Finding the brake: dissecting the RhoA pathway regulating glioma cell migration

Sophie Taylor, Sabine Knipp, Georgia Mavria, John Ladbury, Anke Bruning-Richardson

Research output: Contribution to journalMeeting Abstract

Abstract

Cell migration is one of the hallmarks of cancer. Cancer cells can adopt two main migratory strategies displaying either a mesenchymal or amoeboid phenotype. Targeting cell migration presents an opportunity in improving treatment of invasive and migratory tumours, however the cellular mechanisms that control the cell migration phenotypes in high-grade gliomas, such as Glioblastoma multiforme, are still not fully understood. In previous work we used 2D and 3D migration assays to screen a panel of inhibitors targeting the actin polymerisation pathway, a key regulator of cell migration in glioma. Amongst RhoA pathway inhibitors, which are key negative regulators of migration, we have identified one compound, CCG-1423, that uniquely failed to inhibit migration. CCG-1423 induces a reduced expression of the adhesion protein CCN1 by blocking nuclear import of MKL-1, a transcriptional co-activator. We suggest that CCG-1423 induces a mesenchymal transition, which permits continued migration. To confirm the effect of CCG-1423 on CCN1 levels we used ELISA to detect secreted CCN1 levels in culture supernatants in treated and non-treated U251 and U87 cells. CCG-1423 significantly reduced CCN1 levels in supernatant of both cell lines compared to untreated controls. Further biochemical analysis was performed to establish levels of MKL-1 in treated versus untreated cells and to elucidate its association with other proteins. Preliminary data indicate an increase in MKL-1 concentration in treated cells, consistent with a build-up of MKL-1 in the cytosol due to lack of nuclear import, as previously reported. Further detailed characterisation of the RhoA signalling pathway in glioma will provide an understanding of the role of MKL-1 in mesenchymal migration and the effect of CCG-1423 on mesenchymal switching. We will next characterise the interactions between CCG-1423 and MKL-1, and use pharmacological and/or genetic manipulation to refine combination treatments to target both mesenchymal and amoeboid cell migration.
LanguageEnglish
Pages21-22
Number of pages2
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)

Fingerprint

Glioma
Cell Movement
Cell Nucleus Active Transport
Cysteine-Rich Protein 61
Phenotype
Neoplasms
Glioblastoma
CCG 1423
Polymerization
Cytosol
Actins
Enzyme-Linked Immunosorbent Assay
Pharmacology
Cell Line
Therapeutics
Proteins

Cite this

Taylor, Sophie ; Knipp, Sabine ; Mavria, Georgia ; Ladbury, John ; Bruning-Richardson, Anke. / Finding the brake : dissecting the RhoA pathway regulating glioma cell migration. In: Neuro-Oncology. 2018 ; Vol. 20, No. S1. pp. 21-22.
@article{e3d316ac0fe84d91bbd651719e318e1f,
title = "Finding the brake: dissecting the RhoA pathway regulating glioma cell migration",
abstract = "Cell migration is one of the hallmarks of cancer. Cancer cells can adopt two main migratory strategies displaying either a mesenchymal or amoeboid phenotype. Targeting cell migration presents an opportunity in improving treatment of invasive and migratory tumours, however the cellular mechanisms that control the cell migration phenotypes in high-grade gliomas, such as Glioblastoma multiforme, are still not fully understood. In previous work we used 2D and 3D migration assays to screen a panel of inhibitors targeting the actin polymerisation pathway, a key regulator of cell migration in glioma. Amongst RhoA pathway inhibitors, which are key negative regulators of migration, we have identified one compound, CCG-1423, that uniquely failed to inhibit migration. CCG-1423 induces a reduced expression of the adhesion protein CCN1 by blocking nuclear import of MKL-1, a transcriptional co-activator. We suggest that CCG-1423 induces a mesenchymal transition, which permits continued migration. To confirm the effect of CCG-1423 on CCN1 levels we used ELISA to detect secreted CCN1 levels in culture supernatants in treated and non-treated U251 and U87 cells. CCG-1423 significantly reduced CCN1 levels in supernatant of both cell lines compared to untreated controls. Further biochemical analysis was performed to establish levels of MKL-1 in treated versus untreated cells and to elucidate its association with other proteins. Preliminary data indicate an increase in MKL-1 concentration in treated cells, consistent with a build-up of MKL-1 in the cytosol due to lack of nuclear import, as previously reported. Further detailed characterisation of the RhoA signalling pathway in glioma will provide an understanding of the role of MKL-1 in mesenchymal migration and the effect of CCG-1423 on mesenchymal switching. We will next characterise the interactions between CCG-1423 and MKL-1, and use pharmacological and/or genetic manipulation to refine combination treatments to target both mesenchymal and amoeboid cell migration.",
author = "Sophie Taylor and Sabine Knipp and Georgia Mavria and John Ladbury and Anke Bruning-Richardson",
year = "2018",
month = "1",
doi = "10.1093/neuonc/nox238.095",
language = "English",
volume = "20",
pages = "21--22",
journal = "Neuro-Oncology",
issn = "1522-8517",
publisher = "Oxford University Press",
number = "S1",

}

Finding the brake : dissecting the RhoA pathway regulating glioma cell migration. / Taylor, Sophie; Knipp, Sabine; Mavria, Georgia; Ladbury, John; Bruning-Richardson, Anke.

