High-grade Gliomas Research Articles

P16.10.A UNRAVELING THE CLINICAL IMPLICATIONS OF TRIM24::NTRK2 FUSION GENE IN PEDIATRIC HIGH-GRADE GLIOMA: A CASE STUDY

Abstract BACKGROUND High-grade gliomas are among the deadliest of all childhood cancers and the leading cause of cancer-related mortality in children. The standard treatment is resection followed by chemotherapy, most often temozolomide and radiation therapy. Unfortunately, there is no evidence that temozolomide influences survival in pediatric patients and only a limited number of new drugs have been identified for specific subgroups of tumors. The identification of fusion genes as possible drivers of the disease offers an opportunity for targeted therapies. We present a case of a six-year-old patient with a novel TRIM24::NTRK2 fusion positive epithelioid glioblastoma with subsequent relapses. METHODS We investigated the ramification of this fusion and evolution of the tumor through whole genome sequencing, RNA-sequencing and DNA methylation profiling, and studied the effect of the patient’s treatment using patient-derived cell cultures. RESULTS Our studies show that the TRIM24::NTRK2 fusion has oncogenic abilities but loses its importance as the tumor evolves. In addition, we demonstrate an increased drug resistance over time, further indicating the diminished driver function of the fusion. CONCLUSION The use of resequencing revealed parallel development of tumor subclones with different genomic alterations, highlighting the importance of genomic profiling from various parts of the tumor. FUNDING This work was supported by the Swedish Research Council, the Swedish Childhood Cancer Foundation, and the Swedish Cancer Society

Use of a Single-Fiber Optical Probe for the Detection of Tumor Fluorescence in High-Grade Glioma.

High-grade glioma (HGG) is the most common primary brain tumor in adults. The overall median survival is between 14 and 16 months. Fluorescence-guided surgery by detection of protoporphyrin IX (PpIX) fluorescence has been shown to improve the extent of resection, translating to improved progression-free survival. Microscope-based fluorescence detection techniques are associated with several pain points, some of which may be addressed by using contact-based spectroscopy. We aimed to investigate the accuracy of an optical fiber at detecting PpIX fluorescence by performing real-time spectroscopy in patients with HGG. Adult patients undergoing fluorescence-guided surgery for suspected HGG were recruited prospectively. Intraoperatively, samples from cortex, white matter, and tumor were taken. These samples were interrogated using standard white and blue light microscope techniques and the optical fiber. These specimens were then assessed by neuropathology to determine whether the tumor tissue was present within them. We collected 89 samples from 28 patients. There was an equal ratio of men to women, with a median age of 66.5 years. The accuracy of the probe for detecting PpIX fluorescence was 75.9%, compared with 56.6% for the operating microscope, with a significant improvement in sensitivity (χ2 = 11.84, P < .001). The optical fiber probe was more accurate than the operating microscope for detecting PpIX fluorescence. This technology has potential value in improving the accuracy of fluorescence-guided surgery and has potential practical benefits relating to surgical workflow.

Intracranial administration of anti-PD-1 and anti-CTLA-4 immune checkpoint-blocking monoclonal antibodies in patients with recurrent high-grade glioma.

Recurrent high-grade glioma (rHGG) lacks effective life-prolonging treatments and the efficacy of systemic PD-1 and CTLA-4 immune checkpoint inhibitors is limited. The multi-cohort Glitipni phase I trial investigates the safety and feasibility of intraoperative intracerebral (iCer) and postoperative intracavitary (iCav) nivolumab (NIVO) ± ipilimumab (IPI) treatment following maximal safe resection (MSR) in rHGG. Patients received 10mg IV NIVO within 24h before surgery, followed by MSR, iCer 5mg IPI and 10mg NIVO, and Ommaya catheter placement in the resection cavity. Biweekly postoperative iCav administrations of 1-5-10mg NIVO (cohort 4) or 10mg NIVO plus 1-5-10mg IPI (cohort 7) were combined with 10mg IV NIVO for 11 cycles. 42 rHGG patients underwent MSR with iCer NIVO + IPI. 16 pts were treated in cohort 4 (postoperative iCav NIVO at escalating doses) while 28 patients were treated in cohort 7 (intra and postoperative iCav NIVO and escalating doses of IPI). The most common TRAE was fatigue; no grade 5 AE occurred. Dose-limiting toxicity was grade 3 neutrophilic pleocytosis (4 pts) receiving iCav NIVO plus 5 or 10mg IPI. PFS and OS did not significantly differ between cohorts (median OS: 42 [95% CI 26-57] vs. 35 [29-40] weeks; 1-year OS rate: 37% vs. 29%). Baseline B7-H3 expression significantly correlated with worse survival. OS compared favorably to a historical pooled cohort (n = 469) of Belgian rHGG pts treated with anti-VEGF therapies (log-rank P = .015). Intraoperative iCer IPI + NIVO with postoperative iCav NIVO±IPI up to biweekly doses of 1mg IPI + 10mg NIVO is feasible and safe, showing encouraging OS in rHGG patients. ClinicalTrials.gov registration: NCT03233152.

