bluuebunny/crossref_metadata_2025 huggingface dataset:
P08.08.B Long-term survival and health-related quality of life in meningioma patients: a mixed-methods systematic review
Authors: Frances S , Murray L , Wright J , Velikova G , Boele F
Published: September 2022

Background

Meningiomas account for approximately 36% of all primary brain tumours. These tumours have a relatively favourable prognosis with approximately 92% of meningioma patients surviving >5 years after diagnosis. Yet, patients report high disease burden and survivorship issues even years after treatment, including cognitive issues and anxiety/depression affecting health-related quality of life (HRQOL). We aimed to systematically review the literature and synthesise evidence on HRQOL in meningioma patients across long-term survival, defined as >2 years post diagnosis.

Material and Methods

Systematic literature searches were carried out using Medline, EMBASE, CINAHL, PsycINFO, and Web of Science Core Collection up to November 2021. Two independent assessors carried out screening. Any published, peer reviewed article with primary quantitative or qualitative data covering the physical, mental, and/or social aspects of HRQOL were included. Quality was assessed using the Mixed Methods Appraisal Tool, before a data-based convergent synthesis design was used to interpret findings.

Results

The search returned 2252 articles. Following screening 12 papers were included in the review. The majority of studies used quantitative methods, with only one study reporting mixed-methodology. Two of these were articles from the same study. Most used cross-sectional assessments (n =10) with two reporting longitudinal assessments (n = 2). Included meningioma patient samples (n=2990 in total) represented a wide range of survival lengths (range: 24 months - 10 years). HRQOL was impacted predominantly through physical and mental issues, including persevering symptoms (e.g. headaches, fatigue, vision problems) and emotional difficulties (e.g. poorer mental and general health perceptions). Returning to work proved difficult for patients due to cognitive and social deficits (e.g. issues with concentration, procrastination and isolation). Factors including age and comorbidities, but also treatment type appears to affect patients’ satisfaction with their HRQOL.

Conclusion

While overall prognosis following a meningioma diagnosis is good, meningioma patients’ HRQOL appears affected throughout long-term survivorship. Findings from this review could be beneficial in addressing the HRQOL and supportive-care needs of patients across long-term survivorship.
P08.11.A A subgroup analysis of utilities from European patients using tumor treating fields for glioblastoma
Authors: Palmer J D , Proescholdt C , Chavez G , Furnback W
Published: September 2022

Background

Recently a large-scale, international survey was undertaken studying real-world Health-Related Quality of Life (HRQoL) outcomes for patients using Tumor Treating Fields (TTFields) for treatment of glioblastoma (GBM). It is the largest published HRQoL study in GBM to date (Palmer et al. 2021). The study issued the EuroQol 5-Dimension (EQ-5D) survey to 2,815 patients using TTFields therapy for GBM in the US (2,182 patients) and Europe (633 patients).

Material and Methods

We analyzed EQ-5D results reported from European patients using TTFields for GBM in the above study. We used a EuroQol-endorsed model (Andrade et al. 2020) to estimate health state utilities from these EQ-5D responses for application to the French population. A health state utility is a number that quantifies patient HRQoL, where 0 corresponds to dead, 1 corresponds to perfect health, and numbers in between 0 and 1 correspond to intermediate HRQoL. We additionally considered data on demographics, disease characteristics, and treatment details.

Results

We report final analysis of the 327 EQ-5D responses received from European patients, representing a response rate of 51.7% i.e. the majority of European patients surveyed. The median/mean age was 57.5/56 years. 36.5% of patients were female and 63.5% male. 64.5% of patients had not experienced disease progression while 35.5% were post-progression. 44% of patients reported no additional therapies were being used in addition to TTFields while 56% reported the current use of other therapies. The median/mean time-since-diagnosis was 16/21.4 months and the median/mean time-using-TTFields therapy was 10/14.3 months. The overall median/mean utility estimates were .871/.784. The median/mean utility estimates for patients who had not experienced disease progression were .901/.841, while those for patients who had experienced disease progression were .833/.701. The median/mean utilities for patients under 65 years of age were .872/.778 while for patients at or over 65 they were .855/.791. Univariate regression analyses show that greater than 15 months since diagnosis is correlated with improved utilities (p=.033). These analyses also show that greater than 10 months using TTFields therapy is correlated with improved utilities (p=.005). Multivariate analysis controlling for age, gender, disease progression, other therapies, and time-since-diagnosis still shows that longer time using TTFields is correlated with improved utilities (p=.004).

Conclusion

The high survey response rate and relatively high utility estimates for both progressed and non-progressed patients as well as patients both over and under 65 years of age indicates that many European patients with GBM using TTFields can expect good QoL. Results indicate that utilities for European patients with GBM are likely to improve with longer time using TTFields therapy.
P08.12.B Conceptual development of an intensive supervised exercise program for brain tumor patients: summary of clinical experience
Authors: Jost J , Brandt R , Altuner U , Müther M , Stummer W , Völker K , Wiewrodt R , Wiewrodt D
Published: September 2022

Background

Several studies have identified beneficial effects of physical activity on reducing cancer-related symptoms. However, the number of such studies in neuro-oncology is limited, and even today, brain tumor patients are often advised against physically strenuous exercise. Here we summarize our experience with an individualized training program in brain tumor patients over a period of more than 8 years.

