bluuebunny/crossref_metadata_2025_split huggingface dataset:
EXTH-56. TARGETING GBM WITH AN ONCOLYTIC PICORNAVIRUS SVV-001 ALONE AND IN COMBINATION WITH FRACTIONATED RADIATION IN A NOVEL PANEL OF ORTHOTOPIC PDX MODELS
Authors: Zhang Huiyuan , Du Yuchen , Qi Lin , Xiao Sophie , Braun Frank K , Kogiso Mari , Abdallah Aalaa , Baxter Patricia A , Su Jack M , Brat Daniel , Hallenbeck Paul L , Teo Wan-Yee , Patel Akash , Li Xiao-Nan 
Published: November 2022

Abstract

BACKGROUND
Large panels of clinically relevant animal models are needed for the development of new therapies for GBM. SVV-001, a replication-competent oncolytic virus that can pass through the blood brain barrier, represents an attractive novel approach. Material and

METHODS
Surgical tissues from 23 patients were implanted into the brains of NOD/SCID mice (1x105 cells/mouse) to develop patient-derived orthotopic xenograft (PDOX) models. Analysis of histology, gene expression (RNAseq), tumor growth was performed and compared with the patient tumors during subtransplantations (up to 5 passages). Anti-tumor activities of SVV-001 were examined in vitro in primary monolayer cultures, 3D neurospheres and purified glioma stem cells derived from 13 PDOX models; and its therapeutic efficacy validated in vivo in PDOX models through single i.v. injection (1x1011 viral particle) alone and in combination with fractionated (2 Gy/day x 5 days) radiation. Changes of animal survival times were analyzed together with viral infection, cell proliferation and DNA damage.

RESULTS
PDOX formation was confirmed in 17/23 (73.9%) GBMs. They replicated key histopathological features and diffuse invasion of the patient tumors. RNAseq identified the differentially expressed genes and subclassified PDOX models into proneural, classic and mesenchymal groups. SVV-001 killed primary monolayer cultures (4/13 models) and neurosphere (7/13 models) and putative glioma stem cells (CD133+ and/or CD15+ cells) in vitro, infected PDOX tumors without harming normal brain cells in vivo and significantly prolonged survival times in 2/2 models. Combination with radiation showed a trend in further prolonging animal survival times through enhanced DNA damages by SVV-001.

CONCLUSION
A panel of 17 clinically relevant and molecularly annotated PDOX modes of GBM is developed. The strong anti-tumor activities of SVV-001 in vitro and in vivo in GBM models provided experimental rational that support the initiation of clinical trials in GBM patients.
EXTH-62. EXAMINATION OF THE TUMOR MICROENVIRONMENT OF GLIOMAS TREATED WITH VIROIMMUNOTHERAPY REVEALS THE PRODUCTION OF ONCOMETABOLITES LEADING TO IMPROVED THERAPEUTIC STRATEGIES
Authors: Nguyen Teresa , Shin Dong Ho , Kim Debora , Singh Sanjay K , Jiang Hong , Fan Xuejun , Yi Yanhua , Gumin Joy , Lang Frederick , Alvarez-Breckenridge Christopher , Alonso Marta M , Zhai Lijie , Ladomersky Erik , Lauing Kristen , Wainwright Derek , Fueyo Juan , Gomez-Manzano Candelaria 
Published: November 2022