Estimating Progression-Free Survival in Patients with Primary High-Grade Glioma Using Machine Learning

Estimating Progression-Free Survival in Patients with Primary High-Grade Glioma Using Machine Learning

M6A RNA METHYLATION AS A THERAPEUTIC TARGET IN HIGH GRADE GLIOMA

Abstract AIMS High grade glioma (HGG) is a devastating brain cancer that has poor prognostic outcomes. Very recently, the field of epitranscriptome has emerged as a novel druggable target for multiple diseases, including cancers. In the present study, we sought to investigate the functional importance of N6-methyladenosine (m6A) RNA methylation in the pathogenesis and drug resistance of paediatric and adult high grade glioma. METHOD To identify the expression of key m6A effectors, RT-qPCR and immunofluorescence were undertaken in patient derived HGG cell lines, primary low passage HGG cells and human tissues. The expression of these effectors was modified using siRNA-mediated knockdown and small molecules to identify their effects on key transcription factors. The consequences of modification on the ability of the cells to form neurospheres and invade was also assessed. Developing chemotherapy-resistant cell lines is undertaking to assess the effect of m6A RNA levels on the ability of HGG cells to adapt to environmental changes. Measuring the level of m6A-modified transcripts were achieved by developing LC-MS/MS and MeRIP (methylated RNA immunoprecipitation) qPCR techniques. RESULTS We found that key m6A effectors such as methyltransferase like 3 (METTL3), Wilms tumor 1-associating protein (WTAP) and fat mass and obesity-associated (FTO) are expressed at significantly higher level in paediatric and adult HGG models than in control at both gene and protein levels. Furthermore, these effectors regulate the expression of key transcription factors in self-renewal, cell cycle regulation and tumorigenesis. siRNA-mediated depletion of m6A effectors and inhibition of METTL3 using small molecules such as STM2457 and STM3006 reduce the capacity of cells to form neurospheres and invade as well as improving the sensitivity of HGG cells to chemotherapeutic agents. CONCLUSION Our findings suggest that m6A effectors are potential pro-oncogenic factors that, if druggable, will represent an advanced avenue in HGG therapy.

EVALUATING THE RELATIONSHIP BETWEEN RADIOTHERAPY CLINICAL TARGET VOLUME AND GAIT ACTIVITY IN HIGH GRADE GLIOMA

Abstract AIMS Walking speed or peak cadence serves as a potential indicator of overall health, particularly in older individuals, with documented predictive value for various health-related outcomes. The emergence of wearable technology for gait analysis has facilitated early disease detection and monitoring in conditions like Parkinson’s disease. This study aimed to explore the association between gait metrics and radiotherapy Clinical Target Volume (CTV) in participants with High-Grade Glioma (HGG) enrolled in the BrainWear study, which captures longitudinal physical activity data through wearable devices. METHOD 21 HGG participants provided accelerometer data before and after a course of radiotherapy. Using Spearman rank correlation analysis, we examined the relationship between CTV and gait features while considering the total radiotherapy dose and tumour location. Participant data was cut at the same time point of 14-days either side of the start or end of radiotherapy to ensure the data was aligned. RESULTS The analysis revealed a moderately negative correlation between CTV and peak cadence at the onset of radiotherapy. At the end of radiotherapy and in the two weeks following completion, both peak cadence in 1 minute and 30 minutes showed a stronger inverse correlation with CTV (Spearman’s ρ = -0.52 and ρ = -0.44 respectively, p-value &amp;lt;0.05). This suggests that higher CTV may be associated with decreased peak cadence, particularly at the time of expected radiotherapy toxicity. Subgroup analysis focusing on patients with CTV covering the motor cortex further illustrated a more significant inverse correlation between CTV size and peak cadence (Spearman’s ρ = -0.77, p-value &amp;lt;0.05). CONCLUSION This study highlights the importance of understanding the interplay between tumour treatment volume, location and physical function. Further exploration of CTV volume in relation to anatomical areas such as the motor cortex could provide deeper insights into this relationship and provide opportunities for personalised monitoring in HGG.