Material and Methods

Patients with primary brain tumors were invited to participate in the individual training program as part of the psycho-oncological consultation. If interested and free of major comorbidity, a professional sport scientist/diploma coach individualized two 60-minute sessions/week adapted to the patients’ respective symptoms and trained them on a 1:1-basis. One session consisted of bicycle ergometry at an average workload of 75% of the max. age-adjusted heart rate (up to 95% at peak; continuous monitoring throughout entire session), the other session was progressive whole-body resistance training based on 10 separate, but standardized exercise tasks using the university’s training facilities. Both training sessions were complimented by challenging elements to improve coordination. Exercise tasks were adapted as needed, and exercise levels were enhanced over time, if possible.

Results

From 2011 to 2019, 45 patients (19 women, 26 men) aged 20-76 years (mean 49) with different tumor types (65% high-grade gliomas, 22% low-grade gliomas, 13% other) participated in the program. The majority of patients started the program following concurrent radio-chemotherapy and in parallel with adjuvant systemic therapy. On average, 41 training sessions were performed. No training-related adverse events (e.g., falls, head pain, etc.) occurred during the entire period. In a total of 1828 training sessions, two minor epileptic seizures occurred (1 speech arrest; 1 simple focal seizure, left hand affected). Both patients were familiar with the respective type of seizure before entering the program and training could be continued immediately, with reduced intensity. Seizures did not reoccur during subsequent training sessions.

Conclusion

This supervised intensive physical training program with submaximal exertion zones was feasible, safe, and highly rated by all participants. Based on these experiences and the reported well-being of the patients, we launched a prospective oligocentric study to objectify the improvements in physical performance and quality of life in patients with glioblastoma (ClinicalTrials.gov Identifier: NCT05015543).
P08.14.B Addressing and implementing coping strategies from pre-surgery can reduce anxiety and depression in the brain tumour population
Authors: hurwitz v , Lavrador J p , La J ,   Kings neuro-oncology, Brazil L , Al-Salihi O , Swampillai A , Bhangoo R , Vergani F , Ashkan K , Chia K , Hedges S , Suarez A , Kostick E , Robinson C
Published: September 2022

Background

Anxiety and depression is prevalent in oncology patients and disproportionately high in neuro-oncology. The screening tools are most frequently used when signs of depression are identified later in the patients pathway. The cognitive impact of a brain tumour means coping mechanisms are harder to integrate and retain. Anti-depressants and their impact on a brain tumour have long since been debated with no clear conclusion. Implementing a programme to address the anticipatory diagnosis of anxiety and depression from first contact potentially allows patients to discuss their mental health needs more openly.

Material and Methods

Patients are introduced to the concept of health related anxiety and depression following their first pre-assessment appointment, A combination of techniques are employed through different mediums including baseline assessment with EORTC QOL screening, MoCa assessment, CBT techniques, YouTube tutorials via the neuro-oncology cns channel and one to one sessions over six to nine sessions.

Results

Patients perceived quality of life is higher following completion of the programme. 75% of participants were able to identify coping strategies they had learnt and how and when to use them. Oncologists reported patients were better prepared for their results and initial oncology consenting appointment. High anxiety levels caused one participant to self-discharge prior to their surgery. On completion of four sessions they returned to the hospital and underwent awake craniotomy.

Conclusion

Identifying and implementing coping strategies at the earliest opportunity improves patients quality of life whilst living with a brain tumour. Anxiety and depression being discussed and the risk of developing either or both of these conditions allows for more effective prevention, awareness and diagnosis. There is the potential to further this work by looking at the impact on carers of early detection of patients anxiety and depression
P09.01.A Speak up - a film project about living with a brain tumor
Authors: Piil K , Mathiasen M , Raffnsøe I , Vollmer V , Risgaard K
Published: September 2022

Background

When a person is diagnosed with a brain tumor everyday life change completely. Patients experience severe and fluctuating symptoms and the relatives often become informal caregivers. Together they seek to manage the symptoms and maintain quality in daily living. Especially, cognitive impairments are difficult to manage and often associated with stigma. The purpose of this project was to create awareness and share knowledge about the challenges experienced by those living with a brain tumor and their informal caregivers. The creation of the films was chosen to be an appropriate medium to reach our targeted public population. Even though film cuts are time-consuming with several costs to produce, they have a beneficial perspective. Among the advantages of this media is that film cuts are easy to access and understand, simple to share on social media, and addresses universal and population-specific problems and concerns making the audience wide ranging. Thereby, the film cuts may help prevent prejudices and misunderstandings from arising, and also seek to provide patients with a language and courage to talk about difficult sensitive issues.The Danish Brain Tumor Association (Hjernetumorforeningen) has produced nine short films about being diagnosed with and living with a brain tumor diagnosis. We intend that the film cuts will support and inform patients, relatives/family, friends, and professionals to better understand the complex situations experienced by brain tumor patients.

Material and Methods

A screenwriter from The Ophelia School of Acting attended several network meetings held by the ‘Hjernetumorforeningen’ and interviewed patients, relatives, health care professionals and board members of ‘Hjernetumorforeningen’ to write the manuscript of each film. Patients and relatives were an ongoing active part of the development until the final product was agreed upon.

