Clustered localization of EGFRvIII in glioblastoma cells as detected by high precision localization microscopy

• Verfasst von: Boyd, Philip S. [VerfasserIn]
• Verfasst von: Struve, Nina [VerfasserIn]
• Verfasst von: Bach, Margund [VerfasserIn]
• Verfasst von: Eberle, Jan Philipp [VerfasserIn]
• Verfasst von: Gote, Martin [VerfasserIn]
• Verfasst von: Schock, Florian [VerfasserIn]
• Verfasst von: Cremer, Christoph [VerfasserIn]
• Verfasst von: Kriegs, Malte [VerfasserIn]
• Verfasst von: Hausmann, Michael [VerfasserIn]
• Titel: Clustered localization of EGFRvIII in glioblastoma cells as detected by high precision localization microscopy
• Verf.angabe: Philip S. Boyd, Nina Struve, Margund Bach, Jan Philipp Eberle, Martin Gote, Florian Schock, Christoph Cremer, Malte Kriegs, Michael Hausmann
• E-Jahr: 2016
• Jahr: 09 Nov 2016
• Umfang: 11 S.
• Fussnoten: Gesehen am 18.05.2020
• Titel Quelle: Enthalten in: Nanoscale
• Ort Quelle: Cambridge : RSC Publ., 2009
• Jahr Quelle: 2016
• Band/Heft Quelle: 8(2016), 48, Seite 20037-20047
• ISSN Quelle: 2040-3372
• DOI: doi:10.1039/C6NR05880A
• Datenträger: Online-Ressource
• Sprache: eng
• K10plus-PPN: 1698455046

Abstract

For receptor tyrosine kinases supramolecular organization on the cell membrane is critical for their function. Super-resolution fluorescence microscopy techniques have offered new opportunities for the analysis of single receptor localization. Here, we analysed the cluster formation of the epidermal growth factor receptor variant III (EGFRvIII), a deletion variant which is expressed in glioblastoma. The constitutively activated variant EGFRvIII is expressed in cells with an egfr gene amplification and is thought to enhance the tumorigenic potential especially of glioblastoma cells. Due to the lack of an adequate model system, it is still unclear how endogenous EGFRvIII expression alters cellular signalling and if it is organized in clusters like the wild type receptor. We have recently described the establishment of two pairs of iso-genetic cell lines (BS153 and DKMG), displaying endogenous EGFRvIII expression or not. Using these cell lines we investigated single receptor localization of EGFRvIII by high precision localization microscopy. Cluster analysis revealed that EGFRvIII is present in clusters on the surface of the cells, with about 60% or even more receptor molecules being assembled in clusters of approximately 100 nm in diameter whereby the cluster definition was iteratively determined. The signal to signal distance may indicate dimer formation while signal quantification indicates 1 × 106-5 × 106 EGFRvIII molecules per cell. Altogether, these data give unique insights into the membrane surface localization of EGFRvIII in glioblastoma cells. These insights will help to unveil the function of this tumour associated receptor variant which might lead to a better understanding of glioblastoma and therefore could lead to improved therapy approaches.