglia. While GDNF release from human or mouse glioma had a profound effect on microglial attraction, the glioma-induced astrogliosis was not affected. Finally, we could show that injection of GL261 mouse glioma cells with GDNF knockdown by shRNA into mouse brains resulted in reduced tumor expansion and improved survival as compared to injection of control cells.
KeywordsGlioblastoma Microglia GDNF Astrocyte
This work was supported by the graduate school of NeuroCure at the Charité, Berlin, (stipend to M. C. Ku) and by Deutsche Forschungsgemeinschaft (TR 43). We are thankful to Prof. Carlos Ibanez for providing GDNF cDNA construct for transfection and Prof. Jochen Meier for providing human brain tissue. We thank Dr. Zoltan Cseresnyes and Dr. Anje Sporbert for technical assistance with confocal microscopy. We appreciate the support of Babette Dieringer for MR imaging and Maria Pannell for manuscript proof reading.
Conflict of interest
The authors declare that they have no conflict of interest.
- 2.Badie B, Schartner J, Klaver J, Vorpahl J (1999) In vitro modulation of microglia motility by glioma cells is mediated by hepatocyte growth factor/scatter factor. Neurosurgery 44(5):1077–1082 (discussion 1082–1073)Google Scholar
- 6.Dhandapani KM, Khan MM, Wade FM, Wakade C, Mahesh VB, Brann DW (2007) Induction of transforming growth factor-beta1 by basic fibroblast growth factor in rat C6 glioma cells and astrocytes is mediated by MEK/ERK signaling and AP-1 activation. J Neurosci Res 85(5):1033–1045. doi: 10.1002/jnr.21182 PubMedCrossRefGoogle Scholar
- 8.Edwards LA, Woolard K, Son MJ, Li A, Lee J, Ene C, Mantey SA, Maric D, Song H, Belova G, Jensen RT, Zhang W, Fine HA (2011) Effect of brain- and tumor-derived connective tissue growth factor on glioma invasion. J Natl Cancer Inst 103(15):1162–1178. doi: djr22410.1093/jnci/djr224 PubMedCrossRefGoogle Scholar
- 9.Franklin K, Paxinos G (2007) The mouse brain in stereotaxic coordinates, 3rd edn. Academic Press, San DiegoGoogle Scholar
- 11.Glass R, Synowitz M, Kronenberg G, Walzlein JH, Markovic DS, Wang LP, Gast D, Kiwit J, Kempermann G, Kettenmann H (2005) Glioblastoma-induced attraction of endogenous neural precursor cells is associated with improved survival. J Neurosci 25(10):2637–2646. doi: 25/10/263710.1523/JNEUROSCI.5118-04.2005 PubMedCrossRefGoogle Scholar
- 12.Han Q, Sun W, Lin H, Zhao W, Gao Y, Zhao Y, Chen B, Xiao Z, Hu W, Li Y, Yang B, Dai J (2009) Linear ordered collagen scaffolds loaded with collagen-binding brain-derived neurotrophic factor improve the recovery of spinal cord injury in rats. Tissue Eng Part A 15(10):2927–2935. doi: 10.1089/ten.TEA.2008.0506 PubMedCrossRefGoogle Scholar
- 14.Kenig S, Alonso MB, Mueller MM, Lah TT (2009) Glioblastoma and endothelial cells cross-talk, mediated by SDF-1, enhances tumour invasion and endothelial proliferation by increasing expression of cathepsins B, S, and MMP-9. Cancer Lett 289(1):53–61. doi: 10.1016/j.canlet.2009.07.014 PubMedCrossRefGoogle Scholar
- 19.Lang AE, Gill S, Patel NK, Lozano A, Nutt JG, Penn R, Brooks DJ, Hotton G, Moro E, Heywood P, Brodsky MA, Burchiel K, Kelly P, Dalvi A, Scott B, Stacy M, Turner D, Wooten VG, Elias WJ, Laws ER, Dhawan V, Stoessl AJ, Matcham J, Coffey RJ, Traub M (2006) Randomized controlled trial of intraputamenal glial cell line-derived neurotrophic factor infusion in Parkinson disease. Ann Neurol 59(3):459–466. doi: 10.1002/ana.20737 PubMedCrossRefGoogle Scholar
- 20.Le DM, Besson A, Fogg DK, Choi KS, Waisman DM, Goodyer CG, Rewcastle B, Yong VW (2003) Exploitation of astrocytes by glioma cells to facilitate invasiveness: a mechanism involving matrix metalloproteinase-2 and the urokinase-type plasminogen activator-plasmin cascade. J Neurosci 23(10):4034–4043PubMedGoogle Scholar
- 26.Markovic DS, Vinnakota K, Chirasani S, Synowitz M, Raguet H, Stock K, Sliwa M, Lehmann S, Kalin R, van Rooijen N, Holmbeck K, Heppner FL, Kiwit J, Matyash V, Lehnardt S, Kaminska B, Glass R, Kettenmann H (2009) Gliomas induce and exploit microglial MT1-MMP expression for tumor expansion. Proc Natl Acad Sci USA 106(30):12530–12535. doi: 10.1073/pnas.0804273106 PubMedCrossRefGoogle Scholar