Results

In 2021, nine film cuts of 3-5 minutes premiered in an auditorium at the cinema Dagmar located in Copenhagen. Each film has a specific focus and outline complicated situations experienced from the time of being diagnosed to the establishment of having a new but changed everyday life: 1) A happy family, 2) The operation, 3) The message, 4) A new daily life, 5) Back to work, 6) Freja’s party, 7) Casework, 8) The family support group and 9) Talk about it.

Conclusion

The 9 short film cuts can be watched at The ‘Hjernetumorforeningens’ website, on YouTube, LinkedIn and on Facebook. The film cuts have gained international attention and can therefore be seen with English subtitles for international dissemination.
P09.02.B Engaging Parents in Neuropsychological Rehabilitation for Childhood Cancer Survivors: The I’M aware: Patients And Carers Together (ImPACT) Program
Authors: Wu L M , Henriksen L T , Buskbjerg C R , Lassen-Ramshad Y , Zachariae R , Evald L , Hasle H , Amidi A
Published: September 2022

Background

Cognitive impairment is the most common late effect in childhood cancer survivors (CCS). A third are likely to experience impairments in skills such as attention, processing speed, memory, and executive function. Neurobehavioral impairments are also observed in CCS (e.g., apathy) and may underlie cognitive impairment. Although these problems can impair quality of life, scholastic achievement and long-term socioeconomic outcomes, there is a dearth of research on interventions to treat cognitive impairment in CCS. Cognitive rehabilitation studies in CCS have focused primarily on computerized cognitive training and meta-cognitive strategy training, but with limited effects and poor study designs. They have lacked four key elements: 1) transfer of learning to real-world functional outcomes; 2) awareness training to enhance compensatory strategy use, particularly since CCS may experience neurological unawareness preventing the optimal application of strategies; 3) inclusion of parents that could enhance transfer of learning to the home; and 4) recognition of the inter-related nature of cognitive, neurobehavioral, and psychological functions. A novel, context-sensitive neuropsychological rehabilitation program has been developed for CSS to address these missing elements - the “I’M aware: Patients and Carers Together (ImPACT)” program - based on an evidenced-based approach pioneered at NYU Hospital in the US. In this protocol, the feasibility, acceptability, and preliminary efficacy of the ImPACT program (8 x 1 hour sessions) will be evaluated.

Material and Methods

Phase One will consist of semi-structured interviews with 5 to 10 CCS and their parents in order to review and refine procedures for the ImPACT program. In Phase Two, twenty participants will be randomized to a wait-list control (WLC) or intervention group. WLCs will not receive the intervention immediately, but will serve as the control comparison group. Measures of self- and parent-reported cognition, neurobehavioral function, and global function will be administered pre- and post- intervention, and 9 weeks later (or equivalent time points for WLCs).

Results

Results of this project will provide vital data in support of a future large-scale trial.

Conclusion

Findings could have implications for the management of cognitive and neurobehavioral impairment in CCS that could transform the nature of services provided to CCS. The program can be delivered remotely or in-person and harnesses existing supports in the child’s life to promote real-world functioning.
P09.03.A Associations of levetiracetam use with the safety and tolerability of chemoradiotherapy for patients with newly diagnosed glioblastoma
Authors: Seystahl K , Oppong F B , Le Rhun E , Hertler C , Stupp R , Nabors B , Chinot O , Preusser M , Gorlia T , Weller M
Published: September 2022

Background

Levetiracetam (LEV) is one of the most frequently used antiepileptic drugs (AED) for brain tumor patients with seizures. We hypothesized that toxicity of LEV and temozolomide-based chemoradiotherapy may overlap.

Patients and Methods

In a retrospective analysis of individual patient data using a pooled cohort of patients with newly diagnosed glioblastoma included in clinical trials prior to chemoradiotherapy (CENTRIC, CORE, AVAglio) or prior to maintenance therapy (ACT-IV), we tested associations of hematologic toxicity, nausea or emesis, fatigue, and psychiatric adverse events during concomitant and maintenance treatment with the use of LEV alone or with other AED versus other AED alone or in combination versus no AED use at the start of chemoradiotherapy and of maintenance treatment.

Results

Of 1681 and 2020 patients who started concomitant chemoradiotherapy and maintenance temozolomide, respectively, 473 and 714 patients (28.1% and 35.3%) were treated with a LEV-containing regimen, 538 and 475 patients (32.0% and 23.5%) with other AED, and 670 and 831 patients (39.9% and 41.1%) had no AED. LEV was associated with higher risk of psychiatric adverse events during concomitant treatment in univariable and multivariable analyses (RR 1.86 and 1.88, p<0.001) while there were no associations with hematologic toxicity, nausea or emesis, or fatigue. LEV was associated with reduced risk of nausea or emesis during maintenance treatment in multivariable analysis (HR=0.80, p=0.017) while there were no associations with hematologic toxicity, fatigue, or psychiatric adverse events.

Conclusion

Any association of psychiatric adverse events with LEV did not persist beyond the concomitant treatment phase. Antiemetic properties of LEV may be beneficial during the maintenance temozolomide.
P09.06.B The effectiveness of antiepileptic drug tripletherapy in refractory epileptic glioma patients: a multicenter observational cohort study
Authors: van der Meer P B , Dirven L , Fiocco M , Vos M J , Kouwenhoven M C M , van den Bent M J , Taphoorn M J B , Koekkoek J A F
Published: September 2022

Background

About 10% of the glioma patients need antiepileptic drug (AED) tripletherapy due to refractory epilepsy. This study aimed to determine whether levetiracetam combined with valproic acid and clobazam (LEV+VPA+CLB), a commonly prescribed tripletherapy, has favourable effectiveness compared to other tripletherapy combinations in glioma patients.

