An Experimenter’s Guide to Glioblastoma Invasion Pathways

doi:10.1016/j.molmed.2018.07.003

Trends in Molecular Medicine

Volume 24, Issue 9, September 2018, Pages 763-780

https://doi.org/10.1016/j.molmed.2018.07.003 Get rights and content

Highlights

Glioblastoma (GBM) is a highly aggressive brain tumor that is characterized by an unparalleled invasion capacity.

GBM cells invade the brain via the brain parenchyma, the perivascular space, white matter tracts, and the leptomeningeal space.

GBM invasion signaling is dependent on the brain microenvironment, including extracellular matrix composition.

A pro-invasive GBM signaling network is emerging from literature revolving around Ephrin receptors, Rho GTPases, and casein kinase 2 that offers several druggable targets.

Glioblastoma is a highly aggressive brain tumor that is characterized by its unparalleled invasiveness. Invasive glioblastoma cells not only escape surgery and focal therapies but also are more resistant to current radio- and chemo-therapeutic approaches. Thus, any curative therapy for this deadly disease likely should include treatment strategies that interfere with glioblastoma invasiveness. Understanding glioblastoma invasion mechanisms is therefore critical. We discuss the strengths and weaknesses of various glioblastoma invasion models and conclude that robust experimental evidence has been obtained for a pro-invasive role of Ephrin receptors, Rho GTPases, and casein kinase 2 (CK2). Extensive interplay occurs between these proteins, suggesting the existence of a glioblastoma invasion signaling network that comprises several targets for therapy.

Section snippets

Invasion: An Ominous Hallmark of Glioblastoma

Glioblastoma (GBM; see Glossary) is the most common and fatal adult brain tumor and is classified as a grade IV glioma [1]. GBM is highly aggressive and characterized by an unparalleled invasion capacity. Current therapeutic strategies are insufficient to control the disease, as reflected by a dismal median survival of less than 15 months from diagnosis [2]. The standard of care consists of maximal surgical resection followed by radiotherapy and temozolomide chemotherapy. However, this strategy

Key Pathways in Glioblastoma Invasion

Glioblastoma invasion can be studied using a spectrum of models, with each having its own strengths and weaknesses: in vitro 2D (Box 2), in vitro 3D (Box 3), ex vivo (Box 4), and in vivo (Box 5). Although more complex models are expected to be more relevant physiologically, complexity usually comes at the cost of reduced throughput and less straightforward data interpretation. A good understanding of the advantages and limitations of each model (Table 1) and their suitability to study different

Druggability of GBM Invasion Pathways

Elucidation of GBM invasion pathways provides handles for designing therapeutic interventions. However, inhibiting GBM invasion in itself is unlikely to have considerable impact on survival. Extensive invasion has already occurred at the time of diagnosis, forming the seeds for later recurrence and just preventing invasion will not kill these cells. Thus, anti-invasion therapies need to be combined with other treatments. The benefit of such combinations may come from the creation of more

Concluding Remarks

In this feature review, we focused on signaling pathways for which the role in GBM invasion has been extensively validated using in vitro systems and ex vivo and in vivo assays. Several other pathways have been linked to GBM migration in vitro but require further research to demonstrate their in vivo relevance (Table S1 in the supplemental information online). Most of these pathways have traditionally been evaluated using 2D in vitro systems, although many are known to be involved in 3D

Glossary

Boyden chamber: in vitro invasion model that evaluates the movement of cells through a porous insert in a transwell system, either in absence or presence of extracellular matrices such as Matrigel.
Casein kinase 2: a constitutively active intracellular kinase that is most known for its role in regulating immunological signaling but is emerging as a player in GBM invasion signaling.
Ephrin receptors: receptor tyrosine kinases that convey intracellular signaling and play important roles in vascular

References (153)

