bluuebunny/crossref_metadata_2025 huggingface dataset:
KINOME-WIDE SYNTHETIC LETHAL SCREEN IDENTIﬁES PANK4 AS A MODULATOR OF TEMOZOLOMIDE RESISTANCE IN GLIOBLASTOMA
Authors: Vella Dr Viviana , Ditsiou Dr Angeliki , Chalari Dr Anna , Eravci Dr Murat , Wooller Dr Sarah K , Gagliano Prof Teresa , Bani Ms Cecilia , Kerschbamer Dr Emanuela , Karakostas Mr Christos , Xu Dr Bin , Zhang Dr Yongchang , Pearl Dr Frances M G , Lopez Dr Gianluca , Peng Dr Ling , Stebbing Prof Justin , Klinakis Prof Apostolos , Giamas Prof Georgios
Published: October 2024

AIMS

Temozolomide (TMZ) represents the cornerstone of glioblastoma (GBM) therapy. However, acquisition of resistance limits its therapeutic potential. The human kinome is an undisputable source of druggable targets, still, current knowledge remains conﬁned to a limited fraction of it, with a multitude of under-investigated proteins yet to be characterised. We aimed to uncover synthetic lethal partners of TMZ in GBM that could enhance the effect of TMZ, thus improving TMZ therapy response.

METHOD

A kinome-wide RNAi screen was performed in a TMZ-resistant GBM cell model. A panel of several TMZ-resistant and patient-derived GBM cell lines was implemented for in vitro validation of our ﬁndings through several phenotypic assays. In vivo TMZ-resistant GBM xenografts and immunohistochemical (IHC) analysis of IDH- wildtype GBM specimens were employed to assess the clinical relevance of our ﬁndings. Tandem Mass Tag (TMT)-based quantitative proteomic studies were undertaken to elucidate the underlying mechanisms.

RESULTS

Following a kinome-wide RNAi screen, pantothenate kinase 4 (PANK4) was uncovered as a modulator of TMZ resistance in GBM. Validation of PANK4 across various TMZ-resistant GBM cell models, patient-derived GBM cell lines, tissue samples, as well as in vivo studies, corroborated the potential translational signiﬁcance of these ﬁndings. Moreover, PANK4 expression is induced during TMZ treatment, and its expression is associated with a worse clinical outcome. A Tandem Mass Tag (TMT)-based quantitative proteomic approach revealed that PANK4 abrogation leads to a signiﬁcant downregulation of numerous proteins with central roles in cellular detoxiﬁcation and cellular response to oxidative stress. Mechanistically, as cells undergo genotoxic stress during TMZ exposure, PANK4 depletion represents a crucial event that can lead to accumulation of intracellular reactive oxygen species (ROS) and subsequent cell death.

CONCLUSION

Our study provides novel insights into chemoresistance in GBM and unveils a previously unreported role for PANK4 in mediating therapeutic resistance to TMZ in GBM.
OPTIC PATHWAY GLIOMAS AFTER TRANSITIONING INTO ADULTHOOD – AN INSTITUTIONAL EXPERIENCE
Authors: Loebel Dr Franziska , Aquilina Dr Kristian
Published: October 2024

AIMS

Optic pathway gliomas (OPG) are rare tumors affecting the optic nerve and the chiasm that occur predominantly during childhood and are generally managed by pediatric neurosurgeons/ pediatric oncologists. Prognosis is quite good for these benign tumors, and many patients survive into adulthood. While clinical care for these patients in childhood is well-standardized, the clinical course of patients with OPG after transitioning into adulthood is not well studied.We herein aimed to characterize patients with OPG diagnosed in childhood after transitioning into adult neurosurgical care in order in order to ellucidate the clinical course of OPG in adulthood and to identify potential problems with clinical care of these special patients in adult neurosurgery or neuro-oncology services.

METHOD

Clinical and radiographical data from adult patients (> 16 years of age) with OPG diagnosed in childhood and treated in our institution were collected from patient charts. Descriptive statistics and outcome analysis were performed using commercial statistical software.

RESULTS

Of 124 patients with OPG diagnosed in childhood, 35 patients transitioned into adulthood. The population included equal distribution of male (18, 51.4%) vs female (17, 48.6%) patients. Median age at diagnosis was 8.0 years (range 0.5 – 15.0) and median age at last follow up was 20.8 years (range 17.1 – 38.6). 12 patients were positive for NF (34.3%). 9 patients had undergone surgery for the OPG (25.7%), 11 had chemotherapy (31.4%) and 15 (42.8%) had received radiotherapy. 8 patients showed an endocrine deﬁciency (22.9%), 4 hypothalamic dysfunction (11.4%) and 8 cognitive/behavioral diﬃculties (22.9%). Visual acuity impairment was detected in 12 patients (34.3%). 9 patients experienced recurrence (25.7%).

CONCLUSION

Patients with OPG transition into adulthood frequently and experience a wide range of clinical problems. Close neurosurgical/neuro-oncology specialist follow-up is recommendable.
POSTOPERATIVE VISUAL ﬁELD DEﬁCITS ARE NOT ASSOCIATED WITH REDUCTION IN QUALITY OF LIFE IN GLIOBLASTOMA PATIENTS
Authors: Wan Mr Yizhou , Li Mr Yonghao , Sinha Mr Rohit , Jain Ms Swati , Mair Prof Richard , Morris Mr Robert , Joannides Mr Alexis , Santarius Mr Tom , Price Prof Stephen
Published: October 2024

AIMS

Preservation of functional integrity is a core goal of glioblastoma surgery. Visual ﬁeld deﬁcits can signiﬁcantly impact quality of life and patient function. Both tumour and surgical effects may affect visual function in these patients. The objective of this Study was to evaluate the effect of visual ﬁeld deﬁcits on HRQoL in the preoperative and early postoperative period (prior to chemoradiotherapy).