Material and Methods

In this multicenter retrospective observational cohort study, the primary outcome was the cumulative incidence of treatment failure for any reason, from initiation of AED tripletherapy. Secondary outcomes included cumulative incidences of: 1) treatment failure due to uncontrolled seizures; 2) treatment failure due to adverse effects; and 3) recurrent seizure. Maximum duration of follow-up was 36 months.

Results

Ninety patients, from an initial cohort of n=1435 patients, received AED tripletherapy after second-line AED duotherapy treatment failure due to uncontrolled seizures. LEV+VPA+CLB was prescribed to 43 (48%) and other AED tripletherapy to 47 (52%) patients. The cumulative incidence of treatment failure for any reason of LEV+VPA+CLB did not significantly differ from other tripletherapy combinations (12 months: 47% [95%CI, 31-62%] versus 42% [95%CI, 27-56%], p=0.892). No statistical significant differences for treatment failure due to uncontrolled seizures (12 months: 12% [95%CI, 4-25%] versus 18% [95%CI, 8-30%], p=0.445), due to adverse effects (12 months: 22% [95%CI, 11-36%] versus 15% [95%CI, 7-27%], p=0.446), or recurrent seizure (1 month: 65% [95%CI, 48-78%] versus 63% [95%CI, 47-75%], p=0.911) were found.

Conclusion

LEV+VPA+CLB has not shown favourable efficacy or tolerability compared to other AED tripletherapy combinations in glioma patients. Therefore, we cannot specifically recommend CLB as tripletherapy add-on AED to LEV+VPA.
P09.07.A Signs and Symptoms in the last days of life of the glioblastoma patients - A Descriptive and Retrospective Study
Authors: Ferreira M , Vasconcelos L , Lacerda M , Costa J
Published: September 2022

Background

Despite scientific advances to develop techniques for early diagnosis and treatment of glioblastoma, the disease continues to have a remarkably poor prognosis. Studies have shown that in patients with glioblastoma, not only the presence of end-of-life signs and symptoms but also their frequency differs from those in the rest of cancer patients. The complexity of glioblastoma symptoms makes the integration of a palliative care philosophy essential.

Objective

To identify the main signs and symptoms that occur in patients with glioblastoma in their last seven days of life in a neuro-oncology ward.

Material and Methods

This was a descriptive and retrospective study regarding the signs and symptoms in the last seven days of life of patients with glioblastoma who were admitted to the neurology ward of a national oncology reference center. The patients were admitted after neuro-oncologist consultations between 2019 and 2020. The data collection instrument used was based on the document “The Last Hours of Living”. Confidentiality of the participants was guaranteed, and authorization was obtained from the institution’s ethics committee.

Results

The sample had 17 patients (13 men and 4 women), with an average hospital stay of 17.3 days. The men and women had average ages of 61.8 and 63 years, respectively. Clinical and nursing records revealed that the most prevalent signs and symptoms were “decreased level of consciousness” (94%); “rare and unexpected events” (94%); “respiratory dysfunction” (88%); and “loss of ability to swallow” (76%). In the last seven days of life, the approximate times of presentation for these signs and symptoms were: decreased level of consciousness, 6 days; rare and unexpected events, 2 days; respiratory dysfunction, 3 days; and loss of ability to swallow, 6 days. Support was requested from the in-hospital palliative care support team for 76.4% of patients.

Conclusion

The identification of reduced level of consciousness and alterations in swallowing as the most prevalent symptoms is consistent with the findings of previous studies. The prevalence of rare and unexpected events (such as fever, myoclonus and seizures) aligns with previous studies that indicated a difference in the signs and symptoms presented by patients with glioblastoma in the last days of life and patients with other oncological diseases. In this sense, the instrument used for data collection may not be the most appropriate for the population with primary central nervous system tumors. The decreased level of consciousness and impaired communication can complicate the assessment of signs and symptoms, such as pain, nausea, anxiety and depression. It is essential to conduct research studies with more representative samples. The recording of the signs and symptoms presented, as well as interventions and the evaluation of their effectiveness, are essential to ensure the quality of end-of-life.
P10.02.B The fragile balance between proliferation and neoplasia in neural stem cells: role of DIAPH3 in glioblastoma
Authors: Chehade G , Ruiz-Reig N , Raftopoulos C , Tajeddine N , Tissir F
Published: September 2022

Background

DIAPH3 belongs to the formin family of proteins that are master regulators of the cell cytoskeleton. DIAPH3 plays a key role in cytokinesis as well karyokinesis. Hence, in the mouse brain, DIAPH3-deficient neural stem cells exhibit abnormal cell division leading to aneuploidy and apoptosis of cells progeny. Given the importance of aneuploidy in cancer development and progression, we asked whether DIAPH3 loss could be involved in the genesis and the aggressiveness of brain tumors.

