bluuebunny/crossref_metadata_2025_split huggingface dataset:
DEVELOPMENT OF GLIEXP; A HIGH-THROUGHPUT EX VIVO DRUG SCREENING PLATFORM FOR GLIOBLASTOMA
Authors: Gagg Hannah , Williams Sophie , Rominiyi Ola , Danson Sarah , Collis Spencer , Wells Greg 
Published: September 2023

Abstract

AIMS
With diminishing returns and common clinical failure rates from traditional preclinical and animal-based drug discovery strategies, emphasis is being placed on alternative drug discovery platforms, such as ex vivo approaches. Such approaches represent a departure from both preclinical animal-based models and more traditional clinical-based strategies and aim to address intra/inter-patient variability at an earlier stage of drug discovery. Additionally, they could also offer precise treatment stratification for patients within a week of surgery to direct a tailored treatment course. One tumour group that could significantly benefit from such ex vivo approaches are high-grade gliomas, which exhibit extensive heterogeneity, cellular plasticity and therapy- resistant glioma stem cell (GSC) niches, and where historic preclinical models have failed to generate new therapies leading to 50-year stagnant dire survival rates of around 15 months post-diagnosis.

METHOD
We have developed and optimized a high-throughput ex vivo drug screening platform; GliExP, which maintains GSC populations using immediately dissociated fresh surgical tissue.

RESULTS
As a proof-of-concept for GliExP, we have optimized standard-of-care chemoradiotherapy responses which can correctly predict MGMT status based on Temozolomide sensitivity. We have screened 30+ small molecule therapeutics and preclinical compounds against 18 different patient’s tumours, including multi-region spatial het- erogeneity sampling from several individual tumours. Drug responses are further correlated to transcriptomic and genetic profiles.

CONCLUSIONS
Our data provides a strong basis to build upon GliExP to incorporate combination-based oncology therapeutics in tandem with standard-of-care therapies as an important preclinical model to triage experimental therapeutics for clinical translation, delivering rapid identification of effective treatment strategies for individual gliomas.
M6A RNA MODIfiCATION AND A DRIVER OF CELLULAR PLASTICITY AND INTRA-TUMOURAL HETEROGENEITY IN GLIOBLASTOMA
Authors: Deacon Simon , Radhi Masar , Rowlinson Jonathan , Knight Helen , Kim Dong-Hyun , Smith Stuart 
Published: September 2023

Abstract

AIMS
The immense cellular plasticity of glioblastoma is a barrier to successful therapy. N6-methyladenosine (m6A) methylation of RNA is increasingly recognised as a key driver of cellular differentiation. Crucially, it is a reversible and dynamic modification that alters cellular phenotype via the regulation of mRNA metabolism. We hypothesise that m6A methylation mediates glioblastoma plasticity, regulating transcriptional networks and thus influencing cellular adaptation under different microenvironmental conditions. Our aims are to assess whether total m6A levels and m6A-regulatory gene expression are responsive to change in oxygenation levels and cellular confluency. Furthermore, we aim to characterise the spatial distribution of m6A in 3D cell culture models.

METHOD
We have investigated m6A levels in patient-derived cell lines from the invasive region of adult glioblastoma. Mass spectrometry and anti-m6A immunofluorescence confocal microscopy was used to quantify total m6A levels. We have compared total m6A levels in normoxic versus hypoxic culture conditions and in low versus high confluency 2D cell culture, including cell migration during a scratch assay. We have measured expression levels of key m6A-regulatory genes using rtPCR.

RESULTS
Total m6A levels and regulatory gene expression were not significantly different in normoxic versus hypoxic conditions. However, m6A levels were decreased in high confluency versus low confluency conditions. We will further characterise the spatial distribution of m6A in 3D cell culture models using immunofluorescence confocal microscopy.

CONCLUSIONS
We demonstrate a degree of variability in m6A levels in response to micro-environmental conditions, such as cellular confluency. These findings indicate a role for m6A in tumoral heterogeneity and cellular adaptation.
QUANTIFYING THE NEED FOR A BEST SUPPORTIVE CARE CLINIC
Authors: Al-Munaer Marwan , Holman Sam , Wilby Emma , Brodbelt Andrew 
Published: September 2023

Abstract

AIMS
Patients newly diagnosed with high-grade primary brain tumour and poor performance status discussed in the Multidisciplinary team (MDT) may not be felt to benefit from active treatment and their uptake in clinic is poor. We aimed to explore the volume of patients not being treated in our region, their survival rates, and the reasons for the limited requests for clinical review.

