bluuebunny/crossref_metadata_2025_split huggingface dataset:
TMIC-07. PYK2 SIGNALING IS UPREGULATED IN RECURRENT GLIOBLASTOMA TUMORS IN C57BL/6/GL261 GLIOMA IMPLANTATION MODEL
Authors: Ortiz-Rivera Jescelica 
Published: November 2022

Abstract
Glioblastoma (GBM) is the most aggressive type of brain cancer with no successful treatment available. Microglia makes up to 30% of total GBM tumor mass. Previously we demonstrated that microglia within the tumor stimulate glioma cell growth and invasion through the proline rich tyrosine kinase 2 (Pyk2). Tumor resection is the first line treatment for GBM. Our previous studies with use of mouse glioma implantation model indicate that microglia in a site of tumor resection undergo significant modulations in cytokines expression profile. We hypothesize that microglia activated by tumor resection induces Pyk2 signaling in glioma cell and promotes malignant properties of the tumor cells that fail to be eliminated by resection. Using C57BL/6/GL261 mouse glioma implantation model, we analyzed primary implanted tumors and re-grown tumor after surgical resection. Two weeks after glioma cells implantation, tumor resection was performed, and two weeks later regrown tumors were used for western blot analysis. Total Pyk2 and phosphorylated Pyk2[Y579/50] were analyzed. The study revealed that Pyk2 phosphorylation is upregulated in re-grown tumor compared to primary implanted tumor. Treatment with PF-562271 (Pyk2/FAK inhibitor, once/daily, 50mg/kg, orally) beginning the 2ndday after tumor resection, reduced Pyk2 phosphorylation in regrown tumors compared to animals that did not receive treatment with PF-562271. In conclusion we can state that tumor resection procedure induces the upregulation of Pyk2 phosphorylation in regrown tumors. This effect is eliminated with oral administration of PF-562271.
TMIC-09. HV1 PROTON CHANNELS PROMOTE MYELOID INFILTRATION AND GLIOMA PROGRESSION
Authors: Zheng Jiaying , Wang Lingxiao , Ayasoufi Katayoun , Zhao Shunyi , Goddery Emma , Fain Cori , Johnson Aaron , Wu Long-Jun 
Published: November 2022

Abstract
Myeloid cells comprising up to 50% of total tumor mass in glioblastoma (GBM) contribute to tumor progression and immunosuppression. Restraining glioma-induced myeloid cell infiltration increases functional T cells within the tumors, improves the efficacy of checkpoint blockade, and significantly extends mouse survival. Here, we focus on an ion channel, voltage-gated proton channel Hv1, which is mainly expressed in myeloid cells and shapes the physiological functions of myeloid cells. Our bioinformatics analysis demonstrated that elevated Hv1 expression in the tumor mass correlates with poor disease prognosis in patients. Using the leading immunocompetent model of GBM in mice, we showed that Hv1 knockout mice (Hv1-/-) exhibit slower glioma progression and prolonged survival. Notably, in vivo two-photon imaging, immunofluorescence, and full-spectrum flow cytometry revealed that Hv1-/- mice have reduced monocyte/macrophage infiltration, higher frequency of MHCII+ among all infiltrating myeloid cells, and increased PD-1+CD4+ T cells in the tumor-burdened hemisphere. As a result, we combined anti-PD-1 treatment with Hv1 knockout in the middle stage of glioma, and found that approximately 30% of Hv1-/- mice were cured. Together, our results demonstrated that Hv1 regulates myeloid infiltration and could be a novel therapeutic target for the treatment of GBM.
TMIC-10. IDENTIFICATION, VALIDATION AND BIOLOGICAL CHARACTERIZATION OF NOVEL GLIOBLASTOMA TUMOUR MICROENVIRONMENT SUBTYPES: IMPLICATIONS FOR PRECISION IMMUNOTHERAPY
Authors: White Kieron , Connor Kate , Meylan Maxime , Bougoüin Antoine , Salvucci Manuela , Bielle Franck , O’Farrell Alice , Sweeney Kieron , Weng Linqian , Bergers Gabriele , Dicker Patrick , Ashley David , Lipp Eric S , Low Justin , Zhao Junfei , Wen Patrick Y , Prins Robert , Verreault Maite , Idbaih Ahmed , Prehn Jochen , Varn Frederick , Verhaak Roel , Sautès-Fridman Catherine , Fridman Wolf , Byrne Annette 
Published: November 2022