R. Stupp
Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial
Lancet Oncol.
(2009)
L. Cheng
Elevated invasive potential of glioblastoma stem cells
Biochem. Biophys. Res. Commun.
(2011)
R.G. Verhaak
Integrated genomic analysis identifies clinically relevant subtypes of glioblastoma characterized by abnormalities in PDGFRA, IDH1, EGFR, and NF1
Cancer Cell
(2010)
H.S. Phillips
Molecular subclasses of high-grade glioma predict prognosis, delineate a pattern of disease progression, and resemble stages in neurogenesis
Cancer Cell
(2006)
A. Segerman
Clonal variation in drug and radiation response among glioma-initiating cells is linked to proneural-mesenchymal transition
Cell Rep.
(2016)
M.T. Milano
Patterns and timing of recurrence after temozolomide-based chemoradiation for glioblastoma
Int. J. Radiat. Oncol. Biol. Phys.
(2010)
A.C. Bellail
Microregional extracellular matrix heterogeneity in brain modulates glioma cell invasion
Int. J. Biochem. Cell Biol.
(2004)
J. Lee
Tumor stem cells derived from glioblastomas cultured in bFGF and EGF more closely mirror the phenotype and genotype of primary tumors than do serum-cultured cell lines
Cancer Cell
(2006)
B.W. Day
EphA3 maintains tumorigenicity and is a therapeutic target in glioblastoma multiforme
Cancer Cell
(2013)
L.-F. Wang
Increased expression of EphA7 correlates with adverse outcome in primary and recurrent glioblastoma multiforme patients
BMC Cancer
(2008)

E. Binda

The EphA2 receptor drives self-renewal and tumorigenicity in stem-like tumor-propagating cells from human glioblastomas

Cancer Cell

(2012)

M. Hatano

EphA2 as a glioma-associated antigen: a novel target for glioma vaccines

Neoplasia

(2005)

A.J. Ridley

Rho GTPase signalling in cell migration

Curr. Opin. Cell Biol.

(2015)

B. Salhia

The guanine nucleotide exchange factors trio, Ect2, and Vav3 mediate the invasive behavior of glioblastoma

Am. J. Pathol.

(2008)

N. Pencheva

Identification of a druggable pathway controlling glioblastoma invasiveness

Cell Rep.

(2017)

V. Amodeo

A PML/Slit axis controls physiological cell migration and cancer invasion in the CNS

Cell Rep.

(2017)

P.P. Scaglioni

A CK2-dependent mechanism for degradation of the PML tumor suppressor

Cell

(2006)

K.P.L. Bhat

Mesenchymal differentiation mediated by NF-kappaB promotes radiation resistance in glioblastoma

Cancer Cell

(2013)

N.K. Noren et al.

Eph receptor–ephrin bidirectional signals that target Ras and Rho proteins

Cell. Signal.

(2004)

M. Nakada

EphB2/R-Ras signaling regulates glioma cell adhesion, growth, and invasion

Am. J. Pathol.

(2005)

D. Herrmann

Protein kinase CK2 interacts at the neuromuscular synapse with Rapsyn, Rac1, 14-3-3γ, and Dok-7 proteins and phosphorylates the latter two

J. Biol. Chem.

(2015)

E.D. Apel

Phosphorylation of neuromodulin (GAP-43) by casein kinase II. Identification of phosphorylation sites and regulation by calmodulin

J. Biol. Chem.

(1991)

L.A. Dokas

Regulation ofin vitro phosphorylation of the casein kinase II sites in B-50 (GAP-43)

Brain Res.

(1998)

N. Matsuo

Characterization of STEF, a guanine nucleotide exchange factor for Rac1, required for neurite growth

J. Biol. Chem.

(2002)

S. Lakhe-Reddy

Beta8 integrin binds Rho GDP dissociation inhibitor-1 and activates Rac1 to inhibit mesangial cell myofibroblast differentiation

J. Biol. Chem.

(2006)

D.N. Louis

The 2016 World Health Organization Classification of Tumors of the Central Nervous System: a summary

Acta Neuropathol.

(2016)

M.D. Prados

Toward precision medicine in glioblastoma: the promise and the challenges

Neuro Oncol.

(2015)

K. Kallenberg

Glioma infiltration of the corpus callosum: early signs detected by DTI

J. Neurooncol.

(2013)

S. Bao

Glioma stem cells promote radioresistance by preferential activation of the DNA damage response

Nature

(2006)

M. Lun

The natural history of extracranial metastasis from glioblastoma multiforme

J. Neurooncol.

(2011)

M. Osswald

Brain tumour cells interconnect to a functional and resistant network

Nature

(2015)

V.A. Cuddapah

A neurocentric perspective on glioma invasion

Nat. Rev. Neurosci.

(2014)

P.G. Gritsenko

Interstitial guidance of cancer invasion

J. Pathol.

(2012)

S.S. Rao

Toward 3D biomimetic models to understand the behavior of glioblastoma multiforme cells

Tissue Eng. B Rev.