AN EX-VIVO SPECTRAL AND LIFETIME-DECAY ANALYSIS OF 5-ALA DERIVED PPIX flUORESCENCE: OBJECTIVE flUORESCENCE MAPPING IN GLIOMA SURGERY

Abstract AIMS 5ALA/Protoporphyrin IX (PpIX) fluorescence guided surgery (FGS) in high-grade glioma has been shown to increase resection and survival, despite this benefits are limited in low-grade glioma by low emission and subjective interpretation. Quantitative fluorescence could address these issues, however is challenging due to photobleaching and variations in tissue induced distortion and PpIX photochemical state that are poorly understood. Lifetime-decay measures intrinsic fluorophore properties and is not influenced by these factors, but is difficult to measure intra-operatively. We used paired optical and fluorescence spectroscopy with lifetime decay mapping to characterise PpIX emission within the glioma tissue microenvironment. METHOD Biopsies were collected from 20 patients undergoing glioma FGS, along with 4 controls from partial temporal lobectomies. Paired measurements of emission and lifetime were matched with optical property readings. Lifetime and emission ratios at 635 and 620nm were calculated (PpIX635/620), corresponding to the primary emission peak for the photochemical states of PpIX in tissue. Raw and optically corrected emission was correlated with lifetime decay. RESULTS 205 specimens were analysed by WHO grade (56 Grade4, 66 Grade3, 37 Grade2, 20 control). PpIX635/620 emission was 0.93 in control (CI 0.91-0.95), 1.03 in grade 2 tumours (CI 0.95-1.26), 1.56 in grade 3 (CI 1.22-1.91) and 6.99 (CI 5.78-8.20) in grade 4. PpIX635/620 lifetime correlated with emission (R2 =0.98, range 0.94-1.57). Correlation of emission with lifetime readings improved significantly following optical correction at 635nm (raw R2 =0.81, corrected R2 =0.90) and 620nm (raw R2 =0.44, corrected R2 =0.86). CONCLUSION PpIX620 and PpIX635 photochemical states are present in human glioma, with an increase in PpIX635/620 suggesting increased WHO grade. The ability to distinguish these forms holds the potential to provide real-time data for tumour differentiation and pathology. Accurate characterisation and correction of tissue induced distortion significantly improves the ability to detect, characterise and quantify PpIX intra-operatively.

DYSREGULATION OF M6A RNA METHYLATION, R-LOOPS AND THE DNA DAMAGE RESPONSE IN PAEDIATRIC HIGH-GRADE GLIOMAS

Abstract AIMS Paediatric high-grade gliomas (pHGG) are virtually incurable malignant brain tumours. The regulation of R- loops is necessary for ensuring genomic stability. The formation of R-loops at DNA damage sites promotes DNA repair via m6A methylation of the RNA strand. We aimed to investigate whether m6A methylation, R-loop homeostasis and the DNA damage response is dysregulated in pHGG and whether inhibitors, such as ATM inhibitors, could be potential therapeutics for pHGG. METHOD To assess the global levels of m6A, R-loops, yH2AX, total ATM and phosphorylated ATM in paediatric HGG cell lines compared to control human astrocytes, immunocytochemistry was initially performed. Immunocyto- chemistry was also performed to assess localisation between m6A and R-loops, R-loops and yH2AX, R-loops and total/phosphorylated ATM, m6A and total/phosphorylated ATM, and total/phosphorylated ATM and yH2AX in pHGG cell lines and human astrocytes. We are also exploring two ATM inhibitors, AZD1390 and AZD0156, and are currently investigating their effects on m6A methylation, R-loops and the DNA damage response in pHGG. As a molecular mechanism involving the tumour suppressor ARID1A was recently discovered that is crucial for maintaining genomic stability, we are also exploring the role of this protein in pHGG. We are also investigating the potential involvement of m6A readers in the pathogenesis of pHGG. RESULTS Immunocytochemistry revealed that dysregulation of global m6A, R-loop, yH2AX and ATM occurs in pHGG cell lines as higher levels of expression was generally observed compared to human astrocytes. Moreover, m6A, R-loops, yH2AX and ATM appear to colocalise as high Pearson’s correlation coefficient values were obtained. Experiments exploring ATM inhibitors, ARID1A and m6A readers in pHGG are currently in progress, however the results will be presented. CONCLUSION It appears that dysregulation of m6A methylation, R-loop homeostasis and the DNA damage response occurs in pHGG and that molecular mechanisms involving m6A methylation and R-loops could be impaired.

