Laboratory Study
Prognostic significance of Tks5 expression in gliomas

https://doi.org/10.1016/j.jocn.2011.11.013Get rights and content

Abstract

A pathological hallmark of gliomas is their extensive invasion into the brain parenchyma regardless of tumour grade. Clinically this is a major factor in tumour recurrence as surgery and adjuvant therapies are unable to eradicate all the infiltrating malignant cells. Tyrosine kinase substrate with five SH3 domains (Tks5, also known as SH3PXD2A) and cortactin are required for the formation of invadopodia, actin-based protrusions of tumour cells with associated proteolytic activity implicated in tumour invasion. We investigated the prognostic significance of Tks5 and cortactin expression in 57 patients with various grades of glioma. Expression of Tks5 or cortactin occurred in all grades of tumours and expression of Tks5, but not cortactin, was associated with significantly reduced patient survival among glioma patients. This association was clearest in patients with low-grade astrocytomas and oligoastrocytomas. These results suggest a prognostic relevance for the Tks5 invadopodial protein in glial-derived brain tumours.

Introduction

Malignant gliomas caused approximately 2.3% of cancer-related deaths in the USA in 2010 with over 22,000 new patients expected annually.1 The most prevalent form of glioma and the tumour with the worst prognosis is the glioblastoma multiforme (GBM, World Health Organization [WHO] grade IV).2 Despite technological advances in neuroimaging, surgery and post-operative treatments, the prognosis for patients with GBM tumours remains poor.3, 4, 5 Individuals receiving the current standard treatment for high-grade gliomas, consisting of surgery, fractionated radiotherapy and concomitant chemotherapy with the DNA alkylating drug temozolomide, show a median survival of only 14.6 months with just 26.5% of individuals surviving for two years.6 The characteristics of all gliomas is their extensive infiltration, which thwarts efforts to completely remove or ablate malignant cells.5, 7, 8, 9 At an anatomical level, glioma invasion typically occurs via defined routes, with cells preferentially migrating along tracts of myelinated axons, the subependyma and the basement membranes of blood vessels.10

A property shared by several types of tumour cells with high invasive or metastatic potential is an ability to form invadopodia, dynamic actin-dependent protrusions which adhere to and proteolytically degrade extracellular matrix (ECM) substrates via the activities of numerous transmembrane and secreted extracellular proteases.11, 12, 13, 14 Functional (matrix-degrading) invadopodia have been observed both in glioma cell lines15 and primary tumour cells derived from ex vivo cultured GBM specimens,13 suggesting a possible role for invadopodia in glioma invasion.

The proteolytic function of invadopodia is reliant on the cooperative interactions of an underlying network of cell adhesion, signalling, adaptor and actin regulatory proteins.12, 13, 16 The Tks5 adaptor protein (also known as SH3PXD2A), which comprises a phox homology (PX), five src homology 3 (SH3) domains and Src phosphorylation sites17 is a marker of invadopodia.13, 18, 19, 20 Tks5 is critical for invadopodia formation and proteolysis-directed invasion of cells through ECM19 and facilitates growth of Src transformed fibroblasts in a mouse model of metastasis.21 Tks5 is recruited to invadopodia membranes by binding selectively to a subset of 3′-phosphatidylinositides via its PX domain.18, 22 Moreover, several proteins involved in actin and membrane regulation that are critical for invadopodia functional activity bind to Tks5 SH3 domains and/or Src phosphorylation sites (for example, tyrosine Y557) including N-WASP, dynamin and Nck adaptor proteins.22, 23, 24, 25 Recently, an essential role for a Src–Tks5 signalling pathway involving Tks5 tyrosine phosphorylation was uncovered in migration of neural crest-derived cells during zebrafish embryogenesis26 which supports the notion that developmental cell migrations may provide default mechanisms that drive invasion and metastasis.26 The transcription factor Twist1, which can stimulate the epithelial–mesenchymal transition, induces invadopodia formation and metastasis via a Src–Tks5 dependent pathway.27

The actin filament-binding protein cortactin, which additionally promotes actin polymerization and branching by activating the Arp2/3 actin nucleation complex, is also a central mediator of invadopodia formation.28, 29 Tyrosine phosphorylation of cortactin has been reported to mediate Nck1 binding and contributes to the initiation of invadopodia formation,20, 30 whereas its dephosphorylation stabilizes invadopodia by preventing cofilin-mediated actin filament severing.20 Cortactin expression has been linked to poor disease-specific survival in patients with breast carcinoma,31 melanoma32 and head and neck squamous cell carcinoma.33, 34 In the current retrospective study we evaluated the prognostic significance of the invadopodia-related proteins, cortactin and Tks5 in glioma patients.

Section snippets

Patients and specimen selection

Fifty seven surgically resected brain tumour specimens were retrieved from the Department of Surgery Brain Tumour tissue bank (The Royal Melbourne Hospital) at The University of Melbourne. Tumour specimens had been histologically classified at the time of storage by senior neuropathologists at the hospital according to WHO classification criteria. GBM (WHO grade IV, n = 20), anaplastic astrocytoma (AA, WHO grade III, n = 20) and astrocytoma and oligoastrocytoma (A/OA, WHO grade II, n = 17) specimens

Patient characteristics

Gliomas from 57 patients analysed in the present study comprised 20 GBM, 20 AA, and 17 A/OA, based on the WHO 2000 Classification of CNS tumours.2 The characteristics of the study participants are summarized in Table 1. The average age of patients at the time of surgery was 49 years (range 19–77 years), with 95% of the patients aged 70 years or younger. The ratio of male (31) to female (26) patients was approximately 1:1 and the mean KPS score of the patients, a measure of their general well-being

Discussion

The invasiveness of gliomas, irrespective of grade, is a major contributing factor to disease recurrence and patient mortality, albeit more rapid in patients with GBM compared with their lower grade counterparts.10 Although adjuvant treatment with photodynamic therapy (PDT), which targets infiltrating cells near the tumour bed, has shown improved patient outcome, there is a selection bias in these series.38, 39, 40

The WHO classification of gliomas, based on histopathology, is broadly effective

Acknowledgements

The authors gratefully acknowledge funding support from the National Health and Medical Research Council of Australia, Friends of the Royal Melbourne Hospital Neurosciences Foundation and the Cure-for-Life™ Foundation.

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