bluuebunny/crossref_metadata_2025_split huggingface dataset:
OS08.4.A Analysis of melanoma brain metastasis immune microenvironment through multiplex gene expression profiling
Authors: Ricci A A , Collemi G , Orlando G , Tampieri C , Senetta R , Pellerino A , Bruno F , Melcarne A , Garbossa D , Rudà R , Soffietti R , Ribero S , Quaglino P , Cassoni P , Bertero L 
Published: September 2022

Abstract

Background
Novel immunotherapies based on targeting of specific immune checkpoints enabled a significant improvement of melanoma outcome, but melanoma brain metastases (BM) remain an unmet oncological need with an overall 2-year survival rate lower than 10%. Tumour immune microenvironment has been demonstrated to play a key role in BM establishment and development, but data regarding the specific milieu of melanoma BM is limited.

Material and Methods
Gene expression profiles of 55 samples of primary melanoma and BM were evaluated using the nCounter PanCancer IO 360 Panel (NanoString Technologies) targeting 770 mRNA involved in tumor immune microenvironment modulation. The case series consisted of 10 primary melanomas and their 10 matched BM, 25 unmatched BM, and 10 locally advanced control melanomas without evidence of BM after &gt;5 year follow up.

Results
Among BM samples, most patients (25/45) were males and median age at BM diagnosis was 61,2 years with a median time to BM development of 2,1 years. Median OS from BM diagnosis was 1,3 years. Several genes resulted significantly downregulated in BM compared to primary melanomas, including SERPINB5 (p&lt;0.001), ARG1 (p=0.0067), S100A8 (p&lt;0.001), S100A9 (p&lt;0.001), S100A12 (p=0.0037), IL1RN (p=0.0012), CCL21 (p=0.0012), CCL22 (p=0.0012) and CCL13 (p=0.037) and SELE (p=0.026); conversely, C7 was upregulated (p&lt;0.001). Downregulated signatures in BM involved those associated with multiple immune cell populations, including neutrophils, dendritic cells, mast cells and Treg, as well as inflammatory chemokines, the CTLA4 immune checkpoint and ARG1 enzyme function; conversely, MAGEs-related signature was upregulated. Comparison between primary melanomas which developed BM and those which did not showed a significant overexpression of RRM2 (p=0.0247) and TNFRSF1A (p=0.032) genes in the latter group and an upregulation of the PD-1 pathway. Analysis according to tumour mutational status showed an upregulation of signatures associated with inflammatory chemokines, dendritic and myeloid cells and neutrophils. No differences were observed according to time to BM development and survival from BM diagnosis.

Conclusion
Our findings show that melanoma BM harbor distinct immunosuppressive mechanisms compared to primary tumors: this data elicits the importance of investigating the heterogeneity of BM microenvironment. Genes and pathways selectively overexpressed or downregulated in melanoma BM should be validated to be possibly considered as novel therapeutic targets.
OS08.5.A Adenovirus-mediated delivery of the MHC-II Transactivator CIITA gene induces tumor cell killing in immunocompetent glioblastoma organoids
Authors: Salvato I , Klein E , Forlani G , Poli A , Oudin A , Baus V , Golebiewska A , Accolla R , Niclou S P , Marchini A 
Published: September 2022

Abstract

Background
Although immunotherapies represent an encouraging approach against cancer, to date none translated to the clinical benefit in Glioblastoma (GBM). One aspect contributing to this failure is the highly immunosuppressive GBM microenvironment. Our approach to overcome immunosuppression is to increase anti-tumor immune responses via adenovirus (AdV)-mediated delivery of the MHC-II Transactivator (CIITA) gene. CIITA-induced MHC-II expression is anticipated to convert GBM cells into surrogate antigen presenting cells able to prime T helper cells, therefore promoting CD4+ and CD8+ mediated immunity.

Material and Methods
We generated AdVs containing wild type CIITA (Ad-CIITA) using a replication-defective serotype5 adenoviral backbone. AdVs containing a mutated, non-functional version of CIITA (Ad-CIITA mutant) and an empty CMV promoter (Ad-null) were used as controls. AdV-mediated MHC-II expression was monitored at mRNA, protein and cell surface level. For the functional assessment of anti-tumor immune responses, we developed an advanced human GBM organoid model system consisting of tumor organoids co-cultured with either human peripheral blood mononuclear cells (PBMCs) or isolated CD3+ T cells. T cell mediated tumor cell killing was monitored over time via live cell imaging and flow cytometry.

