Identification of transcriptional targets of GSK3 involved in glioblastoma invasion

Anke Bruning-Richardson, Alastair Droop, Daniel Tams, Marjorie Boissinot, Josie Hayes, Vinton Cheng, Julia V. Cockle, Azzam Ismail, Ruth Morton, Filomena Esteves, Michel Mittelbronn, Sean Lawler, Susan C Short, Georgia Mavria

Research output: Contribution to journalMeeting Abstract

Abstract

INTRODUCTION
Glioblastoma multiforme remains one of the most difficult to treat cancers due to the highly infiltrative nature of cancer cells. Targeting the invasive properties of glioblastoma presents a novel treatment avenue. Recently, specific GSK-3 beta (GSK3b) inhibitors such as Bio-Indirubin (BIO), have been described that inhibit the migratory activity of glioma cells. In this study, the GSK-3b downstream targets were investigated.

METHOD
Genome-wide transcriptional profiling was used in order to identify GSK3b-driven target genes, and a transcriptional signature for glioblastoma invasion. U251 cells and patient-derived GBM1 cells were treated with the GSK3b inhibitor BIO. Microarray data obtained was mined for deregulated genes with known migratory roles. The up-regulation or down-regulation of candidate genes was confirmed by Western blotting and immunofluorescence. Knockdown of two candidate genes by RNAi, ARHGAP12 and ARHGAP29 was performed, and the effects on invasive properties and characteristics were assessed by 3D spheroid assays, live cell imaging and immunofluorescence, in order to investigate the effects of knockdowns on invasion.

RESULTS
We have identified an invasion signature downstream of GSK3b, which includes activators and inactivators of the Rho family of small GTPases. Knockdown of the Rac1 inactivator ARHGAP12 increased migration, whereas knockdown of the RhoA inactivator ARHGAP29 decreased migration as assessed by live cell imaging and 3D spheroid invasion assays.

CONCLUSIONS
This study, which for the first time investigates downstream targets of GSK-3b in glioblastoma, has identified an invasion signature which includes regulators of the Rho family of small GTPases. Identified genes will be potential candidates for genetic involvement in glioblastoma progression, and candidate diagnostic and prognostic biomarkers. Candidates will be assessed as potential therapeutic targets.
LanguageEnglish
Pages26
Number of pages1
JournalNeuro-Oncology
Volume20
Issue numberS1
DOIs
Publication statusPublished - Jan 2018
Externally publishedYes
EventBritish Neuro Oncology Society Conference: Enhancing Science, Enhancing Survival - Edinburgh, United Kingdom
Duration: 21 Jun 201723 Jun 2017
https://www.bnos.org.uk/wp-content/uploads/2017/08/BNOS-2017-Conference-Report.pdf (Link to Conference Report)

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Glioblastoma
Glycogen Synthase Kinase 3
Monomeric GTP-Binding Proteins
Genes
Fluorescent Antibody Technique
RNA Interference
Glioma
Neoplasms
Up-Regulation
Down-Regulation
Biomarkers
Western Blotting
Therapeutics

Cite this

Bruning-Richardson, Anke ; Droop, Alastair ; Tams, Daniel ; Boissinot, Marjorie ; Hayes, Josie ; Cheng, Vinton ; Cockle, Julia V. ; Ismail, Azzam ; Morton, Ruth ; Esteves, Filomena ; Mittelbronn, Michel ; Lawler, Sean ; Short, Susan C ; Mavria, Georgia. / Identification of transcriptional targets of GSK3 involved in glioblastoma invasion. In: Neuro-Oncology. 2018 ; Vol. 20, No. S1. pp. 26.
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title = "Identification of transcriptional targets of GSK3 involved in glioblastoma invasion",
abstract = "INTRODUCTIONGlioblastoma multiforme remains one of the most difficult to treat cancers due to the highly infiltrative nature of cancer cells. Targeting the invasive properties of glioblastoma presents a novel treatment avenue. Recently, specific GSK-3 beta (GSK3b) inhibitors such as Bio-Indirubin (BIO), have been described that inhibit the migratory activity of glioma cells. In this study, the GSK-3b downstream targets were investigated.METHODGenome-wide transcriptional profiling was used in order to identify GSK3b-driven target genes, and a transcriptional signature for glioblastoma invasion. U251 cells and patient-derived GBM1 cells were treated with the GSK3b inhibitor BIO. Microarray data obtained was mined for deregulated genes with known migratory roles. The up-regulation or down-regulation of candidate genes was confirmed by Western blotting and immunofluorescence. Knockdown of two candidate genes by RNAi, ARHGAP12 and ARHGAP29 was performed, and the effects on invasive properties and characteristics were assessed by 3D spheroid assays, live cell imaging and immunofluorescence, in order to investigate the effects of knockdowns on invasion.RESULTSWe have identified an invasion signature downstream of GSK3b, which includes activators and inactivators of the Rho family of small GTPases. Knockdown of the Rac1 inactivator ARHGAP12 increased migration, whereas knockdown of the RhoA inactivator ARHGAP29 decreased migration as assessed by live cell imaging and 3D spheroid invasion assays.CONCLUSIONSThis study, which for the first time investigates downstream targets of GSK-3b in glioblastoma, has identified an invasion signature which includes regulators of the Rho family of small GTPases. Identified genes will be potential candidates for genetic involvement in glioblastoma progression, and candidate diagnostic and prognostic biomarkers. Candidates will be assessed as potential therapeutic targets.",
author = "Anke Bruning-Richardson and Alastair Droop and Daniel Tams and Marjorie Boissinot and Josie Hayes and Vinton Cheng and Cockle, {Julia V.} and Azzam Ismail and Ruth Morton and Filomena Esteves and Michel Mittelbronn and Sean Lawler and Short, {Susan C} and Georgia Mavria",
year = "2018",
month = "1",
doi = "10.1093/neuonc/nox238.117",
language = "English",
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Bruning-Richardson, A, Droop, A, Tams, D, Boissinot, M, Hayes, J, Cheng, V, Cockle, JV, Ismail, A, Morton, R, Esteves, F, Mittelbronn, M, Lawler, S, Short, SC & Mavria, G 2018, 'Identification of transcriptional targets of GSK3 involved in glioblastoma invasion', Neuro-Oncology, vol. 20, no. S1, pp. 26. https://doi.org/10.1093/neuonc/nox238.117