Abstract
Oncolytic viruses are considered part of immunotherapy and have shown promise in preclinical experiments and clinical trials in a small fraction of patients. To improve the response rate, it is necessary to reshape the tumor microenvironment that shields the tumor from the immune system of the patient. We engineered Delta-24-RGDOX (DNX-2440), an oncolytic adenovirus to express the T-cell activator OX40L, which triggered, in murine glioblastoma models, a dramatic reshaping of the tumor microenvironment dominated by changes in immune processes in an RNA-seq analysis of ingenuity pathways. Paradoxically, network analyses also revealed that Delta-24-RGDOX induced robust activation of the cytokine-driven immunosuppressive IDO-Kynurenine-AhR circuitry, indicating a potential mechanism of resistance of gliomas to oncolytic virotherapy. To reverse this immunosuppression, we combined Delta-24-RGDOX with clinically relevant IDO inhibitors to treat glioma-bearing mice. The combined therapy decreased the activation of the IDO network without affecting viral activity. Flow cytometry assays revealed a decrease in the presence of the immunosuppressive cell populations, MDSCs and Tregs, which presence was higher in Delta-24-RGDOX-treated tumors. These data were further confirmed in gene set enrichment analyses. Functional co-culture studies showed that the combined therapy activated splenocytes against tumor antigens and that this activation was reversed by Kynurenine. Importantly, the combination treatment eradicated the tumors in a CD4-dependent manner and significantly prolonged the survival of glioma-bearing mice. Altogether, these studies indicate that the combination treatment promotes an adaptive immune response while decreasing the immunosuppression caused by the virus-induced IDO activation. Furthermore, our data identified the striking role of immunosuppressive pathways in the resistance of gliomas to oncolytic virotherapy. Specifically, the activity of the tumor microenvironment IDO circuitry was responsible, at least partially, for the remodeling of local immunosuppression after tumor infection. Combining molecular and immune-related therapies may improve outcomes in human gliomas treated with virotherapy.
EXTH-64. IPSC-DERIVED NK CELLS EXHIBIT POTENT IN VITRO AND IN VIVO TUMORCIDAL ACTIVITY AGAINST PATIENT-DERIVED GLIOBLASTOMA STEM CELLS (GSCS)
Authors: Ning Jianfang , Davis Zachary , Cichocki Frank , Wang Hongbo , Tuininga Katie , Bjordahl Ryan , Lee Tom , Miller Jeffrey , Chen Clark 
Published: November 2022

Abstract

INTRODUCTION
Natural Killer (NK) cells play pivotal roles in cancer immune surveillance and harbor potent anti-tumor activities. We have engineered an induced pluripotent stem cells (iPSCs) derived, NK cell (termed FT538) that express 1) a high-affinity non-cleavable CD16 Fc receptor to facilitate antibody engagement, 2) a membrane-bound IL-15/IL-15 receptor fusion to enhance NK activity and persistence and 3) a knock-out of CD38 to enhance NK metabolic fitness. FT538 is produced in a GMP-compliant manner as an off-the-shelf immunotherapy. Clinical experiences with FT538 have shown evidence of durable anti-tumor immunity after single administration in select lymphoma patients.

METHODS
We performed pre-clinical studies to characterize the tumoricidal activity of FT573 against glioblastomas.

RESULTS
Without specific antibody to engage CD16, FT538 exhibited potent in vitro activity against ~80% of a panel of isocitrate dehydrogenase wild-type patient-derived xenograft (PDX) glioblastoma lines, including MGG8. When directly injected into the murine brain, 50% of the FT538 persisted for &gt; 2 weeks without addition of exogenous cytokine. Moreover, the mice exhibited no evidence of neurologic compromise, and brains harvested after injection show no signs of FT538 activation. In vivo intratumoral injection of FT538 into an orthotopically established MGG8 glioblastoma caused a ~20 fold increase in granzyme B release by FT538, with a comparable increase of cleaved-caspase 3 in the tumor. In independent experiments, &gt; 90% of mice orthotopically implanted MGG8 survived beyond 100 days after intra-tumoral FT538 while all mice treated with mock injection died by 60 days. For glioblastoma PDX refractory to FT538, such as MGG4, tumoricidal activity of FT583 can be induced in vitro and in vivo by the addition of a tri-specific engager targeting B7H3, a surface protein highly expressed in glioblastomas.

