bluuebunny/crossref_metadata_2025_split huggingface dataset:
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

Abstract

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

Abstract

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

Abstract

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

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 (&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.
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

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 &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 &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&gt;100days(n=22) and OS&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.
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

Abstract

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 &gt;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

Abstract

AIMS
Paediatric high-grade gliomas (pHGG) are virtually incurable malignant brain tumours. The regulation of R- loops is necessary for ensuring genomic stability. The formation of R-loops at DNA damage sites promotes DNA repair via m6A methylation of the RNA strand. We aimed to investigate whether m6A methylation, R-loop homeostasis and the DNA damage response is dysregulated in pHGG and whether inhibitors, such as ATM inhibitors, could be potential therapeutics for pHGG.

METHOD
To assess the global levels of m6A, R-loops, yH2AX, total ATM and phosphorylated ATM in paediatric HGG cell lines compared to control human astrocytes, immunocytochemistry was initially performed. Immunocyto- chemistry was also performed to assess localisation between m6A and R-loops, R-loops and yH2AX, R-loops and total/phosphorylated ATM, m6A and total/phosphorylated ATM, and total/phosphorylated ATM and yH2AX in pHGG cell lines and human astrocytes. We are also exploring two ATM inhibitors, AZD1390 and AZD0156, and are currently investigating their effects on m6A methylation, R-loops and the DNA damage response in pHGG. As a molecular mechanism involving the tumour suppressor ARID1A was recently discovered that is crucial for maintaining genomic stability, we are also exploring the role of this protein in pHGG. We are also investigating the potential involvement of m6A readers in the pathogenesis of pHGG.

RESULTS
Immunocytochemistry revealed that dysregulation of global m6A, R-loop, yH2AX and ATM occurs in pHGG cell lines as higher levels of expression was generally observed compared to human astrocytes. Moreover, m6A, R-loops, yH2AX and ATM appear to colocalise as high Pearson’s correlation coeﬃcient values were obtained. Experiments exploring ATM inhibitors, ARID1A and m6A readers in pHGG are currently in progress, however the results will be presented.

CONCLUSION
It appears that dysregulation of m6A methylation, R-loop homeostasis and the DNA damage response occurs in pHGG and that molecular mechanisms involving m6A methylation and R-loops could be impaired.
METABOLOMIC PROﬁLING OF ASTROCYTE-GLIOBLASTOMA CELL INTERACTION IN A POSTOPERATIVE TUMOUR MICROENVIRONMENT MODEL
Authors: Kontrimaite Ms Une , McCrorie Dr Phoebe , Porter Mr Harry , Smith Dr Stuart , Kim Dr Dong-Hyun , Rahman Dr Ruman 
Published: October 2024

Abstract

AIMS
Despite multimodal treatment, 9/10 patients diagnosed with isocitrate dehydrogenase wild-type glioblastoma experience tumour recurrence within 5 years. Poor prognosis is in considerable part attributed to the ability of cancer cells to inﬁltrate healthy brain. We hypothesise that metabolic changes underpinning cellular crosstalk between astrocytes and glioblastoma invasive margin cells within a physiologically representative in-vitro tumour microenvironment, may reveal therapeutic targets to prevent or delay glioblastoma recurrence. An initial study aimed to establish baseline metabolomic proﬁles for glioblastoma invasive margin cells and healthy human frontal lobe astrocytes.

METHOD
We used human frontal lobe astrocytes (n=6) and transgenic glioblastoma invasive margin cells tagged with eGFP (n=18) to extract cellular metabolites using biphasic extraction method (methanol:water:chloroform). Liquid Chromatography – Mass Spectrometry (LC-MS) with a ZIC-pHILIC column, coupled to an Orbitrap Mass Spectrometer (Q-Exactive Orbitrap) was used to identify metabolites. Both multivariate and univariate statistical analyses were used to investigate signiﬁcant alterations observed in the metabolic data.

RESULTS
The study uncovered distinct metabolic proﬁles differentiating glioblastoma invasive margin cells from astrocytes. Signiﬁcant alterations were identiﬁed in the alanine, aspartate, and glutamate metabolism pathway, which is required for amino acid synthesis and neurotransmitter balance. Other metabolic pathways include glycine, serine, and threonine metabolism as well as glycerophospholipid metabolism. These preliminary ﬁndings lay the groundwork for further exploration into the cellular interactions in a postoperative tumour microenvironment model and elucidate mechanisms that may contribute to tumour recurrence.

CONCLUSION
Understanding the interactions between cancerous cells and astrocytes is crucial for developing new therapeutic approaches. Our subsequent research will integrate an advanced tumorsphere model composed of a scaffold combining PEGDA hydrogel and an extracellular matrix derived from a decellularized human autopsy brain. This model aims to replicate the postoperative tumour microenvironment more accurately to investigate astrocyte-glioblastoma cell interactions associated with tumour recurrence.
STUDIES ON GLYCOGEN PHOSPHORYLASE AS A POTENTIAL MOLECULAR TARGET IN GLIOBLASTOMA
Authors: Schram Ms Niah , Hayes Dr Joseph , Antia Dr Imeobong , Stasik Dr Izabela 
Published: October 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.
M6A RNA METHYLATION AS A THERAPEUTIC TARGET IN HIGH GRADE GLIOMA
Authors: Radhi Dr Masar , Rowlinson Mr Jonathan , Walker Ms Lauryn , Deacon Dr Simon , Knight Prof Helen , Kim Prof Dong-hyne , Smith Dr Stuart 
Published: October 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.