Differentially expressed proteins in glioblastoma multiforme identified with a nanobody-based anti-proteome approach and confirmed by OncoFinder as possible tumor-class predictive biomarker candidates.

Glioblastoma multiforme is the most frequent primary malignancy of the central nervous system. Despite remarkable progress towards an understanding of tumor biology, there is no efficient treatment and patient outcome remains poor. Here, we present a unique anti-proteomic approach for selection of nanobodies specific for overexpressed glioblastoma proteins. A phage-displayed nanobody library was enriched in protein extracts from NCH644 and NCH421K glioblastoma cell lines. Differential ELISA screenings revealed seven nanobodies that target the following antigens: the ACTB/NUCL complex, VIM, NAP1L1, TUFM, DPYSL2, CRMP1, and ALYREF. Western blots showed highest protein up-regulation for ALYREF, CRMP1, and VIM. Moreover, bioinformatic analysis with the OncoFinder software against the complete "Cancer Genome Atlas" brain tumor gene expression dataset suggests the involvement of different proteins in the WNT and ATM pathways, and in Aurora B, Sem3A, and E-cadherin signaling. We demonstrate the potential use of NAP1L1, NUCL, CRMP1, ACTB, and VIM for differentiation between glioblastoma and lower grade gliomas, with DPYSL2 as a promising "glioma versus reference" biomarker. A small scale validation study confirmed significant changes in mRNA expression levels of VIM, DPYSL2, ACTB and TRIM28. This work helps to fill the information gap in this field by defining novel differences in biochemical profiles between gliomas and reference samples. Thus, selected genes can be used to distinguish glioblastoma from lower grade gliomas, and from reference samples. These findings should be valuable for glioblastoma patients once they are validated on a larger sample size.

Effect of propofol and sevoflurane on the inflammatory response of patients undergoing craniotomy.

BACKGROUND: The purpose of this randomised, single-centre study was to prospectively investigate the impact of anaesthetic techniques for craniotomy on the release of cytokines IL-6, IL-8, IL-10, and to determine whether intravenous anaesthesia compared to inhalational anaesthesia attenuates the inflammatory response. METHODS: The study enroled 40 patients undergoing craniotomy, allocated into two equal groups to receive either sevoflurane (n = 20) or propofol (n = 20) in conjunction with remifentanil and rocuronium. The lungs were ventilated mechanically to maintain normocapnia. Remifentanil infusion was adjusted according to the degree of surgical manipulation and increased when mean arterial pressure and the heart rate increased by more than 30 % from baseline. The depth of anaesthesia was adjusted to maintain a bispectral index (BIS) of 40-60. Invasive haemodynamic monitoring was used. Serum levels of IL-6, IL-8 and IL-10 were measured before surgery and anaesthesia, during tumour removal, at the end of surgery, and at 24 and 48 h after surgery. Postoperative complications (pain, vomiting, changes in blood pressure, infection and pulmonary, cardiovascular and neurological events) were monitored during the first 15 days after surgery. RESULTS: Compared with patients anaesthetised with sevoflurane, patients who received propofol had higher levels of IL-10 (p = 0.0001) and lower IL-6/IL-10 concentration ratio during and at the end of surgery (p = 0.0001). Both groups showed only a minor response of IL- 8 during and at the end of the surgery (p = 0.57). CONCLUSIONS: Patients who received propofol had higher levels of IL-10 during surgery. Neither sevoflurane nor propofol had any significant impact on the occurrence of postoperative complications. Our findings should incite future studies to prove a potential medically important anti-inflammatory role of propofol in neuroanaesthesia. CLINICAL TRIAL REGISTRATION: Identified as NCT02229201 at www.clinicaltrials.gov.

Prognostic implication of preoperative behavior changes in patients with primary high-grade meningiomas.

High-grade meningiomas are rare extra-axial tumors, frequently causing brain invasion and prominent brain edema. Patients harboring high-grade meningiomas occasionally present with behavior changes. Data about frequency and prognostic importance of preoperative behavior changes in patients with high-grade meningiomas is missing. 86 patients with primary high-grade meningiomas were analyzed. Statistical analysis was performed to determine correlation of preoperative behavior changes with tumor location, preoperative brain edema, tumor cleavability, tumor grade, Ki67 proliferation index, and microscopic brain invasion. Survival analysis was performed. 30 (34.9%) patients presented with preoperative behavior changes. These changes were more frequent with male patients (P = 0.066) and patients older than 55 years (P = 0.018). They correlated with frontal location (P = 0.013), tumor size (P = 0.023), microscopic brain invasion (P = 0.015), and brain edema (P = 0.006). Preoperative behavior changes did not correlate with duration of symptoms, tumor cleavability, tumor malignancy grade, and Ki67 proliferation index. They were not significantly related to overall survival or recurrence-free survival of patients with primary high-grade meningiomas. Preoperative behavior changes are frequent in patients harboring primary high-grade meningiomas. They correlate with tumor size, microscopic brain invasion, and brain edema. Preoperative behavior changes do not predict prognosis in patients with primary high-grade meningiomas.

Caspase-3 and survivin expression in primary atypical and malignant meningiomas.