CONCLUSION
Our results demonstrate great promise of FT538 as a glioblastoma therapy, with plans for translation into a first-in-human clinical trial.
EXTH-66. BIOENERGETIC SWITCH AND BIOLOGIC RESPONSE WITH SLC25A11 INHIBITION TARGETING MALATE-ASPARTATE SHUTTLE (MAS) ON GLIOBLASTOMA TUMORSPHERE
Authors: Cho Hejoung , Yoo Jihwan , Choi Ran Joo , Park Junseong , Yoon Seon-Jin , Oh Yoojung , Moon Ju Hyung , Kim Eui Hyun , Chang Jong Hee , Kang Seok-Gu 
Published: November 2022

Abstract
BACKGROUND: Cancer cells depended on cytosolic nicotinamide adenine dinucleotide (NADH) transported into mitochondria via the malate-aspartate shuttle (MAS) for ATP production. KN612 (N-phenylmaleimide), a MAS inhibitor, is known to interfere with cancer growth by reducing ATP production, supported by several studies except on glioblastoma (GBM). Therefore, this study was designed to elucidate whether MAS could be an aimable target in GBM. METHODS: We compared expression levels of MAS conforming enzymes between normal and GBM samples. Gene expression profiles were analyzed using RNA-sequencing. Mitochondrial activity was measured by oxygen consumption (OCR), tetramethylrhodamine-ethylester (TMRE) staining, and liquid chromatograph-tandem mass spectrometer (LC-MS/MS). Also, biological functions were measured by cell viability, ATP levels, NADH levels, stemness, and invasiveness. In vivo efficacies were confirmed using a mouse orthotopic xenograft model. RESULTS: An analysis of the microarray database revealed that expression levels of several MAS enzymes including OGC (SLC25A11) were elevated in GBM. Through RNA sequencing, it was confirmed that KN612 accurately targeted SLC25A11 and decreased its expression. KN612-treated cells showed decreased viability, ATP production, and NADH levels compared with control cells. Under the same conditions, a significant decrease in stemness, invasion, and MMP was confirmed. In addition, KN612 confirmed remarkable therapeutic responses in a mouse orthotopic xenograft model. CONCLUSION: Our results show that KN612 effectively inhibits cancer cells both at the cellular level and at the in vivo level. This shows that targeting MAS could be a potential treatment option in addition to the currently limited standard GBM therapy.
EXTH-71. CHEMOTACTIC RECOMBINANT ADENO-ASSOCIATED VIRUS (RAAV) VIROTHERAPY IMPROVES LYMPHOCYTE RECRUITMENT IN GLIOBLASTOMA (GBM)
Authors: von Roemeling Christina , Klippel Kelena , Trivedi Vrunda , Yegorov Oleg , Yang Changlin , Hoang-Minh Lan , Doonan Bently , Ran Yong , Golde Todd , Mitchell Duane 
Published: November 2022

Abstract

BACKGROUND
Although immunotherapy holds promise for producing durable outcomes in GBM, a major obstacle is the limited ability of cytotoxic T lymphocytes (CTLs) to home to the tumor. CTLs are recruited via long-range signaling mediated by the diffusion of chemokines present in inflammatory environments. During glioma formation, tumors manufacture immune-suppressive chemokines and cytokines that co-opt resident cells, resulting in the preferential recruitment of immune suppressor cells from the periphery. We explored a novel strategy to selectively reprogram the tumor microenvironment (TME) using focal delivery of rAAV encoding a powerful call-and-receive chemokine for CTLs: CXCL9. HYPOTHESIS: rAAV transduction of the GBM TME to express CXCL9 (rAAV6-CXCL9) will enhance CTL tumor infiltration and improve responsiveness to immunotherapy.

METHODS
Proteomic arrays were used to identify CTL chemokines absent in human glioma. 3D immunohistochemistry and flow cytometry was used to evaluate geospatial rAAV transgene expression and lymphocyte trafficking in vitro and in vivo. Survival studies of rAAV6-CXCL9 alone and in combination with anti-PD-1 checkpoint blockade was performed in preclinical models of GBM. Tumor immunogenicity following treatment was evaluated by scRNAseq and proteomic analysis.