METHOD
Data was obtained from medical records of identified patients from April 2021 to April 2022. This included demographics, presentation, diagnosis date, clinic visits, and date of death.

RESULTS
78 patients were referred for suspected high-grade primary brain tumours and selected for best supportive care by the MDT. Only 73 (64% male) were diagnosed radiologically with a high-grade primary tumour and the rest were excluded.
Median age was 79 years (range 74 – 84). The commonest presenting symptoms were confusion (53%), falls (29%), and dysphasia (25%). Median survival time after diagnostic scan was 67 days (range 34 – 107). Survival rates at 1, 3, 6, and 12 months were 82%, 33%, 8%, and 1%, respectively.
28 patients (38%) were seen in clinic. 13 had been provided with the option to attend by letter. The median duration from referral to clinic was 9 days (range 7 – 14).

CONCLUSIONS
Only a third of non-treatable patients with newly diagnosed high-grade primary brain tumours are being seen in the clinic. Communication between the MDT and the patient offering this service is poor. Longer term survivors suggest an incorrect diagnosis who may benefit from further review.
ALTERNATIVE LENGTHENING OF TELOMERES (ALT) IS SEEN IN A PROPORTION OF OLIGODENDROGLIOMAS AND IT IS ASSOCIATED WITH A WORSE PROGNOSIS
Authors: Ng Ho Keung , Li Kay K K , Shi Zhi-Feng 
Published: September 2023

Abstract

AIMS
Oligodendrogliomas are well known to be mutated for TERTp, which is enigmatically a diagnostic criteria for molecular glioblastoma by WHO2021. It is generally believed that gliomas that are TERTp negative maintain their telomere lengths by means of Alternative Lengthening of Telomeres (ALT) which is usually reflected by mutations of ATRX. Oligodendrogliomas are known to be non-mutated for ATRX unlike astrocytomas. TERT re-arrangement has also been shown to be an alternative way of TERT activation.

METHOD
We evaluated 112 oligodendrogliomas of both Grades 2 (n=87) and 3 (n=25) (all IDH-mutant, 1p19q co-deleted and 93% TERTp mutated by Sanger sequencing) by FISH for ALT, MYC, PDGFRA, EGFR , CDKN2A HD, and +7/-10, the latter events being associated with malignant transformation in low-grade gliomas.

RESULTS
40 cases (35.7%) showed ALT and ALT positivity was associated with worse PFS (p=.024) and remained an in- dependent prognosticator in multivariate analysis. Amplification was found in MYC (6.3%), PDGFRA (10.7%), EGFR (0.8%), CDKN2A HD (6.25%) and +7/-10 (0%) and they had no prognostic significance, likely due to low occurrences. ALT was found to be associated with MYC amplification (p=.004). ALT-positive cases were further evaluated by FISH for TERT re-arrangement, and targeted panel DNA sequencing. CIC mutation (62.1%) was as- sociated with shorter OS in Grade 2 tumours (p=.023) and was the only significant finding from panel sequencing. TERT re-arrangement was only seen in 10.7% and had no prognostic significance.

CONCLUSIONS
ALT is seen in a proportion of oligodendrogliomas in spite of TERTp mutations and is associated with a worse prognosis.
ROLE OF THE TRANSCRIPTIONAL ELONGATION FACTOR SPT6 AS A MODULATOR OF RAD52 IN GLIOBLASTOMA TUMOURS
Authors: Aguilar-Morante Diana , Hamerlik Petra 
Published: September 2023

Abstract

AIMS
Glioblastoma (GBM) is the deadliest and most common primary brain tumour with median survival rates of less than two years. Inter- and intra-tumoral heterogeneity, cellular plasticity and de-regulated signalling pathways are plausible causes of resistance to existent therapies in GBM. As current therapies offer only limited survival benefits, the identification and validation of new approaches in glioblastoma management is of the highest importance

METHOD
• In silico analysis, RNA sequencing analysis after SPT6 depletion and analysis of R-loop formation after RAD52 depletion.
• Patient-Derived Xenografts and primary human glioblastoma cells