Material and Methods

We co-inactivated Diaph3 and Trp53 genes in the mouse cerebral cortex using the Emx1-Cre mouse line (Diaph3/Trp53 dcKO). As a control, we inactivated only Trp53 gene using the same Cre mouse line (Trp53 cKO). Mouse survival was analyzed, and tumors were collected for histological analysis as well as molecular characterization. Additionally, we measured DIAPH3 expression in human glioblastoma samples and studied its impact on patient overall survival.

Results

Diaph3/Trp53 dcKO develop brain tumors earlier compared to Trp53 cKO resulting in a decrease of mouse survival. Hematoxylin-eosin staining, as well as GFAP, OLIG2 and KI67 immunostainings strongly suggest that the tumors are glioblastomas. Quantitative reverse transcription PCR performed on biopsies from 73 patients uncover variable DIAPH3 expression levels in glioblastoma samples. Interestingly, multivariate Cox proportional-hazards analysis shows that low DIAPH3 expression predicts a worse overall survival, independently from other known prognosis factors.

Conclusion

On one hand, our study shows that DIAPH3 loss favors glioblastoma development in mouse, strongly suggesting a tumor-suppressor role for DIAPH3. On the other hand, the clinical study shows that DIAPH3 low expression has a negative impact on patient overall survival indicating that DIAPH3 loss may favor resistance to chemotherapy and/or radiotherapy.
P10.06.A Expression of GDNF and its receptors in glioblastoma
Authors: Ewald J D , Knudsen A M , Kristensen B W
Published: September 2022

Background

Neurotrophic factors are important for brain development, but their role in glioblastoma, the most aggressive primary brain cancer, is not fully understood. We focused on glial cell line-derived neurotrophic factor (GDNF), which has been shown to play a role in growth and migration in glioblastoma. The aim of this study was to investigate the expression of GDNF and its primary receptor GFRA1 as well as GDNF family receptors GFRA2-4 and RET.

Material and Methods

Tissue sections from 10 glioblastoma patients were stained with antibodies against GDNF (frozen tissue) and each of the receptors (formalin-fixed paraffin-embedded tissue). Moreover, the sections were stained with an RNA Scope assay for GDNF mRNA. The area fractions of positive receptor staining were quantified with a software-based classifier. Double immunofluorescence stainings were performed for GFRA1 and the astrocytic tumor cell marker GFAP, the microglia and macrophage marker IBA1, and the tumor stem cell markers OLIG2 and SOX2, respectively, for assessment of co-expression.

Results

GDNF staining in a varying number of cells was seen in 5 of the 10 samples. GDNF mRNA stainings were observed in a varying number of cells in all stained tissues, including in hypoxic areas and in microvascular proliferations. Mean area fractions of the receptors were 31 % (range 7-56) for GFRA1, 3 % (0,2-7) for GFRA2, 0,4 % (0,01-2) for GFRA3, 0,1 % (0,02-0,3) for GFRA4 and 1 % (0,1-2) for RET. Co-expression of GFRA1 and GFAP was widely present. Co-expression of GFRA1 and either OLIG2 or SOX2 was partially present. Co-expression of GFRA1 and IBA1 was limited.

Conclusion

Our results suggest that GDNF and GFRA1 are widely expressed in glioblastoma tissue, but GFRA2-4 and RET are also expressed and may play a role in glioblastoma tumor biology. Future experimental studies are needed to clarify the functional role of GDNF and its receptors in glioblastoma biology.
P10.07.B Differential YAP activity in human glioblastoma tumorspheres
Authors: Kim E , Oh Y , Kang S
Published: September 2022

Background

Although glioblastoma (GBM) is the most common primary brain tumor, the best available treatment options are still associated with poor prognosis. Recently, the Hippo/YAP signaling pathway has been emerged as an important driver of GBM. Nevertheless, extensive studies have not yet been focused on the importance of phosphorylation event in regulating endogenous YAP activity in GBM. Here, we sought to elucidate that the modulation and stabilization of YAP/TAZ in Hippo pathway promotes GBM progression.

Material and Methods

Expression patterns of YAP1, TAZ, CTGF, TEAD4 and LATS1 mRNA were confirmed in human normal and GBM tissue, and patient survival according to YAP1 expression was confirmed in TCGA and Severance cohort. The YAP1 expression patterns in several TSs were classified into SOH and AH according to transcriptome analysis and western blot. For each classified TSs, the characteristics of SOH and AH were comparatively analyzed through immunoprecipitation, nucleus fraction, TEAD4 luciferase assay, and confocal analysis. The effect of YAP1 knockdown/out and overexpression in SOH and AH TSs were measured by WST/ATP analysis, 3D invasion and neurosphere formation, and western blot. In addition, the drug reactivity of SOH and AH TSs was examined by treatment with the YAP1 inhibitors Peptide17 and verteporfin, and the effects of Peptide17, verteporfin and YAP1-shRNA were examined in in-vivo xenograft model.