RESULTS
We identify rAAV6 as a reliable capsid for transducing murine tumor-reactive astrocytes. Transgene expression following a single intra-tumor injection is focal, stable, and results in high levels of CXCL9 production. CXCL9 is effective at promoting tumor T-cell chemotaxis in vitro and in vivo. rAAV6-CXCL9 treatment sensitizes GBM to PD-1 blockade, improving survival in two distinct syngeneic preclinical models- an effect that is largely dependent on CD8 T-cells.

CONCLUSIONS
We combine the excellent safety profile of rAAV with focal delivery, enhancing exposure within the tumor compartment and mitigating systemic toxicities seen with conventional therapies. rAAV therapy was successful at stimulating the intended biological response to our encoded transgene (CXCL9): increased lymphocyte recruitment. This strategy has far-reaching potential across other immunotherapy platforms.
EXTH-72. TTFIELDS ENHANCE THE ANTINEOPLASTIC ACTIVITY OF THE DRUG-REPURPOSING APPROACH CUSP9V3 IN GLIOBLASTOMA IN VITRO
Authors: Cao Qiyu , Hajosch Annika , Kast Richard , Hlavac Michal , Westhoff Mike-Andrew , Siegelin Markus , Wirtz Christian Rainer , Halatsch Marc-Eric , Karpel-Massler Georg 
Published: November 2022

Abstract

OBJECTIVE
Drug repurposing represents a promising strategy to safely accelerate the clinical use of therapeutics with antineoplastic activity. In this study, we examined whether Tumor Treating Fields (TTFields) enhance the biological effects of CUSP9v3, a treatment strategy including nine repurposed drugs, in an in vitro setting of glioblastoma.

METHODS
We performed MTT-assays to examine effects of the combination treatment on the viability of different glioblastoma cells. Tumor spheroids were used as a model to examine effects of the combination treatment in a 3-dimensional setting. Staining with annexin V/propidium iodide or MitoTrackerTM followed by flow cytometry was done to assess pro-apoptotic effects. Specific protein expression of caspases and members of the Bcl-2 family of proteins was determined by Western blot analyses.

RESULTS
TTFields had at least additive inhibitory effects on the cell viability of established (U251), primary cultured (PC38, PC40, PC128) and stem-like (SC38, SC40) glioblastoma cells when combined with CUSP9v3. In addition, flow cytometric analyses revealed that a simultaneous treatment with TTFields and CUSP9v3 significantly increased the fraction of annexin V-positive (apoptotic) glioblastoma cells. Moreover, the fraction of cells with a reduced mitochondrial outer membrane potential was significantly higher following a simultaneous treatment with TTFields and CUSP9v3. On the molecular level, these observations were associated with enhanced cleavage of effector caspase 3 and a reduced expression of the anti-apoptotic Bcl-2 family proteins Bcl-2 and Mcl-1.

CONCLUSION
These data suggest that TTFields enhance the susceptibility of glioblastoma cells towards CUSP9v3, potentially allowing significant dose reduction and decreased toxicity. This observation seems to rely at least in part on a caspase-dependent cell death mechanism. TTFields are widely used for the treatment of glioblastoma patients and CUSP9v3 was recently shown to have a favorable safety profile in a phase Ib/IIa trial (NCT02770378) which facilitates transition of a combined approach to the clinical setting.
EXTH-73. A NOVEL ANTI-GALECTIN-3 MONOCLONAL ANTIBODY THERAPEUTIC DIMINISHES MIGRATION AND SLOWS TUMOR GROWTH TO PROLONG SURVIVAL IN GLIOBLASTOMA
Authors: Barrette Anne Marie , Deshpande Krutika , Cadiente Marvin , Wang Xi , Sun Dongxu 
Published: November 2022

