Versatile Role of Rab27a in Glioma: Effects on Release of Extracellular Vesicles, Cell Viability, and Tumor Progression.

Autor:	van Solinge TS; Department of Neurology and Radiology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States.; NeuroDiscovery Center, Harvard Medical School, Boston, MA, United States.; Department of Neurosurgery, Leiden University Medical Center, Leiden, Netherlands., Abels ER; Department of Neurology and Radiology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States.; NeuroDiscovery Center, Harvard Medical School, Boston, MA, United States., van de Haar LL; Department of Neurology and Radiology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States.; NeuroDiscovery Center, Harvard Medical School, Boston, MA, United States., Hanlon KS; Department of Neurobiology, Harvard Medical School, Boston, MA, United States.; Molecular Neurogenetics Unit, Department of Neurology, Massachusetts General Hospital, Charlestown, MA, United States., Maas SLN; Department of Neurosurgery, UMC Utrecht Brain Center, Utrecht University, Utrecht, Netherlands.; Department of Pathology, University Medical Center Utrecht, Utrecht University, Utrecht, Netherlands., Schnoor R; Department of Neurosurgery, UMC Utrecht Brain Center, Utrecht University, Utrecht, Netherlands., de Vrij J; Department of Neurosurgery, Brain Tumor Center, Erasmus Medical Center, Rotterdam, Netherlands., Breakefield XO; Department of Neurology and Radiology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States.; NeuroDiscovery Center, Harvard Medical School, Boston, MA, United States., Broekman MLD; Department of Neurology and Radiology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States.; NeuroDiscovery Center, Harvard Medical School, Boston, MA, United States.; Department of Neurosurgery, Leiden University Medical Center, Leiden, Netherlands.; Department of Neurosurgery, Haaglanden Medical Center, The Hague, Netherlands.
Jazyk:	angličtina
Zdroj:	Frontiers in molecular biosciences [Front Mol Biosci] 2020 Nov 12; Vol. 7, pp. 554649. Date of Electronic Publication: 2020 Nov 12 (Print Publication: 2020).
DOI:	10.3389/fmolb.2020.554649
Abstrakt:	Introduction: Glioma cells exert influence over the tumor-microenvironment in part through the release of extracellular vesicles (EVs), membrane-enclosed structures containing proteins, lipids, and RNAs. In this study, we evaluated the function of Ras-associated protein 27a (Rab27a) in glioma and evaluated the feasibility of assessing its role in EV release in glioma cells in vitro and in vivo . Methods: Rab27a was knocked down via a short hairpin RNA (shRNA) stably expressed in mouse glioma cell line GL261, with a scrambled shRNA as control. EVs were isolated by ultracentrifugation and quantified with Nanoparticle Tracking Analysis (NTA) and Tunable Resistive Pulse Sensing (TRPS). CellTiter-Glo viability assays and cytokine arrays were used to evaluate the impact of Rab27a knockdown. GL261.shRab27a cells and GL261.shControl were implanted into the left striatum of eight mice to assess tumor growth and changes in the tumor microenvironment. Results: Knockdown of Rab27a in GL261 glioma cells decreased the release of small EVs isolated at 100,000 × g in vitro ( p = 0.005), but not the release of larger EVs, isolated at 10,000 × g . GL261.shRab27a cells were less viable compared to the scramble control in vitro ( p < 0.005). A significant increase in CCL2 expression in shRab27a GL261 cells was also observed ( p < 0.001). However, in vivo there was no difference in tumor growth or overall survival between the two groups, while shRab27a tumors showed lower proliferation at the tumor borders. Decreased infiltration of IBA1 positive macrophages and microglia, but not FoxP3 positive regulatory T cells was observed. Conclusion: Rab27a plays an important role in the release of small EVs from glioma cells, and also in their viability and expression of CCL2 in vitro. As interference in Rab27a expression influences glioma cell viability and expression profiles, future studies should be cautious in using the knockdown of Rab27a as a means of studying the role of small EVs in glioma growth. (Copyright © 2020 van Solinge, Abels, van de Haar, Hanlon, Maas, Schnoor, de Vrij, Breakefield and Broekman.)
Databáze:	MEDLINE
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