METHOD

48 patients with glioblastoma received preoperative, early postoperative (withon 1 week) and delayed postoperative testing (6 weeks) for visual function usinga combination of the OCS-Bridge neurocognitive screening tool and formal visual ﬁeld testing. Patients also completed the EORTC QLQ C30 with BN-20 and VFQ-25 visual function questionaires at the same timepoints. Statistical tests were performed using R Statistical Software (v4.1.2; R Core Team 2021).

RESULTS

The proportion of patients with visual ﬁeld deﬁcits at preoperative, early postoperative and late early postoperative timepoints were 20 (41.7%), 28 (58.3%) and 24 (50%) respectively. Most patients without a preoperative deﬁcit did not have an early 17/20 85% or late postoperative deﬁcit 19/24 79.1%. The proportion of patients with deﬁcits at early and late postoperative timepoints was signiﬁcantly higher in patients with preoperative deﬁcits compared to patients without preoperative deﬁcits, (Chi square p = 0.004) and (p = 0.008) respectively. There was no signiﬁcant differences between HRQoL measures between timepoints or between patients with deﬁcits at any timepoint compared to patients without deﬁcits.

CONCLUSION

Postoperative visual ﬁeld deﬁcits are related to preoperative tumour-effects rather than surgery. Patients can be counselled that while they do not recover following surgery, it is not associated with signiﬁcant reductions in HRQoL. Patients with preoperative visual ﬁeld deﬁcits may be considered for resection without signiﬁcant worry about the preservation of visual ﬁeld function since these deﬁcits do not recover postoperatively.
EXPLORING LOW-DENSITY LIPOPROTEIN RECEPTOR (LDLR) FAMILY IN GLIOBLASTOMA
Authors: Huyton Mr Jack , Smith Dr Christopher George Severin , Alder Dr Jane
Published: October 2024

AIMS

Cancer and neurodegenerative disease are unlikely to co-exist in the same patient. As glioblastoma is the most prevalent primary brain cancer, our research group is exploring how GBM is affected by genes implicated in Alzheimer’s disease. SorL1 has been established to be an important risk factor for Alzheimer’s disease and our group have shown it to be upregulated in patient derived GBM cells and it contributes to GBM cell migration and proliferation. SorL1 is part of a wider family of low-density lipoprotein receptor (LDLR) family and so my project was to explore the wider family as a potential target for investigation.

METHOD

The LDLR family were initially investigated using the bioinformatic data bases Xena browser and ivy Glioblastoma Atlas Project (ivyGAP). Xena browser reports RNA expression between healthy tissue samples (GTEX study) and from low grade glioma and glioblastoma and survival analysis while ivyGAP reports gene expression in the different anatomical regions of glioblastoma tumours.

RESULTS

Following bioinformatics and literature review: LRP1B, LRP5, LRP10 and Sortilin were chosen for further investigation. Western blot and immunocytochemistry were used to explore expression in cell lines SVGP12, U87MG and 1321N1. Sortilin and LRP10 were shown to be down regulated whilst LRP5 showed no signiﬁcant differences between cell lines.

CONCLUSION

The next step in this project is to explore expression of the LRP family in patient derived cells and normal human astrocytes. As the expression of the LDLR family was region speciﬁc across the tumour (ivyGAP data) future work in the project will explore expression in 3D spheroids and organotypic brain slice cultures, to create a model that better recapitulates the tumour microenvironment to better understand these proteins and their roles in GBM
DIFFERENTIAL CELL SURFACE PROTEOMICS OF FLOTILLIN-1, HMGB1 AND AMINOPEPTIDASE N IN H3 MUTATED PAEDIATRIC HIGH-GRADE GLIOMAS
Authors: Fairley Ms Hannah , Walugembe Ms Chloe , Sulman Mr Joshua , Haque Dr Farhana
Published: October 2024

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.
THE EPIGENETIC REGULATORY PROTEIN CBX2 AS A NOVEL THERAPEUTIC TARGET FOR GLIOBLASTOMA
Authors: Marsh Ms Danielle , Greenman Prof John , Wade Dr Mark
Published: October 2024

AIMS

Glioblastoma (GBM) is the deadliest and most common form of brain tumour. The average length of survival for GBM patients remains low, in part, due to a lack of targeted therapeutics. Identiﬁcation and validation of novel therapeutic targets is therefore crucial. Chomobox 2 (CBX2; a component of polycomb repressive complex 1 (PRC1)), represses gene expression and has a pro-oncogenic role in several aggressive cancer subtypes. CBX2 expression is elevated in GBM, however, very little is known about its role in this deadly disease. We aim to assess the phenotypic and gene expression effects of preventing CBX2-chromatin interaction in patient-derived GBM cell models, and to determine the functional mechanisms by which CBX2 promotes GBM progression; thereby determining its potential as a novel therapeutic target.