Results

YAP1 mRNA levels including TAZ, CTGF, TEAD4 and LATS1 were higher in GBM than in normal tissues. Through transcriptome analysis and western blot, TS can be divided into SOH and AH groups according to the expression level of CTGF, but not YAP1 level. In the case of SOH TS, the binding of TEAD4 was strong in immunoprecipitation, the ratio of YAP1 was higher in the nucleus in the nucleus fractionation and confocal analysis, and the luciferase activity of TEAD4 was high in the TEAD4 luciferase assay. On the other hand, in the case of TS of AH, the opposite trend was observed. When YAP1 was knocked down or out, the proliferation, invasion, and stemness of TS tended to be decrease more in SOH TS than in AH TS, and SOH TS showed more sensitive drug response even in Peptide17 and Verteporfin treatment. In the case of YAP1 overexpression, the opposite trend was observed in SOH TS. In the in-vivo xenograft model, injection of Peptide17, verteporfin, and YAP1-shRNA showed a tendency to increase the survival rate of mice.

Conclusion

The association of YAP has been described in several cancers, but there has been no study comparing GBM TS by YAP expression and classifying it at the functional level. Now, for the first time, we show that the characteristics of GBM TS can be predicted according to the expression value of YAP, respectively, and YAP is expected to be applied as an important prognostic factor for GBM treatment in the future.
P10.08.A Lipogenesis inhibition by fatostatin shows effectiveness in glioblastoma models highly expressing fatty acid synthase (FASN)
Authors: Kastler L , Neumayer K , Laaber M , Roschger C , Lenz S , Lötsch-Gojo D , Berger W , Gruber A , Spiegl-Kreinecker S
Published: September 2022

Background

Glioblastoma (GBM) is the most common and most aggressive primary brain tumour in adults. Despite intensive treatment, the medium overall survival of GBM patients remains below 20 months after diagnosis. Therefore, new tumour specific targets allowing successful therapeutic treatment are needed to distinctly enhance GBM patient overall survival. Reprogramming of lipid metabolism leading to highly upregulated anabolic pathways, like phosphatidylinositol-3-kinase (PI3K)/AKT pathway and subsequently lipogenesis, is a hallmark of cancer cells including glioblastomas to meet increased energy demands for proliferation and cell growth. Accordingly, lipogenesis inhibition might be a promising strategy to impede GBM cell growth. Therefore, it was aim of our study to analyse the effect of the lipogenesis inhibitor fatostatin on downstream targets e.g. fatty acid synthase (FASN) and its impact on cell growth in GBM cells in vitro.

Material and Methods

In a cohort of 52 GBM-derived cell lines, FASN mRNA was investigated by qRT-PCR. The impact of fatostatin on cell viability (IC50) was analysed in selected cell lines using cell viability assays (Cell Titer Glo®). To elucidate whether fatostatin exposure affects cell migration, wound healing assays were performed. The effect of fatostatin on cell cycle and apoptosis was analysed by flow cytometry. Additionally, expression of FASN and level of phosphorylated AKT, the active form of AKT serine/threonine kinase, were examined before and after fatostatin treatment with qRT-PCR (mRNA) and Western blot (protein), respectively.

Results

FASN mRNA was found to be expressed at varying levels in our GBM-derived cell line cohort (n=52). Compared to normal astrocytes, FASN mRNA expression was significantly upregulated in GBM cells (p=0.0127). Furthermore, high FASN mRNA expression was significantly associated with higher sensitivity against fatostatin (p=0.0352). Regarding corresponding recurrences, the response to the inhibitor was shown to be persistent. Accordingly, the effectiveness of fatostatin treatment resulted in substantial downregulation of AKT phosphorylation and consequently in the induction of apoptosis and G2/M arrest of the cell cycle. In addition, a significant inhibition of the migration potential by fatostatin treatment was observed.

Conclusion

In summary, FASN expression represents a promising biomarker and therapy target within the lipid metabolism indicated by a significant sensitivity to fatostatin in FASN overexpressing GBM cell lines. Consequently, lipogenesis inhibition by fatostatin might be a promising novel therapeutic option in glioblastoma.
P10.09.B Retroelement expression in glioma tumors exhibits subtype specific patterns
Authors: Rodriguez Almaraz J , Guerra G , Wendt G , Chang S , Francis S S
Published: September 2022

Background

There are ~3 million transposable elements in the human genome constituting about 42% of all basepairs. Retroelements (REs) are ~90% of the transposable elements present in the human genome. Active REs are considered highly mutagenic and have been implicated in multiple steps of cancer development and progression, as well as in neurologic diseases. RE activity has functional effects on the genome, including the maintenance of centromere and telomere integrity, and deleterious gene expression. Previous studies have shown that certain families of RE (HERVK, L1, Alu) are expressed in gliomas, however, their specific role as arbitrators of oncogenesis or promoters of the innate anti-tumor immune response remains uncertain. Moreover, it has been shown that a soluble form of PD-L1 (sPD-L1) that blocks its inhibitory activity is produced by exaptation of an intronic endogenous retroelement (LINE-2A) in the gene encoding PD-L1, highlighting the importance of REs as potential therapeutic targets. In this analysis we aim to identify the unique patterns of RE expression across major subtypes of glioma.

Material and Methods

We conducted a differential expression analysis of 49 RE families using RNA-seq data measured in glioma tumors from The Cancer Genome Atlas (TCGA). RE counts were produced using the software REDiscoverTE. Pairwise comparisons between glioma subtypes (defined by WHO2021) were done using in 625 tumor samples adjusting for age, sex and race.

