tbuckley/cabot-search huggingface dataset:
WHOLE BRAIN RADIOTHERAPY FOR INTRACRANIAL METASTATIC DISEASE IN NON-SMALL CELL LUNG CANCER: THE WESSEX EXPERIENCE
Authors: Dr Eleanor Starkey, Dr Adityanarayan Bhatnagar, Dr Andrew Bates, Dr Paul Fenton, Dr Victoria Wood, Ms Nelima Begum, Dr Sean Main
Journal: Neuro-Oncology (2024)

Abstract 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 &amp;gt;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 &amp;gt;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&amp;gt;100days(n=22) and OS&amp;lt;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.
FROM VEIN TO BRAIN: ENHANCING TRAﬃCKING OF CAR T-CELLS IN DIFFUSE MIDLINE GLIOMA
Authors: Dr Louise Steele Saukila, Dr Alastair Hotblack, Dr Christina Burke, Ms Carmen Rodriguez, Dr Matteo Righi, Dr Tammy Kalber, Dr Antonia De-Cola, Mr Michael McNicholas, Dr Manav Pathania, Dr Martin Pule, Dr Karin Straathof
Journal: Neuro-Oncology (2024)

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.
M6A RNA METHYLATION AS A THERAPEUTIC TARGET IN HIGH GRADE GLIOMA
Authors: Dr Masar Radhi, Mr Jonathan Rowlinson, Ms Lauryn Walker, Dr Simon Deacon, Prof Helen Knight, Prof Dong-hyne Kim, Dr Stuart Smith
Journal: Neuro-Oncology (2024)

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.
DIFFERENTIAL CELL SURFACE PROTEOMICS OF FLOTILLIN-1, HMGB1 AND AMINOPEPTIDASE N IN H3 MUTATED PAEDIATRIC HIGH-GRADE GLIOMAS
Authors: Ms Hannah Fairley, Ms Chloe Walugembe, Mr Joshua Sulman, Dr Farhana Haque
Journal: Neuro-Oncology (2024)

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.
OCCUPATIONAL THERAPY PRE-ASSESSMENT OF LOW GRADE GLIOMA PATIENT WITH COMPLEX NEEDS ALMOST HALVES POST-OPERATIVE LENGTH OF STAY
Authors: Ms Claire Laycock, Ms Sunita Sokhal, Mr Ryan Mathew
Journal: Neuro-Oncology (2024)

Abstract AIMS Due to tumour eloquence, patients with low grade gliomas (LGGs) often have complex Occupational Therapy (OT) needs post-operatively. This need can impact length of stay and OT resource. Limited studies have focused on pre-assessment for this patient group. To address these challenges, we aimed to identify the impact of a newly-introduced Occupational Therapy (OT) pre-surgical assessment service for low grade glioma patients. METHOD Patients (n=16, prospective, consecutive, Jan-Jul 2023) planned for LGG surgery at Leeds Teaching Hospitals NHS Trust were offered an OT pre-assessment. Mixed methods analysis. Assessments and data collected included existing needs and deﬁcits, length of stay, and OT sessions (number/contacts, time spent). Comparison was made to patients (n=16, retrospective, consecutive, Jun-Dec 2022) planned for LGG surgery who had not received this service. Complexity was deﬁned using standardised, clinically-accepted factors (social, cognitive and/or physical deﬁcits). RESULTS Patients with complex needs (4 in each group) beneﬁtted signiﬁcantly from OT pre-assessment with reductions in: length of stay (11.9 v 6.5 days, 45.4% decrease), median time spent (134 v 95 mins, 29.1% decrease), median number of contacts (5.4 v 3.4, 37.0% decrease). Patient, surgeon, CNS and other AHP feedback was overwhelm-ingly positive. CONCLUSION Patients with complex needs undergoing surgery for low grade gliomas signiﬁcantly beneﬁt from pre- assessment; reducing length of stay and liberating occupational therapy resource. This also reduces costs and increases productivity/eﬃciency. Further work in more LGG patients, other brain tumour patients, and the identiﬁcation of key pre-assessment variables and interventions will guide the development of the service.
THE IMPACT OF THE FRONTAL ASLANT TRACT ON POST-OPERATIVE OUTCOMES IN GLIOMA SURGERY
Authors: Mr Kapil Rajwani, Mr Jose Lavrador, Mr Azharul Khan, Dr Qusai Al Banna, Mr Asfand Mirza, Mr Jahard Aliaga-Arias, Mr Ali Elhag, Mr Francesco Marchi, Mr Jonathan Shapey, Mr Ranjeev Bhangoo, Prof Keyoumars Ashkan, Mr Richard Gullan, Mr Francesco Vergani
Journal: Neuro-Oncology (2024)