METHOD

Crucial to impactful target validation is the use of translationally relevant models, this study utilises a 3D spheroid model, using patient-derived cells, which more closely recapitulates the in vivo tumour microenvironment. GBM cell monolayers and spheroids were treated with SW2_152F, a selective CBX2 inhibitor. The phenotypic effects of CBX2 inhibition were assessed by cell count, CellTiter-Glo viability assays, ﬂuorescence microscopy, and ﬂow cytometry. Genes and biological pathways differentially regulated following CBX2 inhibition were assessed by RNA-sequencing and Gene Set Enrichment Analysis (GSEA).

RESULTS

Inhibition of CBX2 induced cell death and reduced cell viability in both 2D and 3D GBM cultures. RNA-seq and GSEA of SW2_152F-treated cells identiﬁed dysregulation of gene signatures involved in pro-oncogenic MYC and E2F signalling pathways, and dysregulation of genes associated with G2/M cell cycle progression.

CONCLUSION

Preventing CBX2-chromatin interaction induces GBM cell death. Further research will identify CBX2- chromatin binding sites to characterise the activity and direct regulatory role of CBX2 in GBM. Furthermore, we will assess the phenotypic and gene expression effects of CBX2 inhibition and depletion on ex vivo maintained GBM tissue biopsies.
QUALITY OF LIFE IN PATIENTS WITH SKULL BASE CHORDOMA AND CHONDROSARCOMA TREATED WITH PENCIL BEAM SCANNING PROTON THERAPY
Authors: Bryjova Dr Katarina , Mackeprang Dr Paul-Henry , Weber Prof Damien Charles
Published: October 2024

AIMS

The objective of this study is to assess the health-related quality of life (HRQoL) of skull base chor- doma/chondrosarcoma patients treated with proton therapy (PT).

METHOD

Seventy seven adult patients with skull base chordoma/chondrosarcoma completed the EORTC-QLQ-C30 and the BN20 Questionnaires prior to PT, Clinical outcome and therapy related acute and late toxicities have been recorded in course of the follow-up. The data was collected, categorized and analyzed, comparing the posttreatment scores to each patient’s baseline. In order to assess the impact of the clinical outcome and toxicities on HRQoL, the scores were correlated to collected clinical outcomes.

RESULTS

Median follow-up was 51 months. 6-year overall survival and local failure free survival rates observed were 85.1% and 80.3%. One G3 brain radiation necrosis and one G4 optic nerve disorder were documented during FU. HRQL deteriorated after the completion of proton therapy and returned to baseline values one year thereafter. After the second and third year, we observed further decrease in HRQL. This development was unrelated to tumor relapse. However, comparing the whole cohort over time, global QoL did not change substantially over the course of follow-up. GTR was correlated to better QoL at all time points. Disease progression was correlated with overall worse QoL and higher neurological symptoms already before PT, and higher symptom burden one year thereafter. Males had better QoL before and one year after PT than females.

CONCLUSION

PT achieves an excellent local and distant control in patients with skull base chordoma/chondrosarcoma. The HRQoL in these patients decreases immediately after the treatment completion, returns to the baseline from one year thereafter, and tends to deteriorate again with longer FU. The observed changes in global HRQoL were however not substantial in the whole cohort over the course of follow-up.
TARGETED THERAPY IN DIFFUSE INTRINSIC PONTINE GLIOMA: A COMPREHENSIVE REVIEW OF MONOCLONAL ANTIBODIES
Authors: Dib Ms Christie , Zaatar Mr Johnny , Sarouﬁne Ms Karmen , Elias Ms Linda , Alagha Mr Michel , Ghawi Ms Odette El , Khalil Ms Jana , Nakhle Mr Yves , Haidar Ms Elissa , Khoury Mr John-Victor El , Hawi Dr Jihad
Published: October 2024

AIMS

Diffuse intrinsic pontine glioma (DIPG) is a devastating pediatric brain tumor, marked by its poor prognosis and frequent recurrence despite conventional therapy. Our review examines the molecular landscape of DIPG, highlighting key factors driving tumor progression. We evaluate the therapeutic potential of various monoclonal antibodies (mAbs) targeting these factors, providing insights into their mechanisms and ongoing clinical trials. Our aim is to guide future investigations and immunotherapeutic strategies for DIPG.

METHOD

A narrative review was conducted using the PubMed database to identify relevant review articles, clinical trials, and observational studies published within the past 10 years, focusing on the use of mAbs in DIPG. The search terms “monoclonal antibodies”, “diffuse intrinsic pontine glioma”, and “targeted therapy” were employed. Additionally, a search was conducted on clinicaltrials.gov to gather information on ongoing and completed clinical trials.

RESULTS

Current human clinical trials on the checkpoint inhibitors anti-PD-1 (Nivolumab, Pembrolizumab, Balstilimab, Cemiplimab), anti-PD-L1 (Durvalumab), and anti-CTLA-4 (Ipilimumab, Zalifrelimab) have not demonstrated signiﬁcant improvement in overall patient survival. One major limitation has been the small sample size and limited expression of such markers in DIPG. Conversely, combination therapies involving anti-VEGF (Bevacizumab) and anti-EGFR (Nimotuzumab and Cetuximab) have shown promising results in small sample studies, but these results are not conclusive. Trials on anti-CD276 (Omburtamab) and CD40 agonist (APX005M) have shown no speciﬁc clinical or tumor outcomes in DIPG. Newer emerging agents like anti-TIM3 and anti-IL13Ra2 have shown very favorable results in in-vivo and in-vitro studies.