DTI fiber-tracking parameters adjacent to gliomas: the role of tract irregularity value in operative planning, resection, and outcome.

The goal of glioma surgery is maximal tumor resection associated with minimal post-operative morbidity. Diffusion tensor imaging-tractography/fiber tracking (DTI-FT) is a valuable white-matter (WM) visualization tool for diagnosis and surgical planning. Still, it assumes a descriptive role since the main DTI metrics and parameters showed several limitations in clinical use. New applications and quantitative measurements were recently applied to describe WM architecture that surround the tumor area. The brain adjacent tumor area (BAT) is defined as the region adjacent to the gross tumor volume, which contains signal abnormalities on T2-weighted or FLAIR sequences. The DTI-FT analysis of the BAT can be adopted as predictive values and a guide for safe tumor resection. This is an observational prospective study on an extensive series of glioma patients who performed magnetic resonance imaging (MRI) with pre-operative DTI-FT analyzed on the BAT by two different software. We examined DTI parameters of Fractional anisotropy (FA mean, min-max), Mean diffusivity (MD), and the shape-metric "tract irregularity" (TI) grade, comparing it with the surgical series' clinical, radiological, and outcome data. The population consisted of 118 patients, with a mean age of 60.6 years. 82 patients suffering from high-grade gliomas (69.5%), and 36 from low-grade gliomas (30.5%). A significant inverse relationship exists between the FA mean value and grading (p = 0.001). The relationship appears directly proportional regarding MD values (p = 0.003) and TI values (p = 0.005). FA mean and MD values are susceptible to significant variations with tumor and edema volume (p = 0.05). TI showed an independent relationship with grading regardless of tumor radiological features and dimensions, with a direct relationship with grading, ki67% (p = 0,05), PFS (p < 0.001), and EOR (p < 0.01). FA, MD, and TI are useful predictive measures of the clinical behavior of glioma, and TI could be helpful for tumor grading identification and surgical planning.

INTRAOPERATIVE MRI FOR BRAIN TUMOURS: A 15-YEAR RETROSPECTIVE REVIEW AT A SINGLE AUSTRALIAN INSTITUTION

Abstract AIMS Maximal safe resection is the objective of most neuro-oncological operations. The extent of resection can be maximized using imaging updated intraoperatively, including intraoperative MRI (iMRI) theatre, which may translate to improved overall survival. This study aimed to present our 15-year iMRI theatre experience and review its utility. METHOD A retrospective cohort analysis was conducted to include all intracranial tumour operations performed in the iMRI theatre (2007–2022). Cases were classified according to their histopathology: low- and high-grade gliomas (LGG and HGG respectively), meningiomas, pituitary adenomas, and metastases. Demographic, clinical, and histological data was extracted from SurgiNet and analyzed by using the WHO 2021 classification. The impact of intraoperative MRI on operative outcomes was reviewed. RESULTS A total of 478 neuro-oncology cases were identified, consisting of 194 pituitary adenomas (n= 69 cases via microscope), 162 HGG, 108 LGG (n= 7 cases for biopsy), 10 cerebral metastases, and 4 meningiomas. Intraoperative MRI theatre utilization averaged 41 cases per year throughout our 15-year experience. The median age was 47 years old (range 35-59 yo.) with a slight predominance in the female population (n= 298, 52%). We identified 85 (17.8%) re-do cases, which include: pituitary adenoma (n= 15, 17.6%), glioma (n= 68, 80%), and intracranial metastasis (n= 2, 2.4%). Of the 10 intracranial metastases, melanoma consisted of 5, lung carcinoma of 3, and renal cell- and colorectal-carcinoma represented 1 case, respectively. Intraoperative MRI confirmed complete resection in all meningioma cases. CONCLUSION In this series, iMRI has been utilised mostly for pituitary adenoma resection and gliomas. It provides near realtime intraoperative navigation and imaging to maximise the extent of resection, prevent injury to important structures and improve patient outcomes in tumour resections.