Abstract 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 (&amp;gt;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.
INTRAOPERATIVE MRI FOR BRAIN TUMOURS: A 15-YEAR RETROSPECTIVE REVIEW AT A SINGLE AUSTRALIAN INSTITUTION
Authors: Dr Daniel Madani, Dr Krishna Muralidharan, Dr Nick Chang, Mr Patrick Tang, Dr Johnny Wong
Journal: Neuro-Oncology (2024)

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.
THE EPIGENETIC REGULATORY PROTEIN CBX2 AS A NOVEL THERAPEUTIC TARGET FOR GLIOBLASTOMA
Authors: Ms Danielle Marsh, Prof John Greenman, Dr Mark Wade
Journal: Neuro-Oncology (2024)

Abstract 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.
APPLICATION OF IN VITRO MODELS: INCORPORATING BLOOD BRAIN BARRIER MODELS WITH GLIOMA, AND THE USE OF MICROﬂUIDICS
Authors: Ms Eleanor Taylor-Chilton, Dr Lydia Baldwin, Dr Lucy Stead, Prof John Greenman, Dr Mark Wade, Dr Tamer Rabie, Dr Jane Alder
Journal: Neuro-Oncology (2024)

Abstract AIMS The blood brain barrier (BBB) presents one of the main obstacles for the development of novel glioblastoma treatments. The various transporters and enzymes within the BBB reduce the disposition of novel therapies into the brain. There is a need for an in vitro BBB-glioma model with proven eﬃcacy of predicting CNS delivery at earlier stages in the drug discovery pipeline. METHOD A BBB-glioma lab-on-a-chip model could enable rapid identiﬁcation of novel therapies that can cross the BBB at earlier stages of the drug development process, before going to clinical trials. The application of microﬂuidic technology allows for the detection of soluble tissue-derived factors and the assessment of the levels of soluble markers released from malignant tissue. Adding ‘ﬂow’ to the model, is more reﬂective of the dynamic ﬂow present at the BBB. RESULTS Validation studies of the current BBB model and converting this model over to a lab-on-a-chip model have shown the presence of an endothelial cell barrier and the tight junction proteins Zo-1 and Claudin-1, through western blotting, immunocytochemistry, and permeability experiments using varying molecular weights of FitC-dextran. These validation studies have been completed using brain endothelial monolayers on polycarbonate transwell inserts, with the immortalised cell line HCMEC/d3, and the short-term culture cells HBMEC. Also, validation studies using a co-culture of endothelial cells, astrocytes and pericytes have begun, using the same techniques to show a barrier has formed, on both the current BBB model and the lab-on-a-chip model. CONCLUSION The aim of this study is to combine the use of an all-human BBB-glioma model with lab-on-a-chip technology to produce a high throughput model to test novel therapies at an earlier stage. The lab-on-a-chip model will be validated for the purpose of measuring permeability and drug response and to investigate the expression of novel targets verses expression in conventional models.
STUDIES ON GLYCOGEN PHOSPHORYLASE AS A POTENTIAL MOLECULAR TARGET IN GLIOBLASTOMA
Authors: Ms Niah Schram, Dr Joseph Hayes, Dr Imeobong Antia, Dr Izabela Stasik
Journal: Neuro-Oncology (2024)

Abstract AIMS Glioblastoma (GB) is one of the most aggressive malignancies of the brain and spinal cord. Current standard of care has remained surgical resection of the tumour, followed by radiotherapy and/or chemotherapy. Crucial challenges in the effective treatment of GB include resistance to the main chemotherapeutic, temozolomide. Prognosis is poor with median survival remaining at 14.6 months. A now established hallmark of cancer is the ability of malignant cells to “modify, or reprogram, cellular metabolism”, such as glycolysis upregulation. This hallmark could suggest the potential of targeting a speciﬁc metabolic component within glycolysis, thus reduce energy production, resulting in possible apoptosis/suppressed proliferation. Glycogen phosphorylase (GP), an enzyme in glyconeolysis, could be inhibited for a therapeutic effect in GB. In this project, GP levels across the three different isoforms (PYGB, PYGL, and PYGM) and the effect of the GP inhibitor CP-91149 was investigated across three different GB cell lines. METHOD Cell viability of T98G, U251-MG, and U87-MG glioblastoma cell lines and SVGp19 human fetal glial cell line was measured after administration of CP-91149 for 24, 48, and 72 hours. Migration of T98G and U251 cells following treatment with CP-91149 was measured in a wound healing assay. Visualisation of the three GP isoforms was achieved through western blot and conﬁrmed by immunocytochemistry. Flow cytometry was utilized to expand analysis on GP inhibition through assessing cell cycle before and after treatments. RESULTS Results showed CP-91149 had a signiﬁcant dose-dependent effect in vitro on all cell lines. Inhibition of GP resulted in reduced cell migration, also in a dose-dependent manner. Western blot and immunocytochemistry analysis showed PYGL is the most predominant isoform of GP. CONCLUSION Future work in this project will aim to silence expression of GP in GB cell lines in order to compare the results with that yielded from cells treated with the GP inhibitor CP-91149.