Identification of transcriptional targets of GSK3 involved in glioblastoma invasion. / Bruning-Richardson, Anke; Droop, Alastair; Tams, Daniel; Boissinot, Marjorie; Hayes, Josie; Cheng, Vinton; Cockle, Julia V.; Ismail, Azzam; Morton, Ruth; Esteves, Filomena; Mittelbronn, Michel; Lawler, Sean; Short, Susan C; Mavria, Georgia.

In: Neuro-Oncology, Vol. 20, No. S1, 01.2018, p. 26.

Research output: Contribution to journalMeeting Abstract

TY - JOUR

T1 - Identification of transcriptional targets of GSK3 involved in glioblastoma invasion

AU - Bruning-Richardson, Anke

AU - Droop, Alastair

AU - Tams, Daniel

AU - Boissinot, Marjorie

AU - Hayes, Josie

AU - Cheng, Vinton

AU - Cockle, Julia V.

AU - Ismail, Azzam

AU - Morton, Ruth

AU - Esteves, Filomena

AU - Mittelbronn, Michel

AU - Lawler, Sean

AU - Short, Susan C

AU - Mavria, Georgia

PY - 2018/1

Y1 - 2018/1

N2 - INTRODUCTIONGlioblastoma multiforme remains one of the most difficult to treat cancers due to the highly infiltrative nature of cancer cells. Targeting the invasive properties of glioblastoma presents a novel treatment avenue. Recently, specific GSK-3 beta (GSK3b) inhibitors such as Bio-Indirubin (BIO), have been described that inhibit the migratory activity of glioma cells. In this study, the GSK-3b downstream targets were investigated.METHODGenome-wide transcriptional profiling was used in order to identify GSK3b-driven target genes, and a transcriptional signature for glioblastoma invasion. U251 cells and patient-derived GBM1 cells were treated with the GSK3b inhibitor BIO. Microarray data obtained was mined for deregulated genes with known migratory roles. The up-regulation or down-regulation of candidate genes was confirmed by Western blotting and immunofluorescence. Knockdown of two candidate genes by RNAi, ARHGAP12 and ARHGAP29 was performed, and the effects on invasive properties and characteristics were assessed by 3D spheroid assays, live cell imaging and immunofluorescence, in order to investigate the effects of knockdowns on invasion.RESULTSWe have identified an invasion signature downstream of GSK3b, which includes activators and inactivators of the Rho family of small GTPases. Knockdown of the Rac1 inactivator ARHGAP12 increased migration, whereas knockdown of the RhoA inactivator ARHGAP29 decreased migration as assessed by live cell imaging and 3D spheroid invasion assays.CONCLUSIONSThis study, which for the first time investigates downstream targets of GSK-3b in glioblastoma, has identified an invasion signature which includes regulators of the Rho family of small GTPases. Identified genes will be potential candidates for genetic involvement in glioblastoma progression, and candidate diagnostic and prognostic biomarkers. Candidates will be assessed as potential therapeutic targets.

AB - INTRODUCTIONGlioblastoma multiforme remains one of the most difficult to treat cancers due to the highly infiltrative nature of cancer cells. Targeting the invasive properties of glioblastoma presents a novel treatment avenue. Recently, specific GSK-3 beta (GSK3b) inhibitors such as Bio-Indirubin (BIO), have been described that inhibit the migratory activity of glioma cells. In this study, the GSK-3b downstream targets were investigated.METHODGenome-wide transcriptional profiling was used in order to identify GSK3b-driven target genes, and a transcriptional signature for glioblastoma invasion. U251 cells and patient-derived GBM1 cells were treated with the GSK3b inhibitor BIO. Microarray data obtained was mined for deregulated genes with known migratory roles. The up-regulation or down-regulation of candidate genes was confirmed by Western blotting and immunofluorescence. Knockdown of two candidate genes by RNAi, ARHGAP12 and ARHGAP29 was performed, and the effects on invasive properties and characteristics were assessed by 3D spheroid assays, live cell imaging and immunofluorescence, in order to investigate the effects of knockdowns on invasion.RESULTSWe have identified an invasion signature downstream of GSK3b, which includes activators and inactivators of the Rho family of small GTPases. Knockdown of the Rac1 inactivator ARHGAP12 increased migration, whereas knockdown of the RhoA inactivator ARHGAP29 decreased migration as assessed by live cell imaging and 3D spheroid invasion assays.CONCLUSIONSThis study, which for the first time investigates downstream targets of GSK-3b in glioblastoma, has identified an invasion signature which includes regulators of the Rho family of small GTPases. Identified genes will be potential candidates for genetic involvement in glioblastoma progression, and candidate diagnostic and prognostic biomarkers. Candidates will be assessed as potential therapeutic targets.

U2 - 10.1093/neuonc/nox238.117

DO - 10.1093/neuonc/nox238.117

M3 - Meeting Abstract

VL - 20

SP - 26

JO - Neuro-Oncology

T2 - Neuro-Oncology

JF - Neuro-Oncology

SN - 1522-8517

IS - S1

ER -