P12.04.A Exosomes from glioma associated sphere forming cells induce a transition of invasive phenotype via transfer of EMP2 and CA9

Abstract Background Glioblastoma multiforme (GBM) mostly occurs local recurrence at normal parenchyme adjacent tumor despite of conventional treatment. Glioma stem like cells (GSC) forming intratumoral heterogeneity within the GBM acquired microenvironmental adaptation by inter-exosomal contents exchange between heterogenic cells. In addition, GSC has an invasive potential as human GBM. Therefore, we investigate whether exosomal proteins affect tissue invasion in Material and Methods Exosomes were isolated Size-Exclusion method from conditioned media validated Electron microscope Immunoblot assay. Exosomal proteomics examined with Liquid Chromatography-Mass Spectrometry (LC/MS). To produce fluorescent exosome, bi-cistron vectors cloned shRNA CD63-GFP. identify effect tranfected exosomes treated to recipient 3D assay mouse intracranial model. Results Firstly, dichotomized two groups following matrigel derived orthotopic CSC2 X01 GSCs revealed highly phenotype whereas 83NS 528NS did not. Exosome was each group identified CD63 expression or electron microscopy. analysis, hypoxia, extracellular matrix organization, GTPase cycle related enriched cell’s exosome. Among them, focused carbonic anhydrase IX (CA9) epithelial membrane protein 2 (EMP2) on its permissive role glioblastoma respectively. CA9 EMP2 mRNA levels verified their high detected compared low one GSCs. evaluate effects exosome mediated potential, viral composed target gene knockdown fluorescence used detect intracellular transfer. Interestingly, decreased phosphorylated FAK, a key marker, observed after Lentiviral CA9- EMP2-knockdown CSC2. are effector responsible for glioma invasion, donor (Exo-CSC2-sh-CA9 Exo-CSC2-EMP2, transfection CSC2s) non assay, Exo-CSC2-shCA9 Exo-CSC2-shEMP2 significantly Exo-CSC2-shEV. These features found model well. Conclusion Together these, conclude that induces transition via transfer proteins.