PLEKHG5 regulates autophagy, survival and MGMT expression in U251-MG glioblastoma cells

Abstract A signalling pathway involving PLEKHG5 (guanine exchange factor) for the Ras superfamily member RAB26 to transcription factor NF-κB was discovered in autophagy. PLEKHG5 was reported in glioblastoma multiforme (GBM) and correlates with patient survival. Thus, the generation of a cellular mod...

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Autores principales:	Kaya Elisa Witte, Carsten Slotta, Melanie Lütkemeyer, Angelika Kitke, Roland Coras, Matthias Simon, Christian Kaltschmidt, Barbara Kaltschmidt
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9ca64becd4b34645835d2d046c52bd05
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spelling	oai:doaj.org-article:9ca64becd4b34645835d2d046c52bd052021-12-02T12:42:18ZPLEKHG5 regulates autophagy, survival and MGMT expression in U251-MG glioblastoma cells10.1038/s41598-020-77958-32045-2322https://doaj.org/article/9ca64becd4b34645835d2d046c52bd052020-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-77958-3https://doaj.org/toc/2045-2322Abstract A signalling pathway involving PLEKHG5 (guanine exchange factor) for the Ras superfamily member RAB26 to transcription factor NF-κB was discovered in autophagy. PLEKHG5 was reported in glioblastoma multiforme (GBM) and correlates with patient survival. Thus, the generation of a cellular model for understanding PLEKHG5 signalling is the study purpose. We generated a CRISPR/Cas9-mediated knockout of PLEKHG5 in U251-MG glioblastoma cells and analysed resulting changes. Next, we used a mRFP-GFP-LC3+ reporter for visualisation of autophagic defects and rescued the phenotype of PLEKHG5 wildtype via transduction of a constitutively active RAB26QL-plasmid. Effects of overexpressing RAB26 were investigated and correlated with the O6-methylguanine-DNA methyltransferase (MGMT) and cellular survival. PLEKHG5 knockout showed changes in morphology, loss of filopodia and higher population doubling times. Accumulation of autolysosomes was resulted by decreased LAMP-1 in PLEKHG5-deficient cells. Rescue of PLEKHG5 −/− restored the downregulation of RhoA activity, showed faster response to tumour necrosis factor and better cellular fitness. MGMT expression was activated after RAB26 overexpression compared to non-transduced cells. Survival of PLEKHG5 knockout was rescued together with sensitivity to temozolomide by RAB26QL. This study provides new insights in the PLEKHG5/RAB26 signalling within U251-MG cells, which suggests potential therapeutic strategies in other glioma cells and further in primary GBM.Kaya Elisa WitteCarsten SlottaMelanie LütkemeyerAngelika KitkeRoland CorasMatthias SimonChristian KaltschmidtBarbara KaltschmidtNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-19 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Kaya Elisa Witte Carsten Slotta Melanie Lütkemeyer Angelika Kitke Roland Coras Matthias Simon Christian Kaltschmidt Barbara Kaltschmidt PLEKHG5 regulates autophagy, survival and MGMT expression in U251-MG glioblastoma cells
description	Abstract A signalling pathway involving PLEKHG5 (guanine exchange factor) for the Ras superfamily member RAB26 to transcription factor NF-κB was discovered in autophagy. PLEKHG5 was reported in glioblastoma multiforme (GBM) and correlates with patient survival. Thus, the generation of a cellular model for understanding PLEKHG5 signalling is the study purpose. We generated a CRISPR/Cas9-mediated knockout of PLEKHG5 in U251-MG glioblastoma cells and analysed resulting changes. Next, we used a mRFP-GFP-LC3+ reporter for visualisation of autophagic defects and rescued the phenotype of PLEKHG5 wildtype via transduction of a constitutively active RAB26QL-plasmid. Effects of overexpressing RAB26 were investigated and correlated with the O6-methylguanine-DNA methyltransferase (MGMT) and cellular survival. PLEKHG5 knockout showed changes in morphology, loss of filopodia and higher population doubling times. Accumulation of autolysosomes was resulted by decreased LAMP-1 in PLEKHG5-deficient cells. Rescue of PLEKHG5 −/− restored the downregulation of RhoA activity, showed faster response to tumour necrosis factor and better cellular fitness. MGMT expression was activated after RAB26 overexpression compared to non-transduced cells. Survival of PLEKHG5 knockout was rescued together with sensitivity to temozolomide by RAB26QL. This study provides new insights in the PLEKHG5/RAB26 signalling within U251-MG cells, which suggests potential therapeutic strategies in other glioma cells and further in primary GBM.
format	article
author	Kaya Elisa Witte Carsten Slotta Melanie Lütkemeyer Angelika Kitke Roland Coras Matthias Simon Christian Kaltschmidt Barbara Kaltschmidt
author_facet	Kaya Elisa Witte Carsten Slotta Melanie Lütkemeyer Angelika Kitke Roland Coras Matthias Simon Christian Kaltschmidt Barbara Kaltschmidt
author_sort	Kaya Elisa Witte
title	PLEKHG5 regulates autophagy, survival and MGMT expression in U251-MG glioblastoma cells
title_short	PLEKHG5 regulates autophagy, survival and MGMT expression in U251-MG glioblastoma cells
title_full	PLEKHG5 regulates autophagy, survival and MGMT expression in U251-MG glioblastoma cells
title_fullStr	PLEKHG5 regulates autophagy, survival and MGMT expression in U251-MG glioblastoma cells
title_full_unstemmed	PLEKHG5 regulates autophagy, survival and MGMT expression in U251-MG glioblastoma cells
title_sort	plekhg5 regulates autophagy, survival and mgmt expression in u251-mg glioblastoma cells
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/9ca64becd4b34645835d2d046c52bd05
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_version_	1718393700977999872

PLEKHG5 regulates autophagy, survival and MGMT expression in U251-MG glioblastoma cells

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