How 5-ALA enlightens neurosurgery – results of a single centre study on high-grade gliomas

How 5-ALA enlightens neurosurgery – results of a single centre study on high-grade gliomas

DIFFERENTIAL CELL SURFACE PROTEOMICS OF FLOTILLIN-1, HMGB1 AND AMINOPEPTIDASE N IN H3 MUTATED PAEDIATRIC HIGH-GRADE GLIOMAS

Abstract AIMS High-grade gliomas (HGG) are central nervous system tumour in children, where treatment options remain elusive, leaving children with a dismal prognosis. Histone H3.3 mutations, K27M and G34V/R have been identified as the two most common recurrent somatic mutations in paediatric HGG (pHGG). These mutations are present in 50% of pHGG compared to only 1% in adult gliomas. Our previous proteomic research on cell membrane protein through protein MS data analyses have identified different signatures of differentially expressed cell membrane proteins in H3-G34R/V mutated pHGG cells, in comparison to H3 wild type tumour cells in culture. Among these proteins we have identified flotillin-1, HMGB1 (High mobility group box 1) and aminopeptidase N. Flotillin-1 is associated with numerous malignancies and HMGB1 also functions as an oncogene and aminopeptidase N is a cell surface metalloprotease involved in tumour progression. Our aim is to study the expression and localisation of these cell surface proteins in H3 mutated and wild type tumour cells, to validate them as targets for developing novel therapeutics. METHOD Expression and localisation of flotillin-1, HMGB1 and aminopeptidase N in H3.3 mutated pHGG cell lines in comparison to H3 wildtype glioma and control cells was analysed, using different protein expression assays including immunofluorescence microscopy and western blotting. RESULTS Results showed statistically significant differential protein expression and localisation of these proteins. Flotillin-1 had increased cell membrane localization in H3 mutated cells. HMGB1 expression was increased, while aminopeptidase N expression was decreased in the H3 mutated cells in comparison to wildtype pHGG. CONCLUSION This new knowledge of the cell membrane proteomics of H3 mutated pHGG may help us utilise Flotillin- 1,HMGB1 and aminopeptidase N, as potential therapeutic targets for these unique H3 mutated children’s brain tumours.

FROM VEIN TO BRAIN: ENHANCING TRAffiCKING OF CAR T-CELLS IN DIFFUSE MIDLINE GLIOMA

Abstract AIMS H3K27M-altered diffuse midline glioma (DMG) is the deadliest childhood brain tumour, with treatment limited by anatomical location and chemoresistance; palliative radiotherapy remains the mainstay of treatment. Chimeric antigen receptor (CAR) T-cell therapy produces promising pre-clinical results in treating these tumours whilst leaving interspersed normal brain cells intact. Early clinical outcomes of GD2-CAR T-cell therapy for H3K27M-altered DMG demonstrate short-lived neurological and radiological improvement. Enhancing trafficking to and infiltration into the tumour core may enhance CAR T-cell efficacy. Here, advanced T-cell engineering modules are tested for their ability to improve CAR T-cell trafficking. METHOD A library of T-cell engineering modules was created each with a unique DNA barcode to allow identification of CAR T-cells expressing a given module. The library includes chemokine receptors (CCR) to improve tumour homing by chemotaxis towards tumour-secreted chemokines, adhesion molecules to increase extravasation across the blood-brain-barrier, and molecules to degrade upregulated tumour extracellular matrix (ECM) components to improve CAR T-cell motility through the tumour. Murine T-cells were co-transduced with a CAR and barcoded advanced T-cell engineering modules and administrated intravenously in syngeneic models of high-grade glioma (HGG)/DMG. Tumour, normal brain, bone marrow, spleen and liver tissue were collected at several time points post CAR T-cell administration to assess presence of CAR T-cells using genetic barcodes. RESULTS Co-expression of advanced T-cell engineering modules did not impair CAR T-cell function in vitro. CARs co-expressing selected CCRs were preferentially detected in tumour tissue in mice with GL261-EGFRvIII or H3.3K27Mp53LOFPDGFRAWT-engrafted tumours. Findings were further validated by flow cytometry detection of selected CCR-expressing CAR T-cells as compared to CAR T-cells without additional engineering components. Similar experiments for adhesion molecules and ECM modifiers are planned. CONCLUSION Results to date indicate that CAR T-cells co-expressing selected CCRs enhance their homing to HGG/DMG tumours, encouraging further in vivo research to assess impact on CAR T-cell efficacy.