RESULTS
Our recent findings show significantly higher RAD52 mRNA levels in malignant gliomas compared to normal brain, where its levels negatively correlate with GBM patient survival. Experimental data show that the inhibition of RAD52 by 6-Hydroxy-DL-Dopa (6OH-Dopa) impairs GBM cell survival due to increased replication- transcription collisions leading to the accumulation of DNA damage. Transcriptional elongation factor SPT6 has previously been named an essential gene in GBM. We find that loss of SPT6 decreases the expression of HR proteins BRCA1, RAD51 and RAD52. RAD52 binds to single-strand DNA (ssDNA), plays a crucial role in most HR events, and restarts collapsed replication forks in response to oncogene-induced replication stress. Further investigation into the role of RAD52 showed depletion of RAD52 results in an increase of R-loops increasing genomic instability and DNA damage.

CONCLUSIONS
Based on our findings, we hypothesize that SPT6 modulates RAD52, which acts as a sensor of R-loops and plays a crucial role in R-loops’ resolution.
DEVELOPING 3D PRINTING MODELS OF BRAIN TUMOURS
Authors: Ballesteros-Cillero Rafael , Valli Jessica , Melchels Ferry P W , Leslie Nicholas R 
Published: September 2023

Abstract

AIMS
To succeed in clinical trials for glioblastoma we need in vitro models capable of more faithfully replicating dis- ease biology and more accurately predicting patient drug responses. To this end, new bioprinting technologies have the potential to biofabricate clinically relevant biomimetic tissues which can accelerate drug discovery and additionally serve as a platform for personalized medicine.

METHOD
We evaluated the effect of individual biomaterials and combinations of biomaterials, including decellularised pig brain extracellular matrix (dECM), fibrin, gelatin-methacryloyl (GelMA), hyaluronic acid-methacrylate (HAMA), Matrigel and alginate, on the proliferation and invasion of aggressive brain cancer cells (U87) in vitro. Cell viability was assessed using propidium iodide. Invasiveness was studied employing confocal microscopy. Data generated from Z-stacks was analysed using ImageJ to determine the size and circularity of cells.

RESULTS
Although Matrigel supports rapid cell proliferation and invasion, it has mechanical properties unsuited to bio- printing. In contrast, HAMA displays a pronounced shear-thinning behaviour and rapid controllable photo- crosslinking. Combinations of HAMA-fibrin provided results comparable to those seen with Matrigel or HAMA- Matrigel. However, high levels of crosslinking affected these biomaterial mixtures, resulting in a decreased ability of cells to grow and spread.

CONCLUSIONS
Initial results indicate that the addition of fibrin to HAMA promoted the growth and spreading of U87 cells. In further work, we aim to improve our printed constructs by including porcine brain dECM, microglia and recently established cell lines from paediatric patients. The project will test whether these bioprinted models can provide drug testing data with closer results to human disease than current, simpler alternatives.
M6A RNA METHYLATION AS A THERAPEUTIC TARGET IN HIGH GRADE GLIOMA
Authors: Radhi Masar , Rowlinson Jonathan , Halliday Nigel , Kim Dong-Hyun , Deacon Simon , Knight Helen , Smith Stuart 
Published: September 2023

Abstract

AIMS
High grade glioma (HGG) is a devastating brain cancer that has poor prognostic outcomes. N6-methyladenosine (m6A) is the most prevalent mRNA modification. It is co transcriptionally installed by a methyltransferase com- plex composed of METTL3, METTL14 and WTAP. The modification is reversible and erased by FTO and ALKBH5. Here we sought to investigate the functional importance of m6A RNA methylation in the pathogenesis and drug resistance of HGG.

METHOD
To identify the expression of key m6A effectors, RT-qPCR and immunofluorescence were undertaken . The expression of these effectors was modified using siRNA knockdown and small molecules to identify their effects on key transcription factors. The effect of modification on the ability of the cells to form neurospheres 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 m6A level was achieved by developing methylated RNA immunoprecipitation qPCR and LC-MS/MS techniques.

RESULTS
We found that key m6A RNA methylation effectors are highly expressed in HGG cell lines and tissues. LC-MS/MS and immunofluorescence showed an increase in the level of m6A. Genetic and chemical inhibition of METTL3, FTO and WTAP affects the expression of key transcription factors in self-renewal, cell cycle regulation and tumorigenesis. Depletion of these effectors reduces the ability of cells to form neurospheres, increases apoptosis and suppresses invasion.