CONCLUSION

Current evidence is insuﬃcient to support the use of mAbs in DIPG. More large-size studies investigating the combinations of checkpoint inhibitors with different therapeutic agents are needed. Additionally, the promising results seen with anti-VEGF and anti-EGFR mAbs in their respective combination therapies warrant exploration in large-scale studies to draw more accurate conclusions. Furthermore, human trials should be conducted on anti-TIM3 and anti-IL13Ra2.
MULTI-OMIC INTEGRATION REVEAL INTRA-TUMOUR HETEROGENEITY AND IDENTIFY DISULﬁRAM AND COPPER AS A SYNERGISTIC THERAPY IN PAEDIATRIC EPENDYMOMA
Authors: Pandele Dr Alina , Whitby Dr Alison , Lankford Ms Sophie , Macarthur Mr Donald , Kamaly-Asl Mr Ian , Barrett Prof David , Grundy Prof Richard , Kim Dr Dong-Hyun , Rahman Dr Ruman
Published: October 2024

AIMS

Ependymoma (EPN) is the second most malignant paediatric brain tumour. The PF-A subgroup, associated with a hypoxic microenvironment, has a dismal survival rate of 50% with clinical trials failing to demonstrate a signiﬁcant survival advantage from chemotherapy to date. As the lack of genomic alterations makes it diﬃcult to uncover novel therapeutic targets, we present a multi-omic integration investigating whether spatially-distinct tumour microenvironments represent targetable metabolic niches in PF-A EPN.

METHOD

Surgical sampling of spatially-distinct regions was performed on eight PF-A patients. Metabolites and RNA were simultaneously extracted from the same cellular population and analysed using liquid chromatography-mass spectrometry (LC-MS) and RNAseq. Multi-omic integration was performed using Metscape3 and functional assays evaluating proliferation, invasion and siRNA-mediated knockdown were performed in 2D and 3D models of intra-tumour region patient-derived cell lines. Selected single and drug combination therapy was performed in normoxia and hypoxia, and synergy assessed using the Chou-Talalay method.

RESULTS

Multi-omic integration revealed 124 dysregulated metabolic pathways, encompassing 156 genes and 49 metabolites with the largest number of interactions occurring in the gluconeogenesis pathway. Each anatomical region also presented at least one unique gene-metabolite interaction demonstrating heterogeneity within and across PF-A EPN tumours. Four selected drug targets (Disulﬁram, Vandetanib, Mildronate and FBP1 inhibitor) based on metabolically relevant genes (ALDH3A1, FMO3, BBOX1, FBP1) showed impairment of metabolic viability in vitro, with limited chemosensitivity observed in control human cerebellar astrocytes. The addition of Cu2+ signiﬁcantly sensitized cells to Disulﬁram, particularly under hypoxic conditions. This synergistic combination signiﬁcantly impaired invasion in both normoxia and hypoxia.

CONCLUSION

This is the ﬁrst instance where multi-omic data integration and intra-tumour heterogeneity has been investigated for paediatric EPN revealing novel therapeutic targets in the context of gene-metabolite correlations. Drug tolerability will be further assessed in vivo using the previously characterised PF-A EPN orthotopic mouse xenograft MAF-928.
VASARI-AUTO: PERFORMANT, EQUITABLE, EﬃCIENT, ECONOMIC, AND SURVIVAL PREDICTIVE FEATURISATION OF GLIOMA MRI
Authors: Ruﬄe Dr James , Mohinta Ms Samia , Baruteau Dr Kelly Pegoretti , Rajiah Dr Rebekah , Lee Ms Faith , Brandner Prof Sebastian , Nachev Prof Parashkev , Hyare Dr Harpreet
Published: October 2024

AIMS

The VASARI (Visually AcceSAble Rembrandt Images) MRI feature set is a quantitative system designed to standardize glioma imaging descriptions. Though effective, deriving VASARI is time-consuming and therefore seldom used in clinical practice. This is however a problem that performant machine-learning software could plausibly automate.

METHOD

In 100 patients, two consultant neuroradiologists independently quantiﬁed VASARI features. In parallel, we developed VASARI-auto, an automated VASARI-labelling software applied to both open-source hand-segmented lesion masks and our openly available tumour segmentation model. We quantiﬁed: 1) agreement across neuroradiologists and VASARI-auto; 2) calibration of patient equity; 3) an economic workforce analysis; and 4) ﬁdelity in predicting patient overall survival. Neuroradiologists were blinded to VASARI-auto software development and evaluations, and software developers were blinded to neuroradiologist labelling.

RESULTS

Segmentation and VASARI-auto were equally performant regardless of age or sex (mean segmentation Dice coeﬃcient 0.947). A modest inter-rater variability between neuroradiologists (mean Cohen’s Kappa 0.49) was comparable between neuroradiologists and VASARI-auto (mean Cohen’s Kappa 0.41), with higher agreement across VASARI-auto methods (mean Cohen’s Kappa 0.94). The time taken for neuroradiologists to derive VASARI features was substantially higher than VASARI-auto (mean time per case 317 vs. 3 seconds, p<0.0001). A hospital workforce analysis of our centre forecast that three years of VASARI glioma featurisation would demand 771 consultant neuroradiologist workforce hours (£40,789) or 8.6 hours of computing time (£3.76 of power) with VASARI-auto. The best-performing survival model utilised VASARI-auto features (R2 0.25), opposed to those derived by consultant neuroradiologists (R2 0.21).