IMPROVING THE EffiCIENCY OF THE NEURO-ONCOLOGY NEW PATIENT CLINIC AT THE ROYAL MARSDEN HOSPITAL

Abstract AIMS Patients seen for the first time in the neuro oncology outpatient unit are typically reviewed by a registrar, then a consultant and a clinical nurse specialist (CNS). This can lead to repetition in history taking and prolonged appointment times. We aimed to improve this by introducing a clerking proforma. METHOD Prospective audit of all new patients seen in clinic over a four month period. The total clinic appointment time was recorded for each patient. A proforma was designed to cover the main domains of presenting symptoms, especially focussing on tumour related symptoms, medications including dexamethasone and antiepileptic use, and ensured the appropriate information was given to the patient about the treatment proposed and driving restrictions. The proforma was introduced in to the clinic after 2 months and results analysed to see if a difference was made in terms of overall patient time in the clinic. Qualitative feedback gathered from the team members involved. RESULTS 10 patients were included prior to the use of the proforma and 9 patients afterwards. Patients presented with a mixture of high and low grade gliomas. Treatment options discussed included chemotherapy, radiotherapy, surveillance and best supportive care. The average time of patient stay in the department prior to the proforma was 107 minutes (range 65 to 165) and after the introduction of the proforma was 91 (range 40 to 110). The proforma was well received by the clinical team, it was felt useful to have a template to structure the new patient consultation, especially for more junior registrars, and the consultants were more confident that the appropriate information had been given. CONCLUSION Within the confines of a small sample size, the introduction of a proforma reduced the average new patient consultation time and standardised the clinical information collected.

ROR1 MRNA EXPRESSION IN GLIOMA - A NOVEL PROGNOSTIC BIOMARKER

Abstract AIMS Glioblastoma is the most common and aggressive primary brain tumour in adults. Despite maximal surgical resection followed by concomitant chemo-/radiotherapy, the three-year survival remains less than 5%. Receptor tyrosine kinase-like orphan receptor 1 (ROR1) overexpression is associated with poor prognosis in several cancers. ROR1 therapeutics are in Phase I/II clinical trials making it an exciting novel target with translational potential to explore for glioblastoma treatment. The aim of this study was to examine the association between ROR1 mRNA expression and overall survival (OS), by applying the WHO 2021 classification of transcriptomic glioma datasets. METHOD Clinical, histological, and molecular data were extracted from four independent glioma datasets (TCGA, CGGA, Rembrandt, and Gravendeel) via the GlioVis portal and analysed using the WHO 2021 classification. Only confirmed cases of astrocytoma, oligodendroglioma, or glioblastoma, and ROR1 mRNA expression data were included (2,216 cases). Kaplan-Meier survival curves were produced for OS with low/high grade glioma versus ROR1 mRNA expression. Log-rank (Mantel-Cox) multivariate regression was applied to analyse the effect of selected covariates on OS and ROR1 mRNA expression. RESULTS We observed a significant decrease in median OS in patients with high-ROR1 mRNA expression when compared to the low-ROR1 mRNA expression cohort (20 and 70 months, respectively). High-grade gliomas represented the highest ROR1 mRNA expression across the consortium, resulting in a poor median OS of 16 months. CONCLUSION This study represents the first report of an association between OS and the oncogenic role of ROR1 in glioma, a targetable cell surface marker for novel putative treatment in glioblastoma.