Abstract
New precision medicine therapies are urgently required for glioblastoma (GBM). However, to date, efforts to subtype patients based on molecular profiles, have failed to direct treatment strategies. We hypothesized that interrogation of the GBM tumor microenvironment (TME) and identification of novel TME-specific subtypes could inform new precision treatment strategies. To this end, a refined and validated microenvironment cell population (MCP)-counter method was applied to &gt; 800 GBM patient tumours and validated by multiplex-immunohistochemistry. The MCP-counter deconvolution method interrogates the TME composition from transcriptomic data. Using this refined method, we classified the GLIOTRAIN(www.gliotrain.eu) IDHwt GBM cohort (n=123) into 3 novel clusters characterised by differences in TME composition and subsequently validated findings in the TCGA (n=69), CGGA (n=72) and DUKE (unpublished)(n=162) cohorts. TMEHigh tumours (30%) displayed elevated immune populations, functional orientation markers, immune checkpoint genes, and upregulated immunoregulatory pathways. Moreover, tertiary lymphoid structures were a feature of TMEHigh/mesenchymal+ patients. TMEMed (46%) tumours displayed heterogeneous immune populations and upregulated neuronal signalling pathways. TMELow (24%) tumours represented an ‘immune-desert’ group, high EGFR mutation frequency and upregulated EGFR signalling pathways. Longitudinal analysis of the GLASS cohort revealed TME-subtype transitions upon recurrence, influenced by TME composition changes. Finally, assessment of three GBM immunotherapy clinical trial cohorts revealed that TMEHigh patients treated with neo-adjuvant anti-PD1 have a significantly improved survival (P=0.04). Moreover, TMEHigh patients treated with anti-PD1 and an oncolytic virus (PVSRIPO) in the adjuvant setting, showed a trend towards improved survival (P=0.15 and P=0.056 respectively). Overall, we have established a novel TME-based classification system for application in intracranial malignancies. This system may be used to better inform a precision targeting approach in the brain tumour setting. For example, we hypothesise that patients bearing TMELow tumours may be amenable to neoadjuvant anti-TIM3 + EGFR inhibitor, TMEMed to anti-angiogenic immunotherapy, and TMEHigh patients to neoadjuvant anti-PD1 + anti-CTLA4.
TMIC-17. DISCOVERING DOMINANT TUMOR CHARACTERISTICS IN CYSTIC GLIOBLASTOMA
Authors: Chen Zhihua , Miao Yifeng , Yao Shun , O’Meally Denis , Frankhouser David , Tang Biao , Li MingHui , Frankel Paul , Shahmanyan Mari , Qiu Yongming , Rockne Russell C 
Published: November 2022

Abstract

BACKGROUND
As a rare subtype of glioblastoma (GBM), the genetic features and outcomes in cystic GBM (cGBM) are largely unknown. This study aimed to evaluate the survival of cystic features, investigate genomic patterns and the immune microenvironment in patients with cGBM.

METHODS
Gd T1-weighted, T2-weighted, T2-Flair, DWI or ADC images were used to classify cGBM or noncystic GBM (non-cGBM). Clinical information from IvyGAP, EGA databases and Chinese cohort were used for survival analysis. Fresh cystic fluid and residual tumor tissue were collected to evaluate immunological components, pathological and genomic differences between cGBM and non-cGBM. Data from IvyGAP were included to determine difference of molecular feature among substructure.