CONCLUSION

VASARI-auto is a highly eﬃcient automated labelling system for VASARI featurisation with equitable performance regardless of patient age or sex, a favourable economic proﬁle if used as a decision support tool, and with non-inferior ﬁdelity in downstream patient survival prediction. Future work should integrate such tools to enhance patient care.
INTEGRATING TRANSCRIPTOMICS WITH SPATIAL RADIOGRAPHIC ATLASES FOR GLIOMA ANALYSIS
Authors: Li Mr Yonghao , Wan Mr Yizhou , Price Prof Stephen
Published: October 2024

AIMS

The localization of gliomas in the brain is a critical prognostic factor, reﬂecting the genetic makeup of their originating cells. Variations in transcriptomic proﬁles across different regions and subtypes may further shed light on the mechanisms driving tumor development. Our research delves into the complex interplay between glioma locations and their transcriptomic signatures, seeking to dissect the spatial and genetic intricacies contributing to glioma diversity.

METHOD

In this study, we conducted a comprehensive analysis of preoperative anatomical MRI and transcriptomic data from 242 glioma patients to investigate the relationship between spatial distribution and tumor transcriptomic proﬁles. The MRI data were standardized by registration to the MNI-152 brain space. Tumors were segmented, and the brain was partitioned into 84 discrete subregions for detailed examination. Transcriptomic data underwent preprocessing, with principal component analysis employed to reduce dimensionality. Statistical analyses were used to assess the relationship between glioma description across different subtypes and PCA results. Additionally, we identify differentially expressed genes and common genes across regions for various glioma subtypes. Comparative analyses with the Allen Brain Atlas (divided into 84 subregions with expression data) were also performed to discern transcriptional differences between healthy individuals and glioma patients, further enhancing our understanding of glioma biology at the molecular level.

RESULTS

We generated 21 frequency maps detailing regional glioma occurrences, categorized by biomarkers, subtypes, and grades. Notably, some of these maps showed signiﬁcant correlations with expression principal component analyses (PCAs). Our ﬁndings include a set of genes consistently expressed across subregions, alongside region speciﬁc differentially expressed genes. Additionally, we identiﬁed distinct gene expression patterns when comparing glioma patients to healthy controls in the Allen Brain Atlas, highlighting potential molecular underpinnings of glioma heterogeneity.

CONCLUSION

Our analysis revealed notable correlations between glioma subtype locations and transcriptomic proﬁles, underscoring the role of transcriptomic signatures in explaining regional variations in glioma incidence.
SURGICAL OUTCOMES OF TUBULAR RETRACTOR SYSTEMS IN THE MANAGEMENT OF BRAIN TUMOURS: A SYSTEMATIC REVIEW AND META-ANALYSIS
Authors: Fard Mr Amir Rafati , Mowforth Mr Oliver , Hibberd Dr Owen , Akinduro Mr Isaac , Smith Mr Kieran , Patel Dr Reece , Bhatti Mrs Zainab , Karmarkar Mrs Sejal , Hill Mr Ciaran
Published: October 2024

AIMS

Retraction is a critical step in surgical removal of brain tumours, enabling visualisation and resection. Conventional retraction applies focal pressure to overlying tissue, potentially resulting in retraction-induced injury. Tubular retractors (minimally invasive parafascicular surgery) circumferentially distribute pressure to theoretically minimise injury. Whilst previous reviews have provided insight into their potential, the literature has since signiﬁcantly expanded, necessitating an update. The objective was to synthesise all current evidence regarding surgical outcomes of tubular retractors in resection of brain tumours.

METHOD

Studies reporting on use of tubular retractors to resect brain tumours in adult patients were eligible. PRISMA guidelines were followed. MEDLINE, Embase and the Cochrane Library were searched to 18th October 2023. Duplicate title/abstract screening, data extraction and risk of bias assessments were conducted.

RESULTS

Of the 2453 studies screened, 49 were included in the ﬁnal synthesis, involving 690 patients. Tubular retractor systems assessed were: BrainPath (n=199), VBAS (n=198), METRx (n=63) and other (n=230). Pooled estimate of gross total resection rate was 75% (95% CI 0.65-0.84; I2=83%), whilst pooled estimate of complication rate was 17% (95% CI 0.11-0.24; I2=75%). Sub-group analysis of gross total resection rate was conducted for: glioma (150 patients, pooled prevalence 47%, 95% CI 0.36-0.57; I2=29%), colloid cyst (121 patients, pooled prevalence 100%, CI 0.99-1.00; I2=0%), and metastasis (114 patients, pooled prevalence 82%, 95% CI 0.67-0.94; I2=64%). There was no signiﬁcant difference between tubular retractor systems with regards to gross total resection or complication rate (p>0.05).