CONCLUSIONS
These findings suggest the role of RNA methylation in the pathogenesis of HGG and could open a new avenue for developing effective treatment strategies.
BEHAVIOURAL TESTING OF SENSORI-MOTOR FUNCTION IN AWAKE CRANIOTOMY; A LITERATURE REVIEW
Authors: Cooper Sara , Sinha Rohit , Stephenson Elisa , Ewan Seonaid , Mathew Ryan 
Published: September 2023

Abstract

AIMS
Evaluate how sensori-motor (SM) testing is performed in awake craniotomy (AC) in the literature. Compare SM testing methods and their related outcomes.

METHOD
Literature search from 1995- 2019 in OVID Medline, Embase, PsychInfo, CINAHL and Web Of Science using all permutations of MESH terms “brain tumour” “awake craniotomy” “sensory” “motor” “testing”. 11671 articles were screened and 28 included based on suffcient detail of the tasks and intraoperative processes that would allow for evaluation.

RESULTS
SM testing is used to preserve function in AC when tumours involve or neighbour eloquent cortical structures sub serving movement. It is also used to identify subcortical tracts such as corticospinal and frontostriatal, as well as and the negative motor network. There is broad variation in how SM testing is conducted in different centres, with no standardisation. The most common strategies were:
•Simple gross movement of contralateral limbs (n= 17) e.g., hand open/closing.
•Functional movement (n=5) e.g., grasping object
•Standardised tests (n= 3) including the Hand Manipulation Task and Sensory-Motor Profile awake
•Patient at rest observed for positive motor responses or patient reported sensory changes (n= 2).

CONCLUSIONS
Multiple different intra-operative strategies exist for the preservation of sensori-motor function in awake craniotomy. Benefits of the over-arching AC strategy for SM preservation is frequently demonstrated. However, comparison and optimisation between strategies is hindered by heterogenous study objectives and outcomes reported. A core testing and outcome set may be useful to standardise, optimise and compare so that maximal patient benefit and functional preservation can be achieved.
INVESTIGATING THE ROLE OF LIPID DROPLETS IN MEDULLOBLASTOMA TUMOURS
Authors: Cotton Kian , Niklison-Chirou Maria Victoria 
Published: September 2023

Abstract

AIMS
Medulloblastomas (MBs) are the most common paediatric brain tumour which originates in the cerebellum. It is classified into four subgroups named: sonic hedgehog, wingless, Group 3 and Group 4. Current treatment for MBs includes surgery, followed by high levels of chemotherapy and radiotherapy. This treatment regime is highly aggressive for the developing brain and produces several significant side effects. Therefore, research into new treatments or improving those already available is crucial to improving patients of life.

METHOD
It is well-documented that cancer cells have the ability to reprogram their metabolism to support rapid proliferation. These metabolic adaptations will provide the cells with the ability to use alternative sources of energy such as glutamine or lipids. Lipid Droplets (LDs) are highly dynamic cytoplasmic organelles that, under homeostatic conditions, are key in the storage of excess lipids within the cell to provide a buffer against lipotoxicity. Recent research has also begun to describe LDs as more than simple storage molecules but instead as function- ally active organelles. Increases in LD formation have been reported in several cancers such as Glioblastoma

RESULTS
In this study, we observed that aggressive MB cells have significantly more LDs when compared to non- aggressive MB cells. Further, oleic acid stimulation induced LD formation in MB cells. Finally, in silico gene expression analysis was performed to evaluate the expression of multiple components of the de novo lipid synthesis pathway in medulloblastoma.

CONCLUSIONS
Our work demonstrates that aggressive MB cells overexpress LDs and therefore identifies them as a possible novel target for MB treatment.
INVESTIGATING LOCAL RECURRENCE IN SECONDARY BRAIN TUMOURS
Authors: Suppree Joshua , Jenkinson Michael D , Zakaria Rasheed 
Published: September 2023

Abstract

AIMS
Brain metastases result in considerable morbidity and mortality. Modern treatment strategies are increasingly omitting radiation therapy after surgical resection of these tumours. We investigated the rate of local recurrence in patients who had undergone surgery for brain metastasis in a single-institution study and compared it to the literature through a comprehensive meta-analysis.

METHOD
Patients undergoing surgery for a brain metastasis were identified and their records reviewed. Data on treatment and outcome was extracted and analysed. A systematic-review and meta-analysis, registered in PROSPERO (CRD42023391637), was conducted following the PRISMA statement.