RESULTS
143 cases with cGBM and non-cGBM were screened to compare survival outcomes. Cystic features were to confer a survival benefit. It was an independent prognostic factor irrespective of IDH, MGMT and therapeutic regimen. cGBM patients have a hypomethylation state compared to non-cGBM. Active immune cells such as CD4+, CD8+ T cells were present in the cystic fluid and infiltrated into tumor. 10 cytokines/chemokines were only detected in cystic fluid, 8 cytokines/chemokines increased two-fold in cystic fluid than in peripheral blood serum (PBPs). Much differential methylated position (DMPs), differentially expressed genes (DEGs), Gene Set Enrichment Analysis (GSAE), Functional Epigenetic Modules (FEM) networks and molecular feature of substructural regions were different between cGBM and non-cGBM.

CONCLUSION
These results imply that cGBM could be a new subtype and has distinctive immune microenvironment which may have potential value correlating with prognosis.
TMIC-20. SERINE AND ONE-CARBON METABOLISM ADAPTING GLIOMA CELLS TO LOW GLUTAMINE MICROENVIRONMENT
Authors: Tanaka Kazuhiro , Nagashima Hiroaki , Iwahashi Hirofumi , Hashiguchi Mitsuru , Sasayama Takashi 
Published: November 2022

Abstract
Cancer cells optimize nutrient utilization to supply energetic and biosynthetic pathways. This metabolic process also includes redox maintenance and epigenetic regulation through nucleic acid and protein methylation, which enhance tumorigenicity and clinical resistance. However, less is known about how cancer cells exhibit metabolic flexibility to sustain cell growth and survival from low nutrient microenvironment. Here, we find that serine and glycine levels were higher in low-nutrient regions of tumors in glioblastoma multiforme (GBM) patients than they were in other regions. Metabolic and functional studies in malignant glioma cells demonstrated that serine availability and one-carbon metabolism support glioma cell survival following glutamine deprivation. Serine synthesis was mediated through autophagy rather than glycolysis. Gene expression analysis identified upregulation of methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) to regulate one-carbon metabolism in malignant glioma cells and GBM patient-derived sphere cells. In clinical samples, MTHFD2 expression was highest in the nutrient-poor areas around “pseudopalisading necrosis.” Genetic and pharmacological suppression of MTHFD2 caused tumor cell death and growth inhibition of glioma cells upon glutamine deprivation as well as autophagy inhibition did. These results may have important implications for serine-dependent one-carbon metabolism for glioma cells to survive glutamine starvation and suggest a new therapeutic strategy for patients with malignant glioma.
TMIC-22. INTERPLAY BETWEEN GBM AND IMMUNE CELLS IN THE MOUSE GLIOMA MICROENVIRONMENT AT SINGLE-CELL RESOLUTION
Authors: Lenkiewicz Anna , Jacek Karol , Ghosh Mitrajit , Cyranowski Salwador , Pilanc Paulina , Szadkowska Paulina , Ochocka Natalia , Kaminska Bozena 
Published: November 2022

Abstract
Glioblastoma (GBM) is the most common primary tumor of the central nervous system in adults. GBM patients have a particularly poor prognosis and extremely short survival time due to lack of effective therapies and rapid tumor relapse. Neoplastic cells should be effectively recognized and destroyed by the immune system, but its antitumor activity is often inhibited by tumor-secreted factors that contribute to the tumor immunosuppressive microenvironment (TME). As TME plays a key role in cancer progression and immune evasion, understanding the interplay between GBM cells and myeloid and lymphoid populations is pivotal in creating new therapeutic strategies for GBM patients. To identify subtypes and functional diversity of immune cells in glioma TME we employed single-cell RNA and protein sequencing (CITEseq, Cellular Indexing of Transcriptomes and Epitopes by Sequencing) and Visium (10XGenomics) spatial transcriptomics. We characterized the populations of myelo- and lymphoid cells, and examined their unique transcription profiles, functional diversity and localization in TME. By combining analysis of CITE-seq with spatial transcriptomics we characterized and described 35 phenotypes of immune cells, which then we localized spatially within TME in GL261 mouse gliomas. Our results indicated that the peripheral monocytes/macrophages (Cd49d) localized in the tumor core, while microglia (Tmem119) accumulated at the invasive edge. Moreover, we observed a ring of activated astrocytes and rare T lymphocytes dispersed around the tumor. Finally, Ligand-Receptor and CellChat analysis of our CITE-seq results revealed the interplay between GBM, myeloid cells and lymphocytes, indicated on potential factors responsible for accumulation and tumor-evoked reprograming of immune cells. Single-cell technologies provide high-resolution insights into cellular and functional heterogeneity of gliomas, the analysis of which, in the future, will provide us with new therapeutic strategies for GBM patients. Studies were supported by NSC grant 2020/39/B/NZ4/02683 (BK) and PACIFIC Call 1 PAS (MG).
TMIC-25. SPATIAL TRANSCRIPTOMIC LANDSCAPE OF DIFFUSE GLIOMA
Authors: Wu Zhichao , Dazelle Karen , Chung Hye-Jung , Aldape Kenneth 
Published: November 2022