Results
We successfully constructed and produced a CIITA-armed AdV that induces MHC-II expression in infected GBM cells, indicating the efficient expression of transcriptionally active CIITA for at least six days post infection. In immunocompetent human GBM organoids, Ad-CIITA infection of tumor cells led to prominent organoid disruption and tumor cell death, an effect that was not observed in the absence of PBMCs or CD3+ T cells. Tumor organoids infected with Ad-CIITA mutant remained intact, demonstrating the implication of cell surface MHC-II molecules in the observed phenotype.

Conclusion
Our results demonstrate that AdV-mediated delivery of CIITA is a promising strategy to increase T cell mediated immunity against glioblastoma.
OS08.7.A Lomustine and the immunocytokine L19TNF are a promising treatment combination for recurrent glioblastoma
Authors: Look T , Puca E , Stucchi R , Luca R , Roth P , Neri D , Weller M , Hemmerle T , Weiss T 
Published: September 2022

Abstract

Background
Treatment options for recurrent glioblastoma are limited and with the possible exception of regorafenib, no agent has demonstrated superior activity to lomustine. Therefore, there is an urgent need for more effective treatment strategies for recurrent glioblastoma. Here, we investigated different treatment combinations based on the tumor-stroma targeting antibody-cytokine fusion protein L19TNF in preclinical glioma models and translated the most effective treatment combination to patients with recurrent glioblastoma.

Material and Methods
Orthotopic immunocompetent mouse glioma models were used to study the anti-glioma activity of L19TNF in combination with anti-PD1, bevacizumab or lomustine. Tumor growth was monitored by MRI. Flow cytometry and microscopy were used to characterize tumor-infiltrating-immune cells. MHC immunoaffinity purification and mass spectrometry were used to characterize the MHC immunopeptidome. Genetic mouse models were used to study immune-dependent effects. Subsequently, we translated the most efficient treatment combination to patients with recurrent glioblastoma within a phase I/II clinical trial (NCT04573192).

Results
The combination of L19TNF and lomustine demonstrated strong synergistic anti-tumor activity in two immunocompetent orthotopic glioma models and cured a majority of tumor-bearing mice. In contrast, combinations with anti-PD-1 or bevacizumab had only limited anti-glioma activity. Furthermore, compared to the monotherapies, the combination of L19TNF and lomustine led to the strongest increase in tumor-infiltrating lymphoid cells as demonstrated by flow cytometry and microsopy and to the highest number of peptides presented in the context of MHC-I. The treatment effect was abrograted in different genetic immunodeficient mouse models. The treatment combination of L19TNF and lomustine was well tolerated in the first patients treated within a phase I/II clinical trial and we observed partial tumor responses also in patients with an unmethylated MGMT promoter.

Conclusion
The combination of L19TNF and lomustine demonstrated promising anti-glioma activity and patients are currently recruited within a phase I/II clinical trial for patients with recurrent glioblastoma.
OS08.8.A Integrin-specific CAR T cells for the treatment of glioblastoma
Authors: Villars D , Hänsch L , Silginer M , Weiss T , Weller M , Roth P 
Published: September 2022

Abstract

Background
Current standard of care for glioblastoma patients has limited therapeutic efficacy and novel innovative treatment strategies are urgently needed. One such strategy is chimeric antigen receptor (CAR) T cell therapy that has shown great success in hematological malignancies. αv integrins are overexpressed in several neoplasms and have already been used as therapeutic targets for small molecule inhibitors and antibodies, which did not cause major toxicities. Herein, we propose αv integrins as an ideal target for CAR T cell therapy in glioblastoma.

Material and Methods
CAR T cells targeting specific heterodimers (αvβ3, αvβ5 or αvβ8) were generated by transducing primary human T cells from healthy donors with a lentiviral vector expressing a second-generation CAR. Activity and specificity of CAR T cells was determined by co-culture assays with different glioma cells. Efficacy of CAR T cells to control tumor growth in vivo was investigated in clinically relevant orthotopic xenograft glioma mouse models. Additionally, we generated CAR T cells from T cells from a glioblastoma patient and measured their activity against the patient’s autologous tumor cells.