RESULTS
The single-institution study comprised 358 cases, 40% (n=142) male and 60% (n=216) being female with a median age at surgery of 60 years (range 23-85 years). The most frequent primary cancer was lung cancer affecting 35% (n=126), followed by breast cancer (24%, n=87) and malignant melanoma (13%, n=47). Among the cases 142 underwent surgery without any adjuvant radiation. The median freedom from local recurrence at 12-months for this population was 38.3% (95% CI 13.4, 44.6). The meta-analysis encompassed three studies, with a total of 170 cases. Freedom from local recurrence at 12-months was found to be 42.4% (95% CI 34.9 - 50.1) following surgery alone.

CONCLUSIONS
We define for the first time the rates of local recurrence following surgery alone for brain metastases with a large single-centre series and a meta-analysis. As radiation is increasingly being omitted in these cases, caution should be exercised, and more investigation of the factors associated with local recurrence after surgery and how they be modified are required.
INVESTIGATING NOTCH1 PATHWAY COMPONENTS IN HISTONE 3.3 MUTANT PAEDIATRIC HIGH-GRADE GLIOMA
Authors: Emayakumaran Emayanthi , Lourdusamy Anbarasu , Layfield Robert , Haque Farhana 
Published: September 2023

Abstract

AIMS
High-grade gliomas (HGG) in adults and children (paediatric high-grade glioma, pHGG) differ in molecular characteristics, despite having the same morphological features. Although pHGG has a low incidence, it contributes to 40% of all brain tumour death in childhood. Histone 3 (H3) mutation is a unique feature of pHGG. The understanding the pathophysiology of these tumour are important to develop more targeted therapy for these tumours.
Our previous research has identified, increased activation of the Notch1 pathway in H3 mutated tumours. We have now explored the relationship between the Notch1 pathway components to H3 mutated pHGG. We hypothesised that The Notch1 pathway components (NICD1, HES1, HES5) are dysregulated in the Histone 3 mutant tumours in comparison to H3 wild type tumours.

METHOD
Protein expression analyses, immunofluorescence and microscopy and western blot experiments were per- formed, to assess the localization and expression of Notch1 pathway components (NICD1, HES1, HES5), using cultures H3.3 wildtype and H3.3 mutant cell lines.

RESULTS
The results show that Notch1 pathway expression differ between H3 mutant and H3 wild type tumours.

CONCLUSIONS
These results suggest that components of Notch1 pathway (HES1/HES5) are dysregulated, which may be driving tumour growth in this H3 mutated tumour. Therefore, we can explore Notch1 Pathway components as targeted therapeutic options.
PROCARBAZINE, LOMUSTINE, VINCRISTINE FOR THE SECOND LINE TREATMENT OF GLIOBLASTOMA MULTIFORME
Authors: McCormick Cecilia , Gkogkou Pinelopi , Ho Andrew 
Published: September 2023

Abstract

AIMS
Most patients with glioblastoma recur despite surgery and post-operative chemoradiation. The optimal second line chemotherapy regime is not known. The aim of this study was to evaluate PCV chemotherapy given as second line treatment for glioblastoma.

METHOD
Data was collected for all patients with glioblastoma treated with second line PCV between 2009 and 2021 at the Norfolk and Norwich University Hospital. All underwent surgical resection either complete or incomplete resection followed by adjuvant radiotherapy and chemotherapy according to the Stupp or Perry A protocol. All received palliative PCV chemotherapy at recurrence. The Response Assessment in Neuro-Oncology (RANO) criteria were used to assess response to palliative PCV chemotherapy. Patient and disease related characteristics were described. Disease free survival from recurrence to death and survival outcomes were estimated using Kaplan-Meir method.

RESULTS
70 patients were evaluated. Overall survival from surgery to death was 290 days (95%CI 245 to 318)
Survival from disease progression to death was 182 days (95%CI 131 to 218). Subgroup analysis was performed for patients &lt;65years and &gt;65years. For patients &lt;65 years N=51, median overall survival was 199 days (95%CI 142 to 353) p value 0.1246. For those &gt;65 years N=16, median overall survival was 130.5 days (95%CI 93 to 227)

CONCLUSIONS
Using PCV as a second line chemotherapy for patients with glioblastoma is a reasonable approach. It is well tolerated and provides benefit for patients &lt;65 years as well as older patients.