Abstract
Single-cell studies have suggested heterogeneity of neoplastic cell states in diffuse gliomas. IDH-mutant astrocytoma and oligodendrogliomas (IDH-glioma) is described by cell states resembling including oligodendrocytes(OC), astrocytes (AC), and cycling neural progenitor cells (NPC); and IDH-wildtype glioblastoma (GBM-IDHwt) is marked by cell states resembling AC, NPC, oligodendrocyte progenitors (OPC) and mesenchymal (MES). Collective experience indicates that cell states are dynamic and can transition, at times recapturing brain development and inflammatory wound response. However, the spatial architecture of cell states, cell types and their interactions are yet to be investigated. We quantified the transcriptomics of 1,941 regions of interest (ROIs) from 19 IDH-glioma (non-1p/19q-codeleted n = 11, 1p/19q-codeleted n = 8) and 11 GBM-IDHwt tumors using the NanoString GeoMx Cancer Transcriptome Atlas (CTA, n = 1,811 genes) to study their spatial heterogeneity. Where appropriate, ROIs were selected to represent tumor core, infiltration edge, pseudopalisading necrosis, microvascular proliferation, and histological normal areas, if available. CTA profiling showed strong inter- and intra-tumor heterogeneity. Genes involved in oncogenesis and extra-cellular matrix such as VEGFA and VCAN, and macrophages (including CD74 and CCL3/1) showed the highest spatially variation. We identified substantial tumor immune environmental differences across ROIs including immune code, monocyte-like rich and lymphocyte-rich groups. The GBM-IDHwt cell states OPC and NPC were highly positively correlated (p-value = 5.65e-122) across ROIs suggesting spatial co-localization, but MES cell states were negatively correlated with the other three GBM cell states. M0 macrophage abundance was positively correlated with the MES state but negatively correlated with other cell states in GBM-IDHwt. The MES state was significantly lower in infiltrating edge of the tumor (p-value = 1.39e-34) but higher in the pseudopalisading necrosis region (p-value = 7.66e-30). Overall, this spatial transcriptomics dataset provides spatially informed relationships of neoplastic cell states and the immune microenvironment in diffuse glioma.
TMIC-39. TARGETING EXTRACELLULAR MATRIX HYALURONIC ACID-CD44 SIGNALING REDUCES TUMOR STEMNESS AND SENSITIZES TUMOR TO VIROTHERAPY AND ENHANCES THERAPEUTIC POTENTIAL FOR CANCER TREATMENT
Authors: Sahu Upasana , Mullarkey Matthew P , Hong Evan , Guangsheng Pei , Zhongming Zhao , Hong Bangxing , Kaur Balveen 
Published: November 2022