Results
All newly generated integrin-targeting CAR T cells exerted strong anti-glioma activity in vitro. Long-term and repetitive killing assays as well as cytokine-release measurements demonstrated highest activity of αvβ5 and αvβ8 integrin-specific CAR T cells. Antigen specificity of these cells was confirmed, as glioma cells with a CRISPR/Cas9-mediated knockout of the target antigen were resistant to CAR T cell-mediated cytotoxicity. Intratumoral injection of αvβ5 or αvβ8 CAR T cells significantly prolonged the survival and cured a substantial fraction of glioma-bearing mice in two different xenograft models. When used in a patient-derived setting, matched CAR T cells exerted strong anti-glioma activity.

Conclusion
We show strong and integrin-specific anti-glioma activity of CAR T cells developed from healthy donor T cells and glioblastoma-patient-derived T cells in vitro. αvβ5- and αvβ8-specific CAR T cells exerted the best therapeutic activity in two different xenograft glioma models in vivo. These data support the evaluation of integrin-specific CAR T cells as a therapeutic strategy in clinical neuro-oncology.
OS09.4.A Cognitive functioning of patients with diffuse glioma during stable disease
Authors: Gorter M , Röttgering J G , Belgers V , van Lingen M R , De Witt Hamer P C , Douw L , Klein M 
Published: September 2022

Abstract

Background
Glioma patients often experience cognitive problems, which are associated with decreased functional independence and health-related quality of life. However, the prevalence and nature of cognitive impairment in these patients is relatively underreported, particularly during stable disease. In this study we determine the prevalence of cognitive deficits and explore distinct cognitive profiles of patients with diffuse glioma at least two months after tumor treatment.

Material and Methods
A total of 112 patients (mean age: 43 years) without clinical or radiological progression with a grade II-IV glioma were included in this observational cohort study. Cognitive functioning was assessed in five cognitive domains (attention, information processing speed, verbal memory, working memory, and executive functioning). Cognitive impairment was defined as a z-score of 1.5 SD below that of healthy controls. Hierarchical cluster analysis was used to examine cognitive profiles.

Results
In total, 46 patients (41%) had cognitive impairment. More than one domain was impaired in 25 of these patients (54%). The domains were affected in decreasing order of frequency: working memory in 31%, information processing speed in 22%, attention in 19%, verbal memory in 12%, and executive functioning in 11% of the patients. A clustered heatmap revealed three prominent cognitive clusters with an increasing number of cognitive domains impaired. The first cluster displayed patients with only working memory deficits, patients in the second cluster also had deficits in information processing speed, and the last cluster showed patients with impairments across all domains.

Conclusion
Cognitive impairment is highly prevalent in patients with diffuse glioma during stable disease. Working memory and information processing speed are most frequently affected. Importantly, our analyses show evidence for three subgroups in cognitively impaired glioma patients. Working memory seems an important driver in cognitive impairment given the involvement in all subgroups.
OS11.4.A Intrathecal delivery of dendritic cell vaccine eradicates tumor growth and protects against leptomeningeal disease re-inoculation in immunocompetent HER2+ and triple negative breast cancer leptomeningeal disease xenograft models
Authors: Law V , Kodumudi K , Snyder C , Czerniecki B , Forsyth P 
Published: September 2022

Abstract

Background
Leptomeningeal disease (LMD) occurs in ~5% of patients with breast cancer (BC) and has a median survival of 2-4 months. We found a loss of the anti-HER2 and anti-HER3 CD4 Th1 immune responses in BC patients. In pre-clinical and clinical trials the administration of class II HER2 peptide-pulsed dendritic cell vaccine (HER2-DCV) partially restores anti-HER2 Th1 immune responses with pathologic complete responses in HER2+ BC patients. Here, we examined the intrathecal (IT) delivery of HER2/HER3-DCV in BC-LMD immunocompetent animal models.

Material and Methods
Luciferase-labeled HER2+ TUBO BCs were injected into the cisterna magna of BALB/c mice to produce LMD. We used our Murine Ommaya (mimics an Ommaya reservoir clinically in patients) for the IT administration of DCVs into the cerebral spinal fluid (CSF).

Results
BC-LMD mice were randomized into following groups: 1) HER2-DCV IT 2) HER3-DCV IT 3) HER2/HER3-DCV IT. The median survival of untreated (control) group was 15 days. All groups given DCV IT prolonged survival (p&lt;0.001). Interestingly, HER2-/HER3-DCV IT was able to rescue disease mice (71% in HER2+ BC-LMD and 28% in triple negative breast cancer-LMD) and showed complete tumor regression. Some surviving mice were immune to subsequent tumor rechallenge. In mice CSF, we found evidence of CD4+ and CD8+ T-cells infiltration, and robust IFN-gamma and IL18 response upon DCV treatment.