CONCLUSION

There is mounting evidence for the utility of tubular retractors in resection of brain tumours. However, this evidence remains largely in the form of low-quality case-series, providing proof-of-concept, but limited in its ability to draw more robust comparisons to conventional retraction. Future studies with greater sample sizes and inclusion of control groups are now needed to consolidate the position of tubular retractors in neuro-oncology.
DESIGN AND SET UP OF A RANDOMISED CONTROLLED TRIAL OF MEDICINAL CANNABINOIDS IN RECURRENT GLIOBLASTOMA - THE ARISTOCRAT TRIAL
Authors: Short Prof Susan , Mant Dr Rhys , Savage Dr Josh , Patel Dr Amit , Billingham Prof Lucinda , Meade Dr Sarah , Buckle Mr Peter , Collinson Dr Fiona , Boele Dr Florien
Published: October 2024

AIMS

ARISTOCRAT is a unique, randomised, double-blind, placebo-controlled phase II trial of cannabinoids (nabixo- mols) alongside temozolomide in patients with MGMT-methylated recurrent GBM and is open to recruitment in 12 centres across the UK. It was rapidly funded through a crowdfunding campaign organised by The Brain Tumour Charity during the COVID-19 pandemic.

METHOD

This pragmatic phase II randomised trial was designed and set up by a multi-disciplinary team of co-applicants and trial management group (TMG) members comprising of clinicians, statisticians, trialists, Patient & Public Involvement (PPI), pharmacists and quality of life (QoL) researchers. The protocol randomises on a 2:1 ratio between nabiximols or matched placebo with standard dose temozolomide in patients at ﬁrst recurrence of IDH wild-type GBM. Unique aspects include self-titration of nabiximols based on individual tolerance, and fully blinded randomisation with matched placebo. The primary outcome measure is overall survival time with key secondary outcomes including survival at 6, 12 and 24 months, progression-free survival time, health-related QoL and adverse events. Intermediate assessment of feasibility to conﬁrm safety, compliance and achievability of recruitment will be undertaken after 40 patients are randomised. PPI input has been critical in the trial and patient-facing information design.

RESULTS

The trial has promoted a high level of patient and public interest, including press coverage. The trial aims to recruit a total of 234 patients over an 18 month period.

CONCLUSION

ARISTOCRAT will provide unique, robust clinical data on the role of nabiximols (combined with temozolomide) in improving overall survival and the impact on QoL for patients with recurrent GBM and is the ﬁrst of its kind in cancer patients.
IMPACT OF DNA METHYLATION PROﬁLING IN REﬁNING MENINGIOMA RECURRENCE RISK STRATIﬁCATION AND CLINICAL FOLLOW UP
Authors: Bakhsh Mr Ali , Brandner Prof Sebastian , Scott Prof Ian , Jenkinson Prof Michael , Rathi Dr Nitika
Published: October 2024

AIMS

To assess clinical impact of the additive information gained from Integrated Meningioma Risk Score on the risk of meningioma recurrence.

METHOD

DNA methylation (Illumina EPIC Array) analysis was performed on all recurrent meningioma tumours, primary tumours CNS5 WHO grade 2, 3 and grade 1 with unusual morphological features from Dec 2021 to Jan 2024. DNA methylation analysis was performed, and data ﬁles were processed on the DKFZ Heidelberg Classiﬁer v12.5/6 (molecularneuropathology.org). The output from the classiﬁer including methylation class combined with WHO Grade and chromosomal loss (1p, 6q,14q) was used to produce an integrated meningioma risk score for recurrence.

RESULTS

70 meningioma cases were proﬁled. This included 68 cranial meningiomas, ((WHO grade 1, n=19; recurrences=7), (grade 2, n=47; recurrences =9), (grade 3 n=2; recurrences=1)) and 2 spinal meningiomas (grade 1 n=1, grade2 n=1). 3 tumours were excluded due to poor calibration scores. Of the 67 cases the diagnosis was revised in 1 case following DNA methylation analysis due to presence of CDKN2A/B loss. Molecular-morphologic integrated risk score was calculated for all 67 cases. In 10% of overall patients the predicted risk of meningioma recurrence was increased to either intermediate or high, whilst 32% had the risk lowered mainly from intermediate to low. Interestingly, 1 patient who underwent a gross total resection of a WHO Grade 2 meningioma, experienced recurrence with the integrated meningioma risk score indicating low risk of recurrence, which is contrary to the clinical aggressiveness of the tumour.

CONCLUSION

Meningioma integrated risk stratiﬁcation by DNA methylation proﬁling has shown concordance of integrated risk score with WHO Grade in 58% of cases. In 42% of cases the predicted risk was altered when compared to the WHO Grade alone. These results need to be further validated with long term data on patient outcomes.
PROTAC’ING CENTROSOME AMPLIﬁCATION PROTEINS: TARGETED PROTEIN DEGRADATION AS A NOVEL THERAPEUTIC STRATEGY IN GLIOBLASTOMA
Authors: Thomaz Dr Amanda , Fielding Dr Andrew
Published: October 2024

AIMS

Glioblastomas (GBMs) are the most common and aggressive brain tumours, notoriously diﬃcult to treat with universal treatment resistance. A key feature of cancer cells is centrosome ampliﬁcation (CA), leading to chromosomal instability and worse patient outcomes, including metastasis development and treatment resistance. In this work, I identiﬁed CA proteins overexpressed in GBM and aimed to investigate how the targeted degradation of proteins associated with centrosome ampliﬁcation can inﬂuence the growth and survival of GBM cells.