Abstract
The tumor microenvironment (TME), including the non-tumor cells and the extracellular matrix (ECM) plays a crucial role in tumor progression and metastasis. Hyaluronic acid (HA), the major glycosaminoglycan present in brain ECM, has long been associated with the progression and invasiveness of brain tumors. HA signals primarily thorough CD44, an adhesion/homing receptor leading to the induction of cellular AKT, MEK and HIF signaling, thereby promoting tumor cell proliferation, aggressiveness and therapy resistance. While HA in the ECM has been shown to interfere with infection and spread of oncolytic viruses, the impact of tumor-ECM interaction induced signaling on oncolytic virotherapy is heavily understudied. RNA sequencing and gene set enrichment analysis of glioma cells infected with an oncolytic Herpes Simplex Virus-1 (oHSV) demonstrate an enrichment of pathways related to tumor-ECM interaction. Immunostaining of brain sections from intracranial tumor bearing mice also reveals increased HA after oHSV treatment. Our results further demonstrate that HA/CD44-mediated signaling inhibits virus replication in vitro. Herein, to evaluate the impact of blocking tumor-ECM interactions without altering the secreted ECM, we created oHSV-sCD44, an oHSV that encodes for extracellular soluble CD44 (sCD44) that can function as a dominant negative receptor for membrane bound CD44. oHSV-sCD44 significantly reduces the stemness of glioblastoma stem cells (GSCs), induces DNA damage and sensitizes the GSCs to radiation therapy. Intra-tumoral injection of oHSV-sCD44 into patient-derived primary GBM xenograft model significantly inhibits tumor growth accompanied by reduced stemness and decreased HA expression, and increased oHSV replication and tumor cell lysis in TME. Moreover, blocking HA-CD44 signaling with a single dose of oHSV-sCD44 in murine glioma syngeneic model induces the development of a significant anti-tumor immune response with enhanced T cell infiltration. Collectively, our findings implicate oHSV-sCD44 as a potential oncolytic and immune-stimulating anticancer therapeutic.
TMIC-42. LEVERAGING B CELL IMMUNITY TO PROMOTE IMMUNOTHERAPY IN GLIOBLASTOMA
Authors: Hou David , Castro Brandyn , Zolp Andrew , Dapash Mark , Arrieta Victor , Zhao Junfei , Zhang Peng , Rashidi Aida , Burnham Emma , Patel Ronit , Lesniak Maciej , Miska Jason , Lee-Chang Catalina 
Published: November 2022

Abstract
Immunotherapy has revolutionized cancer treatment but has yet to be translated into brain tumors. Studies in melanoma and sarcoma, amongst other models, have revealed the accumulation of germinal-center-like B cells as a key survival predictor post-PD1 blockade. We seek to leverage B cell immunity to enhance immunotherapy effectiveness in glioblastoma (GBM). In human GBM and murine glioma models, we found that B cells in the tumor microenvironment (TME) are activated, but the expression of co-inhibitory molecules such as CD32 and CD72 blocks downstream effector function. Transcriptomic analysis showed high expression of inhibitory TGFβ receptors on B cells and high levels of TGFβ1 cytokine in the TME. We showed direct inhibition of B cell function through TGFβ signaling that could be prevented with TGFβ receptor blockade. Spatial multiplex immunofluorescence analysis of the TME revealed that tumor and myeloid cells express high levels of TGFβ and are also near B cells, allowing for TGFβ-mediated B cell inhibition. Blocking the TGFβ pathway via transgenic mice with TGFβ receptor knockout on B cells or TGFβ cytokine knockouts in myeloid cells, or generation of a CT2A tumor line with TGFβ cytokine knockdown, all demonstrated a survival benefit and more germinal-center-like B cells. There was also increased T cell proliferation and anti-tumor cytotoxicity. Finally, inhibiting αVβ8 integrin, a required factor that releases active TGFβ, is a translatable approach that also increased B cell proliferation and animal survival. Dual treatment with αVβ8+PD1 blockade showed the most potent survival as well as immunological memory against tumor re-challenge. Analysis of the B and T cell compartments after dual treatment showed synergy, with robust cellular proliferation and functional differentiation of plasmablasts and effector T cells. Collectively, our study highlights the importance of B cells in the TME and a remodeled approach to boost the effects of immunotherapy against GBM.
TMIC-49. CHITINASE-3-LIKE 1 PROTEIN COMPLEXES REGULATE PD-1 SIGNALING-MEDIATED IMMUNOSUPPRESSION IN GLIOBLASTOMA
Authors: Srivastava Rashmi , Chen Apeng , Dodda Meghana , Zou Han , Hu Baoli 
Published: November 2022