(2014)

A. Kania et al.

Mechanisms of ephrin–Eph signalling in development, physiology and disease

Nat. Rev. Mol. Cell Biol.

(2016)

J.P. Himanen

Repelling class discrimination: ephrin-A5 binds to and activates EphB2 receptor signaling

Nat. Neurosci.

(2004)

K.L. Todd

EphA4 regulates neuroblast and astrocyte organization in a neurogenic niche

J. Neurosci.

(2017)

J.-W. Jiao

Ephrins as negative regulators of adult neurogenesis in diverse regions of the central nervous system

Proc. Natl. Acad. Sci. U. S. A.

(2008)

M. Nakada

The phosphorylation of ephrin-B2 ligand promotes glioma cell migration and invasion

Int. J. Cancer

(2010)

L.F. Wang

Increased expression of EphA2 correlates with adverse outcome in primary and recurrent glioblastoma multiforme patients

Oncol. Rep.

(2008)

F. Liu

A genome-wide screen reveals functional gene clusters in the cancer genome and identifies EphA2 as a mitogen in glioblastoma

Cancer Res.

(2006)

H. Miao

EphA2 promotes infiltrative invasion of glioma stem cells in vivo through cross-talk with Akt and regulates stem cell properties

Oncogene

(2015)

J. Wykosky

EphA2 as a novel molecular marker and target in glioblastoma multiforme

Mol. Cancer Res.

(2005)

L. Teng

Ligand-dependent EphB1 signaling suppresses glioma invasion and correlates with patient survival

Neuro Oncol.

(2013)

S.D. Wang

EphB2 receptor controls proliferation/migration dichotomy of glioblastoma by interacting with focal adhesion kinase

Oncogene

(2012)

M. Nakada

The phosphorylation of EphB2 receptor regulates migration and invasion of human glioma cells

Cancer Res.

(2004)

Y. Tu

Expression of EphrinB2 and EphB4 in glioma tissues correlated to the progression of glioma and the prognosis of glioblastoma patients

Clin. Transl. Oncol.

(2012)

M. Nakada

Ephrin-B3 ligand promotes glioma invasion through activation of Rac1

Cancer Res.

(2006)

B. Krusche

EphrinB2 drives perivascular invasion and proliferation of glioblastoma stem-like cells

eLife

(2016)

J.J. Li

EphrinA5 acts as a tumor suppressor in glioma by negative regulation of epidermal growth factor receptor

Oncogene

(2009)

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Invadopodia associated Thrombospondin-1 contributes to a post-therapy pro-invasive response in glioblastoma cells
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A critical challenge in the treatment of glioblastoma (GBM) is its highly invasive nature which promotes cell migration throughout the brain and hinders surgical resection and effective drug delivery. GBM cells demonstrate augmented invasive capabilities following exposure to the current gold standard treatment of radiotherapy (RT) and concomitant and adjuvant temozolomide (TMZ), resulting in rapid disease recurrence. Elucidating the mechanisms employed by post-treatment invasive GBM cells is critical to the development of more effective therapies. In this study, we utilized a Nanostring® Cancer Progression gene expression panel to identify candidate genes that may be involved in enhanced GBM cell invasion after treatment with clinically relevant doses of RT/TMZ. Our findings identified thrombospondin-1 (THBS1) as a pro-invasive gene that is upregulated in these cells. Immunofluorescence staining revealed that THBS1 localised within functional matrix-degrading invadopodia that formed on the surface of GBM cells. Furthermore, overexpression of THBS1 resulted in enhanced GBM cell migration and secretion of MMP-2, which was reduced with silencing of THBS1. The preliminary data demonstrates that THBS1 is associated with invadopodia in GBM cells and is likely involved in the invadopodia-mediated invasive process in GBM cells exposed to RT/TMZ treatment. Therapeutic inhibition of THBS1-mediated invadopodia activity, which facilitates GBM cell invasion, should be further investigated as a treatment for GBM.
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Citation Excerpt :
This highly inducible transmembrane receptor is overexpressed in various solid tumors, including high-grade gliomas, especially in the mesenchymal GBM molecular subtype, which has a highly aggressive and invasive phenotype [163]. Fn14 activation in glioma cells triggers various downstream signaling pathways, including the NF-кB and Jak/STAT pathways, which are implicated as being critical drivers of tumor cell invasion in high-grade gliomas [164,165]. Furthermore, Fn14 expression is reported to be highly elevated in GBM cells near the tumor margins, in regions of high microvascular proliferation, and in recurrent tumors that have undergone standard-of-care treatments [166].
Glioblastoma (GBM) is the most common malignant adult brain cancer with no curative treatment strategy. A significant hurdle in GBM treatment is effective therapeutic delivery to the brain-invading tumor cells that remain following surgery within functioning brain regions. Developing therapies that can either directly target these brain-invading tumor cells or act on other cell types and molecular processes supporting tumor cell invasion and recurrence are essential steps in advancing new treatments in the clinic. This review highlights some of the drug delivery strategies and nanotherapeutic technologies that are designed to target brain-invading GBM cells or non-neoplastic, invasion-supporting cells residing within the GBM tumor microenvironment.