In: Neuro-Oncology, Vol. 20, No. S1, 01.2018, p. 21-22.

Research output: Contribution to journalMeeting Abstract

TY - JOUR

T1 - Finding the brake

T2 - Neuro-Oncology

AU - Taylor, Sophie

AU - Knipp, Sabine

AU - Mavria, Georgia

AU - Ladbury, John

AU - Bruning-Richardson, Anke

PY - 2018/1

Y1 - 2018/1

N2 - Cell migration is one of the hallmarks of cancer. Cancer cells can adopt two main migratory strategies displaying either a mesenchymal or amoeboid phenotype. Targeting cell migration presents an opportunity in improving treatment of invasive and migratory tumours, however the cellular mechanisms that control the cell migration phenotypes in high-grade gliomas, such as Glioblastoma multiforme, are still not fully understood. In previous work we used 2D and 3D migration assays to screen a panel of inhibitors targeting the actin polymerisation pathway, a key regulator of cell migration in glioma. Amongst RhoA pathway inhibitors, which are key negative regulators of migration, we have identified one compound, CCG-1423, that uniquely failed to inhibit migration. CCG-1423 induces a reduced expression of the adhesion protein CCN1 by blocking nuclear import of MKL-1, a transcriptional co-activator. We suggest that CCG-1423 induces a mesenchymal transition, which permits continued migration. To confirm the effect of CCG-1423 on CCN1 levels we used ELISA to detect secreted CCN1 levels in culture supernatants in treated and non-treated U251 and U87 cells. CCG-1423 significantly reduced CCN1 levels in supernatant of both cell lines compared to untreated controls. Further biochemical analysis was performed to establish levels of MKL-1 in treated versus untreated cells and to elucidate its association with other proteins. Preliminary data indicate an increase in MKL-1 concentration in treated cells, consistent with a build-up of MKL-1 in the cytosol due to lack of nuclear import, as previously reported. Further detailed characterisation of the RhoA signalling pathway in glioma will provide an understanding of the role of MKL-1 in mesenchymal migration and the effect of CCG-1423 on mesenchymal switching. We will next characterise the interactions between CCG-1423 and MKL-1, and use pharmacological and/or genetic manipulation to refine combination treatments to target both mesenchymal and amoeboid cell migration.

AB - Cell migration is one of the hallmarks of cancer. Cancer cells can adopt two main migratory strategies displaying either a mesenchymal or amoeboid phenotype. Targeting cell migration presents an opportunity in improving treatment of invasive and migratory tumours, however the cellular mechanisms that control the cell migration phenotypes in high-grade gliomas, such as Glioblastoma multiforme, are still not fully understood. In previous work we used 2D and 3D migration assays to screen a panel of inhibitors targeting the actin polymerisation pathway, a key regulator of cell migration in glioma. Amongst RhoA pathway inhibitors, which are key negative regulators of migration, we have identified one compound, CCG-1423, that uniquely failed to inhibit migration. CCG-1423 induces a reduced expression of the adhesion protein CCN1 by blocking nuclear import of MKL-1, a transcriptional co-activator. We suggest that CCG-1423 induces a mesenchymal transition, which permits continued migration. To confirm the effect of CCG-1423 on CCN1 levels we used ELISA to detect secreted CCN1 levels in culture supernatants in treated and non-treated U251 and U87 cells. CCG-1423 significantly reduced CCN1 levels in supernatant of both cell lines compared to untreated controls. Further biochemical analysis was performed to establish levels of MKL-1 in treated versus untreated cells and to elucidate its association with other proteins. Preliminary data indicate an increase in MKL-1 concentration in treated cells, consistent with a build-up of MKL-1 in the cytosol due to lack of nuclear import, as previously reported. Further detailed characterisation of the RhoA signalling pathway in glioma will provide an understanding of the role of MKL-1 in mesenchymal migration and the effect of CCG-1423 on mesenchymal switching. We will next characterise the interactions between CCG-1423 and MKL-1, and use pharmacological and/or genetic manipulation to refine combination treatments to target both mesenchymal and amoeboid cell migration.

U2 - 10.1093/neuonc/nox238.095

DO - 10.1093/neuonc/nox238.095

M3 - Meeting Abstract

VL - 20

SP - 21

EP - 22

JO - Neuro-Oncology

JF - Neuro-Oncology

SN - 1522-8517

IS - S1

ER -