Conclusion
Our preclinical data supported a clinical trial (submitted) of the IT delivery of DCV in BC patients with LMD.
OS11.5.A Utility of lumbar puncture (LP) and cerebrospinal fluid (CSF) profile in the diagnosis of neoplastic meningitis (NM): a retrospective cohort study and meta-analyses
Authors: Tuohy K J , Major K , Aregawi D , Glantz M 
Published: September 2022

Abstract

Background
CSF cytology remains the gold standard for diagnosing NM, but the sensitivity of this test is low (approximately 50% with one LP). Little is known about the predictive value of routine CSF and clinical findings for a positive cytology. We addressed these questions by analyzing 337 consecutive patients at our institution, and by conducting exhaustive evidence-based literature reviews and meta-analyses of relevant research.

Material and Methods
337 consecutive patients at a single institution were evaluated. Of these, 210 met criteria for NM (a positive CSF cytology on at least one of four pre-treatment samples) or no NM (four consecutive negative CSF cytologies). CSF white blood cell count, glucose, protein, and lactate were collected, as well as the presence of neurological symptoms. Optimal threshold values were identified using ROC curves. Sensitivity (Sn), specificity (Sp), and likelihood ratios were calculated for each variable. Two PRISMA-compliant meta-analyses were also performed: one analyzed studies reporting the cytology results of multiple LPs, and one analyzed the frequency of abnormal CSF findings as predictors of NM.

Results
All published results describe outcomes in patients with known NM. These results are, therefore, strongly distorted by spectrum bias, which artificially inflates the apparent sensitivity and specificity of those outcomes. Thus, 25 studies report the yield of positive CSF cytology on the first, second, and third LP as 73%, 87%, and 94%, but also do not describe the parameters of the procedure. Our series in patients with suspected NM (the cohort most relevant to the practicing physician) reports 74%, 85%, and 92% for immediately processed, 10 mL or greater, lumbar and ventricular specimens. Similarly, while 50 studies suggested that low CSF glucose, high CSF protein, and suggestive symptoms are associated with NM, we found very poor Sn/Sp for elevated protein (61%/61%), low glucose (35%/85%), elevated lactate (35%/91%), and symptoms consistent with NM (74%/73%). Of these values, only an elevated lactate and symptoms were even marginally helpful in predicting a positive CSF cytology. No variable predicted a negative cytology, and 22.8% of patients with a positive CSF cytology had completely normal CSF findings.

Conclusion
Generally accepted concepts about the frequency of CSF and clinical findings in patients with suspected NM are based on biased estimates in the literature. No CSF chemistry or clinical finding reliably predicts the presence of NM, and almost a quarter of patients with cytologically proven NM have completely normal findings. In patients with positive CSF cytologies, about 73% are detected after one LP, and more than 90% after 3, but again these findings represent overestimates because of pervasive spectrum bias and differences in sample acquisition/handling. Better diagnostic techniques are desperately needed.
P01.03.B A quantitative comparison of cognitive performance and patient-reported symptoms in preoperative lower-grade glioma patients from two Dutch Hospitals
Authors: Belgers V , van Buijtene W , Butterbrod E , Röttgering J G , Douw L , De Witt Hamer P C , Rutten G M , Klein M , De Baene W , Kouwenhoven M C M , Gehring K 
Published: September 2022

Abstract

Background
Protocols for assessment of (neuro)psychological outcomes in lower-grade glioma patients vary between hospitals. This potentially complicates generalization of these outcomes. We compared standardized scores on tests of two frequently impaired cognitive domains (attention and executive functioning (EF)), and two relevant patient-reported outcomes (PROs; depression and fatigue) of two neuro-oncological hospitals that use different measurement instruments.

Material and Methods
Data were used from preoperative assessments of patients with (IDH-mut) WHO grade II/III glioma tested between 2007 and 2021 at Amsterdam UMC (AMS) or at Elisabeth-Tweesteden Hospital Tilburg (ETZ). AMS patients were referred for (neuro)psychological assessment based on physician and patient preference (paper and pencil tests), whereas all ETZ patients routinely undergo screening (computerized tests). To compare scores of the different attention and EF tests we converted patients’ performances to z-scores based on normative data. For cognitive performance, we compared scores of different cognitive flexibility tests (CST vs SAT), processing speed tests (SDC vs LDMT), and Stroop tests (Stroop I and Stroop III). PROs included the CES-D vs HADS-D and the CIS-fatigue vs MVI-general fatigue (AMS vs ETZ, resp.). Differences were tested using Fisher's, χ 2, and Mann-Whitney U tests.