Abstract
Glioblastoma (GBM) is the most aggressive, invasive, and malignant primary brain cancer with a standard of care that hasn’t been improved in over 15 years. Heterogenous tumor cell populations that evade resection and develop resistance to chemoradiation highlight the need for novel therapies that can target both the migratory and proliferative nature of GBM cells. Galectin-3, a carbohydrate-binding protein overexpressed in the GBM parenchyma and stromal astrocytes and microglia/macrophages, is a potent modulator of cell proliferation, migration, angiogenesis, T-cell inhibition, and M2 macrophage polarization. Increased Galectin-3 expression levels correlate to lower survival in glioma patients, and these levels further increase following exposure to temozolomide or radiation alone or in combination, thus more greatly emphasizing the need to target Galectin-3 in GBM patients following initial standard-of-care treatment. Here, we characterize the therapeutic effects of a monoclonal anti-Galectin-3 antibody (Gal3 Ab) we have developed for its ability to inhibit GBM tumorigenic and metastatic potential in several in vitro assays and in vivo heterotopic and orthotopic mouse models. We find that Galectin-3 directly binds to EGFR, the most commonly amplified gene locus in GBM affecting roughly 40% of patients, and our Gal3 Ab can block this binding. Human GBM cells treated with Gal3 Ab show a significant decrease in migration and invasion and reduction in EGFR activation in vitro. In vivo, the Gal3 Ab additionally decreases tumor burden and provides a survival benefit in combination with temozolomide compared to the current standard-of-care treatment alone. Our studies indicate the potential for our novel anti-Galectin-3 antibody to further benefit GBM patients receiving chemoradiation treatment by diminishing tumor cell migration and slowing tumor growth.
EXTH-81. ITGA5 IS A NOVEL IMMUNOTHERAPEUTIC TARGET AGAINST TREATMENT REFRACTORY MEDULLOBLASTOMA
Authors: Bakhshinyan David , Suk Yujin , Kuhlman Laura , Adile Ashley , Ignatchenko Vladimir , Gwynne William , Custers Stefan , Macklin Andrew , Venugopal Chitra , Kislinger Thomas , Singh Sheila 
Published: November 2022

Abstract
Medulloblastoma (MB) is the most common type of malignant pediatric brain cancer. Current standard of care (SOC) involves maximal safe resection and neuraxis radiotherapy and chemotherapy in individuals older than 3 years. To date, these cytotoxic SOC combined with craniospinal irradiation led to devastating neurocognitive and developmental deficits impacting quality of life for pediatric patients. The biological heterogeneity of MB is highlighted by the existence of four distinct molecular subgroups (WNT, SHH, Group 3, and Group 4). Group 3 and Group 4 have the poorest patient outcomes because of their aggressive, metastatic nature, and so often remain treatment refractory to SOC. Group 3 has a poor prognosis due to its high incidence of leptomeningeal spread and an overall survival rate of less than 50%. The cytotoxic nature and lack of response in specific subtypes to SOC underscores the urgent need for developing and translating novel treatment options including immunotherapies. In our earlier work, we have developed a therapy-adapted patient derived xenograft (PDX) model of the Group 3 MB as the tumor cells undergoes therapy in vitro and in vivo. N-glycocapture surfaceome profiling of the MB cells through this PDX model identified Integrin α5 (ITGA5) as one of the most differentially expressed targets found at recurrence when compared to engraftment and untreated timepoints. Through shRNA knockdown and small molecule inhibition, we identify ITGA5 expression marks a MB cell subpopulation with increased self-renewal ability both in vitro and in vivo. Access to recurrent MB (rMB) post-therapy allowed us to investigate the changes in the surfaceome of MB cells using proteomics profiling to identify promising rMB-specific targets for rational development of novel immunotherapies.
EXTH-83. DEVELOPMENTALLY REGULATED ANTIGENS FOR IMMUNOLOGIC TARGETING OF EMBRYONAL PEDIATRIC BRAIN CANCERS
Authors: Sebastian Mathew , Pham Christina , Wummer Brandon , Woodworth Delaney , Reid Alexandra , Abraham Rebecca , Gil Jorge , Drake Jeffrey , Mitchell Duane , Flores Catherine 
Published: November 2022

