Laboratory StudyPrognostic significance of Tks5 expression in gliomas
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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2021, Neurobiology of DiseaseCitation Excerpt :Verhaaren et al. replicated the association with locus 17q25 (rs7214628, p= 2.7×10−19) and identified novel loci at10q24 (rs72848980, p= 1.6×10−9), 2p21 (rs11679640, p= 4.4×10−8), 1q22 (rs2984613, p= 2.0×10−8) and 2p16 (rs78857879, p= 1.5×10−8). The novel loci included genes that have been previously associated with AD (2p21, 10q24) (Tan et al., 2012; Harold et al., 2007), neuroinflammatory diseases (2p21) (Guillemin, 2012), intracerebral hemorrhage (1q22) (Woo et al., 2014), and glioma (10q24, 2p16) (Stylli et al., 2012; Hu et al., 2009). Recently, Lyall et al. (Lyall et al., 2020) identified an association between APOE ε4 and white matter hyperintensity volume (p= 0.001) by analyzing 8395 individuals with MRI data.
TKS5-positive invadopodia-like structures in human tumor surgical specimens
2019, Experimental and Molecular PathologyCitation Excerpt :The tools described here should allow a systematic analysis of different aspects of invadopodia-related biology in archived tumor samples. Increased levels of SH3PXD2A transcript (the gene encoding for TKS5) are associated with cancer progression and/or poor prognosis in a number of human malignancies including glioblastoma, schwannoma, and breast cancer (Blouw et al., 2015; Stylli et al., 2014; Stylli et al., 2012). TKS5 transcripts are overexpressed in pancreatic adenocarcinoma epithelium when compared to normal pancreas (Badea et al., 2008), but TKS5 protein levels in pancreatic adenocarcinoma have not been evaluated, and no information on transcript or protein levels is available for pancreatic neoplastic lesions with comparative better prognosis, such as papillary tumors (Dinarvand, 2017) or ampullary adenocarcinomas (Talamini et al., 1997).
Inhibition of Radiation and Temozolomide-Induced Invadopodia Activity in Glioma Cells Using FDA-Approved Drugs
2018, Translational OncologyCitation Excerpt :The increased mRNA expression of various invadopodia regulators in GBM tissue highlights the potential clinical significance of these structures in GBM patients. It also supports our previous evidence associating invadopodia with glioma cells [11,15]. Although the current therapeutic approach of radiotherapy and concomitant temozolomide postsurgery for GBM provides a modest improvement in survival, only 26.5% of individuals remain alive at 2 years due to tumor recurrence and invasion [40].
The emergent role of exosomes in glioma
2017, Journal of Clinical NeuroscienceCitation Excerpt :Importantly, the role of invadopodia in glioma invasion has been shown through the presence of functional (matrix-degrading) invadopodia in glioma cell lines [83] and primary tumour cells derived from ex vivo cultured GBM specimens [84]. Notably, Tks5 (also known as SH3PXD2A), which is an adaptor protein critical for invadopodia formation and proteolysis-directed invasion of tumour cells through the ECM [85–87], has been shown to possess prognostic potential in glioma patients with increased Tks5 expression resulting in significantly reduced survival among glioma patients [88]. Recently, it has also been demonstrated that the multiple invadopodia lifecycle steps, including their formation, stabilization and exocytosis of matrix degrading proteinases, may be mediated by the EVs known as exosomes [89].
Intercellular transfer of cancer cell invasiveness via endosome-mediated protease shedding
2024, Nature Communications
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These authors contributed equally to this work.