Results
Assessments were done median 4 weeks (AMS, n=97, range 19-0 weeks) and 1 day (ETZ, n=106; range 14-0 days) preoperatively. Age, sex, tumor location and histology were comparable between cohorts (p&gt;0.05), but the AMS cohort showed significantly more grade III tumors (36% vs 16%) and more awake surgeries (84% vs 46%). Z-scores measuring attention and EF (n=94 and n=95, AMS vs ETZ) were not significantly different (CST vs SAT, percentage with a disorder (z &lt;-1.5SD) 15% vs 13%; SDC vs LDMT 13% vs 14%; Stroop I 11% vs 18%; Stroop III 13% vs 16% at AMS and ETZ, resp.). Percentages of patients with possible depression (CES-D≥16, n=88 and HADS-D≥8, n=106) did not differ significantly between hospitals (28% vs 26%), nor did percentages of patients with severe fatigue (CIS-fatigue≥35, n=88 and MVI-general fatigue (z &lt;-1.5SD), n=38, 42% vs 24% at AMS and ETZ, resp.).

Conclusion
Standardized scores of glioma patients on cognitive domains (attention and EF) and PROs (depression and fatigue) did not differ between two centers with slightly different samples using different testing protocols. This cautiously suggests that study findings on cognitive functioning and symptoms could be generalized. For research purposes, conjoint use of pooled populations for outcome evaluation could be explored with different samples from other centers using different instruments.
P01.05.A Working memory performance is associated with functional connectivity between the right dlPFC and DMN in glioma patients
Authors: Smolders L , de Baene W , Rutten G , van der Hofstad R , Florack L 
Published: September 2022

Abstract

Background
Patients with primary brain tumors frequently suffer from cognitive impairments in multiple domains, leading to serious consequences for socio-professional functioning and quality of life. The functional-anatomical basis of these impairments is still poorly understood.
The study of correlated BOLD activity in the brain (i.e. functional connectivity) has greatly contributed to our understanding of how brain activity supports cognitive function. In particular, activity observed during the execution of specific tasks can be related to various distributed functional networks, stressing the importance of interactions between remote brain regions. Among these networks, the Default Mode Network (DMN) and the Fronto-Parietal Network (FPN) have consistently been associated with working memory performance.
Recently, using task-fMRI in glioma patients, poor performance in a working memory task was associated with less deactivation of the DMN during this task and to a lack of task-evoked changes in the DMN-FPN structure. In this study, we investigated whether these effects are reflected in the resting-state (RS) functional connectivity of the same patient group, i.e. when no task was performed during fMRI. We additionally zoomed in on the part of the FPN located in the dorsolateral Prefrontal Cortex (dlPFC), since this region is believed to be mainly responsible for DMN deactivation.

Material and Methods
Resting-state functional MRI data were acquired pre-operatively from 45 brain tumor patients (20 low- and 25 high-grade glioma patients). Results of a pre-operative in-scanner N-back working memory fMRI task were used to assess working memory performance.
Patient brains were parcellated into ROIs using both the Gordon and Yeo atlas, which have the FPN and DMN network identities readily available. The dlPFC was defined based on masks retrieved from NeuroSynth.
To measure DMN-FPN functional connectivity the average Pearson correlation between the activation time series in the regions belonging to the FPN and the DMN was calculated. Functional connectivity between the DMN and the dlPFC was calculated in a similar way.

Results
The average correlation between the resting-state fMRI activity in the right dlPFC and in the DMN was negatively associated with working memory performance for both the Gordon atlas (p &lt; 0.003) and Yeo atlas (p &lt; 0.007). No association was found for the correlation between activity in the left dlPFC and the DMN, nor for the correlation between the activity in the whole FPN and the DMN.

Conclusion
Our findings show that working memory performance of glioma patients is related to interactions between networks that can be measured with resting-state fMRI. Furthermore, the results provide further evidence that not only specific brain regions are important for cognitive performance, but that also the interactions between large-scale networks should be considered.