Abstract
Glioblastoma (GBM), the most common and lethal brain tumor with a median survival rate of only 15 months, remains largely incurable despite intensive multimodal treatment, including immunotherapeutic strategies being tested in clinical trials. GBM is highly immunosuppressive and resistant to immunotherapy because glioma cells escape from effective antitumor immunity through programing the tumor microenvironment (TME). Owing to the tremendous heterogeneity and plasticity of tumor cells and the surrounding TME, understanding the mechanisms of immune evasion by GBM remains elusive. We have recently discovered that the Chitinase-3-like-1 (CHI3L1)-Galectin-3 (Gal3) protein binding complex can selectively promote tumor-associated macrophage migration and infiltration with a protumor M2-like phenotype and T cell-mediated immunosuppression, which are governed by a transcriptional program of NF-κB/CEBPβ in the CHI3L1/Gal3-PI3K/AKT/mTOR axis. The immunoprecipitation coupled to liquid chromatography-mass spectrometry analysis revealed that galectin 3–binding protein (Gal3BP) competes with Gal3 to bind with CHI3L1 for negative regulation of the CHI3L1-Gal3 mediated processes. Interestingly, a newly-developed Gal3BP mimetic peptide can disrupt CHI3L1-Gal3 interaction, resulting in decreasing migration of M2-like bone marrow-derived macrophages (BMDMs), increasing CD8+ T cell infiltration, reversing immunosuppression, and inhibiting tumor progression in vitro and in vivo. Analyzing PD-1 signaling activation, we found that the Gal3BP mimetic peptide significantly decreased PD-L1 expression in tumor cells. Correlation analysis showed that CHI31L and Gal3 (encoded by LGALS3 gene) are significantly associated with both PD-L1 and PD-L2 in GBM patient samples. Furthermore, overexpression of CHI3L1 increased expression levels of PD-L1 and PD-L2, and CHI3L1 deletion decreased their expression in GBM patient-derived neurosphere lines. The treatment with recombinant CHI3L1 protein significantly increased PD-L1 and PD-L2 expression in M2-like BMDMs (with high levels of endogenous Gal3). Collectively, these data suggest that CHI3L1 protein complexes control the GBM immunosuppressive microenvironment by PD-1/PD-L1/PD-L2 signaling, providing new immunotherapeutic strategies for this brain cancer.
TMIC-52. RELATIONSHIP BETWEEN MACROPHAGE AND RADIOSENSITIVITY IN HUMAN PRIMARY AND RECURRENT GLIOBLASTOMA: IN SILICO ANALYSIS WITH PUBLICLY AVAILABLE DATASETS
Authors: Kim In Ah , Jang Bum Sup 
Published: November 2022

Abstract
The glioblastoma microenvironment predominantly contains tumor-associated macrophages that support tumor growth and invasion. We investigated the relationship between tumor radiosensitivity and infiltrating M1/M2 macrophage profiles in public datasets of primary and recurrent glioblastoma. We estimated the radiosensitivity index (RSI) score based on gene expression rankings. Macrophages were profiled using the deconvolution algorithm CIBERSORTx. Samples from The Cancer Genome Atlas (TCGA), Chinese Glioma Genome Atlas (CGGA), the Ivy Glioblastoma Atlas Project dataset, a single-cell RNA sequencing dataset (GSE84465), Glioma Longitudinal Analysis Consortium (GLASS), and an immunotherapy trial dataset (GSE121810) were included. RSI-high radioresistant tumors were associated with worse overall survival in TCGA and CGGA than RSI-low tumors. M1/M2 macrophage ratios and RSI scores were inversely associated, indicating that radioresistant glioblastoma tumor microenvironments contain more M2 than M1 macrophages. In the single-cell RNA sequencing dataset, the mean RSI of neoplastic cells was positively correlated with high M2 macrophages proportions. A favorable response to programmed cell death protein 1 (PD-1) therapy was observed in recurrent glioblastomas with high M1/M2 macrophage ratios and low RSI scores. In patients with recurrent glioblastoma, fewer M2 macrophages and low RSI scores were associated with improved overall survival. High M2 macrophage proportions may be involved in radioresistant glioblastoma.