Objective. Information about possible prognostic factors of the survival of patients with atypical and malignant meningiomas (AMM) is sparse. The aim of our study was to evaluate prognostic significance of apoptotic marker caspase-3 and apoptotic inhibitor survivin in a series of primary AMM. Methods. 86 AMM (76 atypical and 10 malignant) were analyzed. Caspase-3 and survivin expression was evaluated immunohistochemically. The correlation between caspase-3, survivin, and other possible factors of meningioma recurrence was evaluated. Uni- and multivariate recurrence-free survival (RFS) and overall survival (OS) analyses were performed. Results. The intensity of caspase-3 expression correlated with the tumor grade (P = 0.004), the proliferation index (P = 0.019), and the mitotic count (P = 0.013). Survivin tended to be more expressed in female patients (P = 0.072). Survivin expression was stronger in malignant compared to atypical meningiomas, however, the difference was not statistically important (P = 0.491). Neither survivin nor caspase-3 expression significantly predicted OS or RFS in patients with AMM. Conclusions. Strong caspase-3 expression on AMM cells could reflect a cellular attempt at the homeostatic autoregulation of the tumor size. Survivin expression on AMM cells is similar to the survivin expression reported on benign meningiomas. Caspase-3 and survivin expression has no prognostic significance on the survival of patients with AMM.

New insights into meningioma: from genetics to trials.

PURPOSE OF REVIEW: In view of growing interest and need for nonsurgical therapies of meningiomas, we reviewed relevant articles published in the period from December 2010 to April 2012. RECENT FINDINGS: Large population-based, case-control studies have resulted in identification of new etiology factors, such as smoking or allergy, as well as new genes shown to play a role in meningioma susceptibility. Although aggressive surgery is still the treatment of choice for meningioma patients, even in the elderly population, postoperative radiotherapy remains debated and limited to Grade II residual cases as well as Grade III tumors. Although preclinical studies have pinpointed new candidate drugs to stop meningioma growth, such as trabectedin and histone deacetylase inhibitors, there are currently no effective therapies for meningiomas. Several clinical trials are under way, mostly on heavily pretreated patients, to determine the efficacy of some of the most promising candidate drugs, platelet-derived growth factor receptor-targeted therapies, and antiangiogenetic drugs being on the top list. Robust genetically engineered mouse models should be used as 'filters' to select agents for human clinical trials. SUMMARY: Surgical approach combined with radiotherapy in selected cases remains the most appropriate for meningioma patients. New therapies are under investigation and should modify treatment paradigms in the future.

New developments in surgery of malignant gliomas.

BACKGROUND: Malignant gliomas account for a high proportion of brain tumours. With new advances in neurooncology, the recurrence-free survival of patients with malignant gliomas has been substantially prolonged. It, however, remains dependent on the thoroughness of the surgical resection. The maximal tumour resection without additional postoperative deficit is the goal of surgery on patients with malignant gliomas. In order to minimize postoperative deficit, several pre- and intraoperative techniques have been developed. CONCLUSIONS: Several techniques used in malignant glioma surgery have been developed, including microsurgery, neuroendoscopy, stereotactic biopsy and brachytherapy. Imaging and functional techniques allowing for safer tumour resection have a special value. Imaging techniques allow for better preoperative visualization and choice of the approach, while functional techniques help us locate eloquent regions of the brain.

Mitotic count, brain invasion, and location are independent predictors of recurrence-free survival in primary atypical and malignant meningiomas: a study of 86 patients.

BACKGROUND: Since precise diagnostic criteria for atypical and malignant meningiomas (AMMs) were provided for the first time in the 2000 World Health Organization (WHO) criteria, there is only sparse information about possible prognostic factors in the group of AMMs. OBJECTIVE: To evaluate the prognostic significance of various histological and clinical parameters in AMMs, with an emphasis on location, mitotic count, brain invasion, and Ki67 labeling index. METHODS: We analyzed 86 primary AMMs, 76 of which were atypical and 10 of which were malignant, diagnosed according to the 2000 WHO classification. Multivariate Cox survival analyses were performed. RESULTS: High mitotic count, brain invasion, and the parasagittal-falcine location of the tumor were significantly associated with decreased recurrence-free survival in multivariate analysis. Brain invasion was present in 25 of 37 cases in which brain tissue was identified in the tumor specimens. When brain invasion was not included in the analysis because of the limited number of cases in which it could be assessed, high mitotic count, Ki67 index >4%, the presence of macronucleoli, and parasagittal-falcine location were significant predictors of shorter recurrence-free survival. CONCLUSION: AMMs, as defined by 2000 WHO, are biologically heterogeneous. Recurrence-free survival can be further stratified by location and histological parameters, especially mitotic count, brain invasion, and Ki67 labeling index. Not only brain invasion, but also the presence or absence of brain tissue in surgical specimens should be reported, because the absence of brain invasion, when brain tissue is identified, provides very important positive prognostic information.

Antigen expression on recurrent meningioma cells.

INTRODUCTION: Meningiomas are intracranial brain tumours that frequently recur. Recurrence rates up to 20% in 20 years for benign meningiomas, up to 80% for atypical meningiomas and up to 100% for malignant meningiomas, have been reported. The most important prognostic factors for meningioma recurrence are meningioma grade, meningioma invasiveness and radicality of neurosurgical resection. The aim of our study was to evaluate the differences in antigenic expression on the surface of meningioma cells between recurrent and non-recurrent meningiomas. METHODS: 19 recurrent meningiomas and 35 non-recurrent meningiomas were compared regarding the expression of MIB-1 antigen, progesterone receptors, cathepsin B and cathepsin L, using immunohistochemistry. RESULTS: MIB-1 antigen expression was higher in the recurrent meningioma group (p=0.001). No difference in progesterone receptor status between recurrent and non-recurrent meningiomas was confirmed. Immunohistochemical intensity scores for cathepsin B (p= 0.007) and cathepsin L (p<0.001) were both higher in the recurrent than in the non-recurrent meningioma group. CONCLUSIONS: [corrected] MIB-1 antigen expression is higher in recurrent compared to non-recurrent meningiomas. There is no difference in expression of progesterone receptors between recurrent and non-recurrent meningiomas. Cathepsins B and L are expressed more in recurrent meningiomas.