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Learn more: of α2/β1 integrin upregulates functional glioma-derived stem cells.
Multipotent glioma-derived stem cells (gSCs) differentiate into the fate of most glioma cells and have tumorigenic activity. However, the mechanism regulating the maintenance of the undifferentiated state of gSCs is unknown. Here, we found that glioma cells generated from patient-derived tumor tissue formed spherical clusters on Matrigel, which were enriched for the glioma-associated markers CD133 and Nestin. To define the molecular changes associated with cluster formation, we performed comparative microarray analysis of cluster and single cell samples by RNA-sequencing. Single cells formed compact spheroids by cell aggregation. In contrast, cluster cells formed floating spheroids composed of multiple cell layers. Of the 1,054 genes upregulated in clusters, more than 50% were related to the maintenance and development of neural stem cells. We found that cluster formation was blocked by R5 peptide, which was derived from neural cell adhesion molecule and binds to α2/β1 integrins. R5 peptide binds to a unique receptor expressed on gSCs, the caveolin-enriched membrane fraction. Treatment of gSCs with R5 peptide reduced cell aggregation in serum-containing medium, whereas R5 peptide did not affect the stemness of gSCs cultured in neurosphere medium. R5 peptide reduced the formation of floating, multilayered spheroids. In conclusion, the down-regulation of α2/β1 integrin by R5 peptide may contribute to cluster formation and to the maintenance of the undifferentiated phenotype of gSCs in vivo.Thank you
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