Results

10 of the 49 considered RE families exhibited significantly different (false discovery rate, FDR, <0.05) expression in at least one glioma subtype. Alu(Fold change, FC=1.5), RNA(FC=11.3), PiggyBac(FC=1.6), rRNA(FC=5.23) and Dong-R4(FC=1.8) were overexpressed in IDH-wildtype glioblastoma while Gypsy(FC=0.4) and CRP1(FC=0.26) were decreased in expression. scRNA (FC=2.7) were overexpressed in IDH-mutant oligodendroglioma compared to glioblastoma while Dong-R4 (FC = 0.53) showed decreased expression. LTR (FC=2.02) and tRNA-Deu (FC=1.46), showed increased expression in IDH-wildtype diffuse astrocytomas compared to IDH-mutant, 1p/19q-codeleted oligodendrogliomas while Gypsy (FC =0.41) showed decreased expression.

Conclusion

We have shown that expression of certain RE families within gliomas have subtype-specific patterns. While it is well established that RE expression is dysregulated in cancer, our analysis is the first at exploring a wide range of retroelements in the context of glioma by subtype. Given the important role of REs in transcriptional control, genomic instability, chromosomal rearrangements, and oncogenic activation, the identification of individual families and specific REs in glioma holds an intrinsic value to potential biomarkers and immunotherapy targets.
P10.11.B Re-sensitizing cancer cells to Tumor Treating Fields (TTFields) through PI3K/AKT/mTOR pathway inhibition
Authors: Klein-Goldberg A , Voloshin T , Zemer-Tov E , Paz R , Koren L , Wainer-Katsir K , Wolfbauer C , Haber A , Giladi M , Weinberg U , Palti Y
Published: September 2022

Background

Tumor Treating Fields (TTFields) are alternating electric fields disrupting cellular processes critical for cancer cell replication and tumor progression. TTFields therapy is approved for treatment of newly diagnosed glioblastoma (GBM), recurrent GBM, and unresectable malignant pleural mesothelioma, and is currently under clinical investigation for treating other solid tumors, such as ovarian cancer, non-small cell lung carcinoma (NSCLC), and hepatocellular carcinoma (HCC). The research herein aimed to identify potential treatment escape mechanisms and explore the possibility of targeted inhibition of these pathways for re-sensitizing the cells to TTFields.

Material and Methods

GBM U-87 MG, ovarian A2780, and NSCLC H1299 cells were treated with TTFields (1.7 V/cm RMS, 200 or 150 kHz) for continuous long-term application (7 or 13 days, specific conditions depending on the cell line). Changes in signaling pathways in these cells relative to cells exposed to short term TTFields application (3 or 7 days, depending on cell line) were examined by Luminex multiplex assay. Specific pathway markers were examined by immunohistochemistry of tumor sections from sham or TTFields-treated rats bearing N1S1 HCC tumors. TTFields were then co-applied with relevant pathway inhibitors, followed by cell count measurements and western blot examinations for specific pathway markers. The concomitant application of TTFields with a selected inhibitor was tested in mice inoculated orthotopically with MOSE-L firefly luciferase (FFL) ovarian cancer cells. Tumor volume was measured at study end by luciferin signal detection using the In Vivo Imaging System (IVIS).

Results

Cancer cells exposed to long-term application of TTFields displayed decreased sensitivity to TTFields. The PI3K/AKT/mTOR pathway was activated in these cells, with significant increases in AKT and RPS6 phosphorylation levels also observed in HCC tumors from rats treated with TTFields. PI3K inhibitors re-sensitized the cells to TTFields cytotoxicity, with associated down regulation of AKT phosphorylation. Application of TTFields concomitant with the PI3K inhibitor alpelisib resulted in enhanced efficacy in the ovarian cancer mouse model.

Conclusions

The current research demonstrated that PI3K/AKT/mTOR signaling pathway activation was involved in the response to long-term application of TTFields, with increased phosphorylation of key proteins observed both in vitro and in vivo. Concomitant treatment with PI3K inhibitors and TTFields re-sensitized the cells to TTFields, as it alleviated TTFields-induced AKT activation.
P10.16.A Repurposing thioridazine as a novel treatment of melanoma brain metastasis
Authors: Moen Taule E , Wikerholmen T , Rigg E , Sarnow K , Wang J , Thorsen F
Published: September 2022

Background

Melanoma shows increasing incidence and has the highest propensity to metastasize to the brain of all primary malignant tumors. Untreated, the median survival time is 2-3 months after diagnosis, while aggressive treatment extends survival to only 4-12 months. Even though targeted therapies and immunotherapy has changed the therapeutic landscape, many of these patient relapse. In this respect, drug repurposing, using old drugs for new purposes, has had several successes for other diseases. The anti-psychotic drug thioridazine (THD) is a dopamine receptor 2 antagonist, with properties to cross an intact blood brain barrier, has shown promising effect on melanoma cells that has escaped prior treatment and on cancer stem cells. However, the therapeutic effects on melanoma brain metastasis (MBM) have not been previously investigated.

Material and Methods

All experiments were done on MBM cell lines (H1, H2, H3 and H10), which were developed in our lab from patient biopsies obtained after surgery. A monolayer viability assay (WST-1) was used to determine IC50 doses. Migration and proliferation after treatment was studied by IncuCyte live cell imaging. Colony forming properties and the ability of cells to grow in an anchorage free environment after THD treatment was investigated using a 2D clonogenic assay and a 3D soft agar model. Potential drug effects were also studied on normal human astrocytes (NHA) and on fetal rat brain organoids (FRBO), using a Live/Dead viability kit and confocal imaging. Apoptosis and necrosis was studied by Annexin V/propidium iodide (PI) double staining followed by flow cytometric analysis after 72 hours of treatment.