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Trends in Molecular Medicine

Feature ReviewAn Experimenter’s Guide to Glioblastoma Invasion Pathways

Highlights

Section snippets

Invasion: An Ominous Hallmark of Glioblastoma

Key Pathways in Glioblastoma Invasion

Druggability of GBM Invasion Pathways

Concluding Remarks

Glossary

Lancet Oncol.

Biochem. Biophys. Res. Commun.

Cancer Cell

Cancer Cell

Cell Rep.

Int. J. Radiat. Oncol. Biol. Phys.

Int. J. Biochem. Cell Biol.

Cancer Cell

Cancer Cell

BMC Cancer

Cancer Cell

Neoplasia

Curr. Opin. Cell Biol.

Am. J. Pathol.

Cell Rep.

Cell Rep.

Cell

Cancer Cell

Cell. Signal.

Am. J. Pathol.

J. Biol. Chem.

J. Biol. Chem.

Brain Res.

J. Biol. Chem.

J. Biol. Chem.

The 2016 World Health Organization Classification of Tumors of the Central Nervous System: a summary

Acta Neuropathol.

Toward precision medicine in glioblastoma: the promise and the challenges

Neuro Oncol.

Glioma infiltration of the corpus callosum: early signs detected by DTI

J. Neurooncol.

Glioma stem cells promote radioresistance by preferential activation of the DNA damage response

Nature

The natural history of extracranial metastasis from glioblastoma multiforme

J. Neurooncol.

Brain tumour cells interconnect to a functional and resistant network

Nature

A neurocentric perspective on glioma invasion

Nat. Rev. Neurosci.

Interstitial guidance of cancer invasion

J. Pathol.

Toward 3D biomimetic models to understand the behavior of glioblastoma multiforme cells

Tissue Eng. B Rev.

Mechanisms of ephrin–Eph signalling in development, physiology and disease

Nat. Rev. Mol. Cell Biol.

Repelling class discrimination: ephrin-A5 binds to and activates EphB2 receptor signaling

Nat. Neurosci.

EphA4 regulates neuroblast and astrocyte organization in a neurogenic niche

J. Neurosci.

Ephrins as negative regulators of adult neurogenesis in diverse regions of the central nervous system

Proc. Natl. Acad. Sci. U. S. A.

The phosphorylation of ephrin-B2 ligand promotes glioma cell migration and invasion

Int. J. Cancer

Increased expression of EphA2 correlates with adverse outcome in primary and recurrent glioblastoma multiforme patients

Oncol. Rep.

A genome-wide screen reveals functional gene clusters in the cancer genome and identifies EphA2 as a mitogen in glioblastoma

Cancer Res.

EphA2 promotes infiltrative invasion of glioma stem cells in vivo through cross-talk with Akt and regulates stem cell properties

Oncogene

EphA2 as a novel molecular marker and target in glioblastoma multiforme

Mol. Cancer Res.

Ligand-dependent EphB1 signaling suppresses glioma invasion and correlates with patient survival

Neuro Oncol.

EphB2 receptor controls proliferation/migration dichotomy of glioblastoma by interacting with focal adhesion kinase

Oncogene

The phosphorylation of EphB2 receptor regulates migration and invasion of human glioma cells

Cancer Res.

Expression of EphrinB2 and EphB4 in glioma tissues correlated to the progression of glioma and the prognosis of glioblastoma patients

Clin. Transl. Oncol.

Ephrin-B3 ligand promotes glioma invasion through activation of Rac1

Cancer Res.

Feature Review
An Experimenter’s Guide to Glioblastoma Invasion Pathways