PROLONGED SURVIVAL IN IDH-WILDTYPE GLIOBLASTOMA: CASE REPORTS, LITERATURE REVIEW AND CURRENT PERSPECTIVES OF MOLECULAR BEHAVIOR

Abstract AIMS Glioblastoma remains one of the most aggressive primary brain tumours, with a median survival of around 12 months despite advances in treatment. However, rare cases of long-term survival challenge this prognosis. The evaluation of age, tumour location, surgical approach and strategies, adjuvant therapy, and molecular behavior provides valuable prognostic and predictive information in patients with glioblastoma. METHOD We present a case report of two patients diagnosed with wild-type glioblastoma who have survived over 10 years post-primary surgery. A review of epidemiological data, preoperative and postoperative radiological reports, surgical approach, and histology reports were studied. Additionally, we review the current literature to explore current nuances in High-Grade Glioma Wild Type prognosis and potential future directions. Three major medical database engines, PubMed (Medline), Scopus (ELSEVIER), and Cochrane were used to conduct a thorough literature search. RESULTS Both patients underwent maximal safe resection followed by the Stupp Protocol, comprising adjuvant chemotherapy with temozolomide and radiotherapy. Our case report highlights two exceptional cases of prolonged survival in Glioblastoma patients, both under 51 years-old at the moment of diagnosis, early diagnosis with isolated lobar lesions, no compromise of deep white matter, and achieving maximal safe resection and functional independence. Molecular analysis revealed specific genetic in common, including IDH Wild Type, p53 mutation, retained ATRX expression, TERT mutation, and MGMT promoter methylation, consistent with a favourable prognosis. Moreover, patients without recurrence often exhibit distinct molecular profiles, such as MGMT promoter methylation, suggesting a potential subtype associated with prolonged survival. CONCLUSION Our case reports and literature review highlight the possibility of prolonged survival in Glioblastoma patients. The age, tumor location at the time of diagnosis, comprehensive treatment approaches, may contribute to improved outcomes. Molecular profiling, particularly assessing genetic mutations and MGMT promoter methylation, holds promise for identifying patients with a higher likelihood of prolonged survival.

IDENTIFYING YB1 AND NPM1 AS PART OF THE MUTANT HISTONE 3 PROTEIN INTERACTOME IN H3 MUTATED PHGG

Abstract AIMS Paediatric high-grade gliomas (pHGG) are the leading cause of childhood cancer deaths, accounting for over 40% of deaths in the UK. pHGG tumours harbour histone H3 mutations, defining 50% of pHGG cases and the major mutations are H3.3K27M and H3.3G34R/V. Our previous proteomic research on H3 intercatomes, identified unique and common interacting binding partners of H3-G34R and H3-G34V mutant proteins relative to wild type H3 protein. Among these binding proteins we have focused on YB-1 and Nucleophosmin 1 (NPM1). YB-1, is multifunctional transcription factor and a well- characterised oncogenic protein. NPM1 is a molecular chaperone and frequently overexpressed in cancers. Our aim is to analyse the role of YB-1 and NPM1 in H3 mutated pHGG, in order to identify targeted therapeutic options for these difficult to treat tumours. METHOD H3 mutated glioma, H3 wild type pHGG and control cell lines were cultured and then subjected to comparative immunofluorescence microscopy, and western blot analyses to investigate YB-1 and NPM1 expression. RESULTS Results showed statistically significant altered protein expression and localisation of YB-1 and NPM1 in H3 mutated pHGG in comparison to wild type and control cells. YB-1 expression was increased in H3 mutated cells and NPM1 showed increased cytoplasmic localization in H3 mutated cells in comparison to H3 WT glioma and control cells. CONCLUSION Results of the study support that YB-1 and NPM1 are important interacting proteins in H3G34-mutated pHGGs and can be explored as potential therapeutic targets in H3 mutated pHGG.

Differentiation between high-grade gliomas and solitary brain metastases based on multidiffusion MRI model quantitative analysis.