Abstract

BACKGROUND
Mutated neoantigens have shown promise as targets for cancer immunotherapy but embryonal pediatric brain tumors with notoriously low mutational burden have more limited opportunities for neoantigen directed therapies. Evidence suggests that embryonal brain tumors such as medulloblastoma (MB) and brain stem gliomas (BSG) arise from the aberrant reactivation of fetal developmental programming. We explored the hypothesis that neonatal mouse cerebellum and brainstem express developmentally regulated proteins that could serve as potent tumor rejection antigens for a preclinical model of Group 3 MB (NSC) and H3.3K27M positive BSG (K2), respectively. We thus generated tumor-reactive T cells by using P5 cerebellum and P4 brain stem RNA as sources of antigens. P5-cerebellum-specific T cells (cDevAg-T cells) and P4-brain stem-specific T cells (bsDevAg-T cells) demonstrated effector function against respective MB and BSG tumor cells with exquisite specificity.

METHODS
RNA was isolated from P5 cerebellum and P4 brain stem and pulsed into bone marrow-derived dendritic cells. These were then used to in vitro activate splenocytes from previously immunized mice, generating either cDevAg-T cells or bsDevAg-T cells. In vitro functionality assays against tumor targets were conducted to determine reactivity and specificity. DevAg-T cells were used in adoptive cellular therapy in orthotopic models to determine therapeutic efficacy of DevAg-specific T cells in vivo. Results and

CONCLUSIONS
DevAg-T cells produce high levels of Th1-type cytokines that recognize distinct subtypes of MB and BSG, show no cross-reactivity with normal brain. Adoptive cellular therapy employing DevAg-specific T cells demonstrate a significant survival benefit in orthotopic models of established Group 3 NSC MB and H3.3K27M mutation positive BSG. Our studies demonstrate that RNA encoding non-mutated and organ-specific developmental antigens can serve as novel tumor rejection antigens for pediatric brain cancers.
EXTH-88. DOCETAXEL TARGETS AGGRESSIVE METHYLATION PROFILES AND SERVES AS A RADIOSENSITIZER IN HIGH-RISK MENINGIOMAS
Authors: Youngblood Mark , Tran Anh , Wang Wenxia , An Shejuan , Scholtens Denise , Zhang Lyndsee , O’Shea Kaitlyn , Pokorny Jenny , Magill Stephen , Sachdev Sean , Lukas Rimas , Ahmed Atique , Unruh Dusten , Walshon Jordain , McCortney Kathleen , Wang Yufen , Baran Aneta , Sahm Felix , Aldape Kenneth , Chandler James , Heimberger Amy , James David , Horbinski Craig 
Published: November 2022

Abstract
DNA methylation profiling has emerged as a transformative prognostic tool in meningiomas, identifying distinct epigenetic subgroups that share unique clinical and biological features. Hypothesizing that methylation patterns may also reflect underlying pathogenic mechanisms, we investigated if this modality could be informative for the selection of efficacious chemotherapies in meningiomas, focusing on FDA-approved compounds that are well-tolerated in patients. Using a methylation dataset of 210 tumors, we identified 981 genes in which adjacent CpG sites were predictive of progression-free survival. Pathway enrichment of these genes nominated Docetaxel as a candidate therapeutic; a well-established taxane that is routinely used for breast, prostate, and lung cancers. Using 15 primary patient-derived cultures and 2 cell lines, we found that Docetaxel could inhibit meningioma growth at concentrations that were favorable relative to other cancers. This drug triggered apoptosis in meningiomas of all tumor grades and methylation classes, with IC50 values ranging from 0.8nm to 4.4nm in vitro. Combining Docetaxel with radiation therapy resulted in synergistic cytotoxic effects on meningioma cultures, associated with elevated markers of cell death and dsDNA breaks. We confirmed our results via orthotopic meningioma mouse models of the cell lines CH157 and IOMM-Lee, which exhibited dose-dependent improvements in mortality and tumor volume in response to Docetaxel and radiation. Our results leverage DNA methylation patterns to repurpose FDA-approved medications in the treatment of meningiomas, and suggest a clinical trial of Docetaxel with radiotherapy may be efficacious in higher-grade lesions.