Results

THD shows a strong cytotoxic effect on MBM cell lines. Cell viability studies showed that THD exerted a dose-dependent inhibition of tumor cell viability (IC50 doses 9-12 µM), while NHA tolerated similar drug concentrations. Tumor cell migration and proliferation were almost completely inhibited with concentrations below the IC50 doses. An inhibition of clonogenic formation and 3D growth was also observed. Bright field microscopy showed changes in MBM morphology in monolayers, but FRBOs were intact after drug treatment. Further, apoptosis in MBM cells was induced after treatment with THD.

Conclusion

Our preliminary data shows that THD has a strong selective cytotoxicity in MBM cells with minimal effects on NHA and FBRO.
P10.18.A Replication of previous GWAS identifies TERT and near EGFR SNVs as risk factors in EPIC glioma patients: a nested case-control study
Authors: Wu W , Dahlin A , Andersson U , Wibom C , Jonsson S , Melin B
Published: September 2022

Background

Gliomas, the most common malignant primary brain tumors in adults, typically have a poor prognosis irrespective of medical care. Previous large genome-wide association studies (GWAS) have identified 27 single-nucleotide variants (SNVs) that are significantly associated with glioma. However, most of the GWAS are conducted by case-control study designs, it is therefore prone to bias when rapidly lethal cases don’t have chance to be included in the study. This study aims to replicate the previous GWAS findings using prospective study design.

Material and Methods

We conducted a nested case-control study within the European Prospective Investigation into Cancer (EPIC) cohort from 7 European countries. GSA-MD Infinium global screening array was used for genotyping. Some subjects were genotyped by other platforms previously. In total, 468 glioma patients and 481 controls were included. The genotypes of 27 SNVs were extracted and for ungenotyped SNVs, datasets were imputed using SHAPEIT v4.1.3 and IMPUTE5 v1.1.5 based on the Haplotype Reference Consortium (Release 1.1) reference panel. Conditional logistic regression model was used to investigate the additive effect of SNVs on the risk of glioma.

Results

21 SNVs showed a consistent direction of effect with previous studies, whereas 6 SNVs did not (ORs between 0.72-0.99 and not significant). After adjusting for multiple testing, two SNVs, rs10069690 (TERT), and rs75061358 (near EGFR) were significantly associated with glioma risk. We observed that prominent OR (2.23, 95%CI=1.49-3.33) of rs75061358 in our study compared to the result from previous GWAS, which implied rs75061358 might be not only a risk factor but also affect survival. Different risk direction was observed for rs77633900 in ETFA gene (OR=0.72, 95%CI=0.51-1.01).

Conclusion

Our findings further confirmed the genetic role on the etiology of glioma in the European population. The potential biases from the previous GWAS are required to be elucidated.
P10.20.A Mechanisms of synergistic glioma growth suppression by radiotherapy and MET inhibition
Authors: Silginer M , Papa E , Szabo E , Vasella F , Pruschy M , Stroh C , Roth P , Weiss T , Weller M
Published: September 2022

Background

Glioblastoma remains to be one of the most lethal solid cancers. Despite multi-modal therapy including surgery as safely feasible, radiotherapy and chemotherapy with the alkylating agent temozolomide, the median survival of affected patients is still limited to approximately one year on a population level. Thus, novel therapies are urgently needed. There is increasing interest in the role of the HGF/MET pathway in the response of glioblastoma to radiotherapy since MET may be involved in radioresistance via proinvasive and DNA damage response pathways.

Material and Methods

Here we assessed the role of the MET pathway in the response to radiotherapy in vitro and in vivo in syngeneic mouse glioma models and explored potential modes of action responsible for the synergistic effects of MET pathway inhibition and irradiation on tumor growth in vivo.

Results

Murine glioma cells express HGF and MET and show increased MET phosphorylation upon exposure to exogenous HGF. In vitro, glioma cell viability and proliferation are not affected by pharmacological MET inhibition using tepotinib or genetic MET inhibition using CRISPR/Cas9-engineered Met gene knockout and sensitization to irradiation by MET inhibition is not seen. In vivo, the combination of MET inhibition with focal radiotherapy mediates prolonged survival of syngeneic orthotopic glioma-bearing mice compared with either treatment alone. Complementary studies demonstrate that synergy is lost when gliomas are established and treated in immunodeficient mice, but also if MET gene expression is disrupted in the tumor of wildtype mice. Combination therapy suppresses a set of pro-inflammatory mediators that are upregulated by radiotherapy alone and which are positively regulated by transforming growth factor (TGF)-β. In line with this data, ex vivo analysis of mouse brains reveal increased TGF-β pathway activity upon irradiation alone that is counteracted by concomitant MET inhibition.

Conclusion

In summary, we demonstrate synergistic suppression of syngeneic glioma growth by irradiation and MET inhibition that requires MET expression in the tumor as well as an intact immune system. Clinical evaluation of this combined treatment approach in newly diagnosed glioblastoma is warranted.