METHOD

Based on well-established literature data concerning CA-related genes, I evaluated the expression level of these targets in different types of samples from the Tumour Cancer Genome Atlas dataset of GBM patient samples. Following the identiﬁcation of CA targets differentially overexpressed, to test the hypothesis that CA proteins could be involved in the survival of GBM cells and be potential therapeutic targets, I used a novel drug modality, named Proteolysis Targeting Chimeras (PROTACs) to degrade two CA proteins. PROTACs can quickly destroy speciﬁc proteins, potentially overcoming resistance to treatment. Target degradation was conﬁrmed via immunoblotting and cell viability and cell cycle analysis were performed to evaluate the impact on GBM cells.

RESULTS

Both primary and recurrent GBM samples display high levels of CA gene signature when compared to noncancerous samples, and some CA genes are particularly upregulated in recurrent samples. The PROTACs were effective in degrading both CA targets in GBM cells, leading to a signiﬁcant decrease in cell viability and G2 cell cycle arrest.

CONCLUSION

This study presents initial evidence that CA protein degradation could be a promising avenue for GBM research and therapy. Further research is needed to fully assess the translational potential of PROTACs in GBM treatment.
THE EﬃCACY OF GAMMA KNIFE RADIOSURGERY FOR CYSTIC METASTASIS COMPARED WITH NON-CYSTIC METASTASIS
Authors: Sheriff Dr Adam , Takami Prof Hirokazu , Tanaka Prof Shota , Nomura Prof Masashi , Takayanagi Prof Shunsaku , Hasegawa Prof Hirotaka , Umekawa Prof Motoyuki , Saito Prof Nobuhito
Published: October 2024

AIMS

Between 20-40% of patients with cancer develop brain metastases (BMets), with metastasis representing the most common type of brain tumour. Cystic metastasis describes a rarer type of BMet characterised by oedematous collections. Stereotactic radiosurgery (SRS) is an effective and minimally invasive method for treating non-cystic BMets but controversy exists regarding its use in cystic BMets. The traditional view that cystic lesions are less responsive to SRS however is not fully evidenced. This investigation aims to elucidate the effectiveness of Gamma Knife (GK) radiosurgery in achieving local control of cystic BMets and enhancing survival versus GK radiosurgery for non-cystic BMets.

METHOD

Patients who had undergone fractionated GK for either a non-cystic or cystic BMet between 2018 and 2021 were identiﬁed. BMet volume on pre-GK MRI was calculated. Information was additionally collected on BMet location, maximum cyst diameter, if present, the patient’s primary tumour and its control and their performance status at the time of GK. The follow-up of patients is ongoing with survival being calculated using the Kaplan- Meier method and local progression through volumetric analysis of repeat MRI scans.

RESULTS

57 patients with 79 cystic BMets were identiﬁed and 67 patients with non-cystic BMets. The most common primary tumour in both the non-cystic and the cystic BMets group was lung, comprising 30% and 23% respectively. The second most common tumour amongst non-cystic BMets was renal (12%) however in the cystic cohort it was breast (9%). The median tumour volume before treatment was 6.2cm3 in the non-cystic group and 12.5cm3 amongst cystic BMets. Median cyst size was 17.4 mm. Karnofsky Performance Status (KFS) was similar across both groups.

CONCLUSION

By comparing outcomes following GK for cystic and non-cystic metastasis, with the same primary tumours, this study can uncover the eﬃcacy of fractionated GK for cystic lesions and aid the selection of treatments for these patients.
THE IMPACT OF THE FRONTAL ASLANT TRACT ON POST-OPERATIVE OUTCOMES IN GLIOMA SURGERY
Authors: Rajwani Mr Kapil , Lavrador Mr Jose , Khan Mr Azharul , Banna Dr Qusai Al , Mirza Mr Asfand , Aliaga-Arias Mr Jahard , Elhag Mr Ali , Marchi Mr Francesco , Shapey Mr Jonathan , Bhangoo Mr Ranjeev , Ashkan Prof Keyoumars , Gullan Mr Richard , Vergani Mr Francesco
Published: October 2024

AIMS

The frontal aslant tract (FAT) has a role in speech and language functions and visual-motor activities. It is also not clear whether location of FAT relative to gliomas affects post-operative motor and language outcomes. Our hypothesis is FAT protects the cortico-spinal tract and other subcortical white matter tracts involved in speech.

METHOD

We conducted a retrospective cohort study. We used electronic patient records to search for all patients who had frontal tumours resected between Jan 2018 and August 2022. We included all adult patients (>18 years) with gliomas located in the SMA region and dorsal premotor cortex who had pre-op diffusion tensor imaging and underwent debulking / resection surgery. The patients were divided into 3 groups based on the location of the tumour relative to FAT. In group 1 patients had tumours located anterior to FAT, in group 2 patients had tumours inﬁltrating FAT and in group 3 patients had tumours located posterior to FAT. Our outcome measure was new post-operative language or motor deﬁcit.

RESULTS

Our search yielded 197 patients, of these 43 fulﬁlled the inclusion criteria. Sixteen patients were included in group 1, 9 patients included in group 2 and 18 included in group 3. Pre-operative motor deﬁcit was signiﬁcantly more common in patients with tumours posterior to FAT (p=0.046). Post-operative motor deﬁcit at hospital discharge was also more common on this group of patients (p=0.048). At 6 months follow up there was no signiﬁcant difference post-operative motor and language deﬁcits between the groups but there was a trend towards worse motor outcomes in patients with tumour located posterior to FAT.