Differentiating high-grade gliomas (HGGs) from solitary brain metastases (SBMs) using conventional magnetic resonance imaging (MRI) remains challenging due to their similar imaging features. This study aimed to evaluate the diagnostic performance of advanced diffusion models, such as neurite orientation dispersion and density imaging (NODDI) and mean apparent propagator magnetic resonance imaging (MAP-MRI), incomparison to traditional techniques like diffusion-weighted imaging (DWI), diffusion tensor imaging (DTI), and diffusion kurtosis imaging (DKI) for distinguishing HGGs from SBMs. In total, 17 patients with HGGs and 26 patients with SBMs were prospectively recruited based on the established inclusion and exclusion criteria. Structural MRI sequences and diffusion spectrum imaging (DSI) were utilized to assess quantitative parameter models, including NODDI, MAP-MRI, DWI, DTI, and DKI. Quantitative parameters were measured for both the tumor parenchymal area and the peritumoral edema area. The quantitative parameters of the two patient groups were compared using either the independent Student's t-test or the Mann-Whitney U test. The effectiveness of each model was evaluated using receiver operating characteristic (ROC) curves and calculating the area under the curve (AUC). Finally, the DeLong test was employed to compare the diagnostic performance of each model through pairwise comparisons of ROC curves. Isotropic volume fraction (Viso) based on NODDI; mean squared displacement (MSD) and the return to plane probabilities (RTPP) based on MAP-MRI; radial diffusivity (RDk) and mean diffusivity (MDk) based on DKI; and axial diffusivity (AD), radial diffusivity (RD), and mean diffusivity (MD) based on DTI of the peritumoral edema tumor were significantly different between HGGs and SBMs (p < 0.05). The optimal single discriminant parameters for each model are NODDI_Viso, MAP-MRI_MSD, DKI_MDk, and DTI_AD. Among these, the AUC of Viso (0.809) exceeds that of MSD (0.733), MDk (0.718), and AD (0.779). The combined model, which incorporates DTI_AD, DKI_RD, and NODDI_Viso, demonstrated superior diagnostic performance (0.897). Advanced diffusion MRI quantitative parameters derived from NODDI, such as Viso, have the potential to enhance the differentiation between HGGs and SBMs. The integrated utilization of these models is anticipated to enhance diagnostic accuracy and refine MRI protocols for brain tumor assessment.

Analysis of geographical distribution of high-grade glioma diagnoses in the Alessandria province: pilot study preliminary results

Background: one under-researched line of investigation is gliomas, which are a group of malignant brain tumours with highly poor prognoses. Despite efforts to identify environmental risk factors for gliomas, their etiology still remains unclear. It turns out that several future developments are needed in order to understand other potential risk factors. To this end, the primary objective of the present pilot study is to conduct an assessment of the spatial distribution of high-grade brain tumor cases in the province of Alessandria, where possible risk factors such as the presence of chemical industries and asbestiform fibers are found. Materials and Methods: the study sample consisted of all patients diagnosed with high-grade glioma between January 2018 and April 2023, residents in the province of Alessandria at the time of diagnosis and referred to the health facilities of the "Azienda Sanitaria Locale di Alessandria" and the "Azienda Universitario-Ospedaliera di Alessandria". Biographical and pathological history variables were collected, and finally, data were processed in aggregate and anonymized form. A total of 103 deceased patients were enrolled, stratified by age and type of diagnosis. Results: it was seen that the city with the most cases was Alessandria, with 27 diagnoses found during the period under review (26.2%), followed by cities of Acqui Terme, Casale Monferrato, and Novi Ligure with seven diagnoses (6.8%). Preliminary results presented in this paper were compared with data already published. Conclusions: analyses performed to arrive at an initial geographical distribution of high-grade glioma diagnoses are consistent relative to the population density of each municipality considered.

Ginsenosides as Promising Therapeutic Agents for Glioma: Mechanisms of Action and Future Perspectives

Abstract: Brain cancers, particularly gliomas, are a significant cause of mortality worldwide. Gliomas are primary tumors of the central nervous system (CNS) and are characterized by diverse clinical and biological features. Despite advancements in clinical approaches and surgical techniques, the treatment of high-grade gliomas still poses multiple challenges. This article focuses on a key active substance found in Panax ginseng called Ginsenosides. Ginsenosides belong to a specific class of triterpenoid saponins and have demonstrated various therapeutic effects, including neuroprotective, anticancer, anti-inflammatory, and neuroprotective functions. These compounds have shown potential in the treatment of gliomas and other cancers. Several pathways associated with ginsenosides, such as Rg3, Rh2, Rd, and Rb1, have been extensively studied, and these compounds have been proposed as potential targets in glioma treatment. The precise mechanisms of action of ginsenosides in gliomas are still being investigated, but their ability to modulate various signalling pathways and exert multiple therapeutic effects makes them promising candidates for further research and development. Clinical trials and additional studies are necessary to validate their therapeutic benefits and determine the optimal dosage, administration route, and potential combination with other treatment modalities. In summary, ginsenosides, the active compounds found in Panax ginseng, exhibit various therapeutic effects, including potential anti-cancer properties in gliomas. Their ability to modulate multiple pathways makes them promising targets for further research in the field of glioma treatment. However, more studies are required to establish their effectiveness and safety in clinical settings.