CONCLUSION

There is a suggestion motor outcomes are worse in tumours located posterior to FAT. This can be considered when counselling patients for surgery with tumours related to FAT.
WHOLE BRAIN RADIOTHERAPY FOR INTRACRANIAL METASTATIC DISEASE IN NON-SMALL CELL LUNG CANCER: THE WESSEX EXPERIENCE
Authors: Starkey Dr Eleanor , Bhatnagar Dr Adityanarayan , Bates Dr Andrew , Fenton Dr Paul , Wood Dr Victoria , Begum Ms Nelima , Main Dr Sean
Published: October 2024

AIMS

The QUARTZ trial (2016) reported that corticosteroid therapy was non-inferior to whole brain radiotherapy(WBRT) for non-small cell lung cancer (NSCLC) with regards overall survival(OS). This is a single centre experience of WBRT for NSCLC in the post-QUARTZ era and in the context of the changes in clinical practice over the past 8 years.

METHOD

Single centre retrospective observational study. Patients included with NSCLC treated with WBRT for radiologically conﬁrmed intracranial (IC) metastatic disease January 2018 to December 2022. OS, baseline tumour/patient characteristics and lines of treatment were recorded from medical records. Data compiled using Excel and statistical analysis using the R language for statistical programming.

RESULTS

62 patients were included, median age 64 (37 -82). 29(46.8%) were Performance status 0-1. 50(80.6%) received 20Gy in 5 fractions, 12(19.4%) received 30Gy in 10 fractions. PDL1 >50% in 14(22.6%), EGFR mutation positive in 10(16.1%), 0(0%) ALK/ROS mutations. 9(14.5%) had received prior radiotherapy(RT) for IC disease (SRS:8, partial brain VMAT:1). 20(32.3%) had IC disease at presentation. Median OS 68 days (95%CI:58-111;range12-777) in keeping with QUARTZ results (median OS 65 days, 95%CI:50- 78). Univariate analyses demonstrated: subsequent systemic therapy (p=0.0022), receiving immunotherapy (p=0.002) and baseline performance status (p=0.0017) associated with improved OS. PDL1 >50%, EGFR mutation positive, RT dose, prior systemic therapy, histological subtype, presence of a targetable mutation, ﬁrst or subsequent IC treatment did not predict survival. The chi-squared test was used to compare differences in the groups with OS>100days(n=22) and OS<100days(n=40). Systemic therapy after WBRT(p=0.0011) and immunotherapy treatment(p=0.0092) were signiﬁcant differences between the cohorts and associated with improved survival.

CONCLUSION

This data supports conclusions of QUARTZ study and it remains diﬃcult to deﬁne a role for WBRT in the management of IC disease in NSCLC. There is a cohort who have better OS, though we believe this may be related to systemic therapy (particularly immunotherapy) exposure.
ARGININE DEPRIVATION SENSITISES GBM CELLS TO RADIATION THROUGH DOWNREGULATION OF DNA REPAIR PATHWAYS
Authors: Perryman Dr Richard , Mussa Ms Hamida , Ghosh Mr Avik , O’Neill Mr Kevin , Syed Dr Nelofer
Published: October 2024

AIMS

Glioblastoma (GBM) is the most aggressive form of brain cancer, and therapy hasn’t changed for almost 20 years. We have previously shown that GBM tumours lack expression of argininosuccinate synthase 1 (ASS1), a key enzyme involved in arginine biosynthesis, due to promoter methylation. This renders them sensitive to systemic arginine depletion using agents such as pegylated arginine deiminase (ADI), allowing us to circumvent the issue of blood-brain-barrier (BBB) penetration. Furthermore, ADI appears to synergise with the DNA damaging agent temozolomide (TMZ), which is routinely used to treat patients with GBM. Here, we aimed to explore whether ADI can synergise with radiotherapy, and determine the mechanisms by which ADI is able to sensitise cells to DNA damaging agents.

METHOD

Primary GBM cell lines were treated with ADI in combination with radiation and underwent analysis using an RNA microarray, Qiagen DNA repair RT2 array, qPCR, western blotting, and proliferation assays in both 2D (SRB) and 3D (spheroid, CCK8) format. We performed orthotopic injection of primary GBM cells into the striatum of athymic nude mice, and randomised them to ADI, radiation, or the combination, plus controls. Tissue was analysed using IHC staining.

RESULTS

In ASS1 methylated primary GBM, treatment with ADI led to a signiﬁcant change in the expression of ~500 genes with a fold change >2. Gene set enrichment analysis indicated that these genes were predominantly involved in cell cycle regulation, which is closely linked to the DNA damage and repair machinery. In another ASS1 methylated primary GBM cell line, an RT2 array revealed speciﬁc downregulation of genes involved in DNA repair. qPCR and western blot validation of these changes showed a signiﬁcant decrease in the expression and protein levels of PARP1 and LIG1 with ADI treatment.

CONCLUSION

Arginine deprivation using ADI leads to a downregulation of PARP1 in GBM cells, sensitising them to radiotherapy.
DYSREGULATION OF M6A RNA METHYLATION, R-LOOPS AND THE DNA DAMAGE RESPONSE IN PAEDIATRIC HIGH-GRADE GLIOMAS
Authors: Walker Ms Lauryn , Radhi Dr Masar , Knight Dr Helen , Smith Dr Stuart
Published: October 2024

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.