bluuebunny/crossref_metadata_2025_split huggingface dataset:
TMIC-17. ASTROCYTE SENESCENCE CONTRIBUTES TO SEX DIFFERENCES IN THE AGE-RELATED INCREASE IN GLIOBLASTOMA INCIDENCE
Authors: Broestl Lauren , Sponagel Jasmin , Rhee Gina , Grandison Lucia , Kfoury Najla , Rockwell Nathan , Hill Cameron , Rubin Joshua 
Published: November 2019

Abstract
Incidence rates for glioblastoma increase exponentially with age in a sex-specific manner, with a steeper increase observed in men. This suggests that age may be a more significant factor in promoting male tumors than female tumors. Drivers in the age-related cancer increase include both the accumulation of DNA damage and microenvironment changes, specifically an increase in the number of senescent cells. Cellular senescence comprises two distinct components – a cell cycle arrest program that prevents proliferation of mutated cells and acts as a potent anti-cancer mechanism, and a paracrine signaling component that can have both pro- and anti-tumorigenic properties. The contribution of senescence to brain tumor incidence and outcome is currently unknown; additionally, no research has been done on whether there are sex differences in mechanisms of senescence induction or in the secretory phenotype of senescent cells. Using both wildtype astrocytes and a glioblastoma model, we show that female cells have a lower threshold for senescence induction in response to oxidative stress, telomere shortening, and DNA damage, and that sex differences in senescence induction in response to DNA damage are in part mediated by differences in the regulation of p21. In addition, we show that male and female senescent astrocytes differ in their secretory phenotypes. Male senescent astrocytes secrete more tumor promoting factors, and conditioned media from male but not female senescent astrocytes promotes tumor cell proliferation. These findings provide a potential explanation for sex differences in glioblastoma incidence rates. Achieving a better understanding of mechanisms of senescence and how they differ in male and female cells could advance development of senescence directed therapies and potentially improve cancer treatment for a range of tumor types.
TMIC-38. CLINICAL AND IMMUNOLOGICAL CHARACTERIZATION OF HERPES VIRUS ENTRY MEDIATOR IN HUMAN GLIOMAS
Authors: Han Ming-Zhi , Wang Shuai , Wang Dong-Hai 
Published: November 2019

Abstract

BACKGROUND
Dysregulation of immune checkpoint members within tumors, including glioblastoma (GBM), is related to immune evasion. Herpes virus entry mediator (HVEM) is a novel identified immune checkpoint molecule which plays essential roles in both innate and acquired immunity. Despite recent advances in exploring the function HVEM in a variety cancer types, the clinical and immunological importance of HVEM in human gliomas remain largely unknown.

METHODS
Molecular and clinical data was obtained from publicly genomic databases. Immunohistochemistry was applied to assess the protein level of HVEM. Matlab software as well as R language were used for statistical analysis.

RESULTS
HVEM was found to be elevated in aggressive gliomas, especially in isocitrate dehydrogenase (IDH) wild-type GBM. High expression of HVEM was associated with Mesenchymal subtype and showed promising prognostic values based on Cox regression model and nomogram model. HVEM showed intra-tumor heterogeneity with abundant in peri-necrotic zone and microvascular region. In addition, HVEM high patients were more frequent with genomic aberrations of oncogenic events. Gene ontology and pathway analysis uncovered the enrichment of HVEM in multiple immune regulation process, especially in the suppression of T cell mediated immunity in GBM. Moreover, HVEM was tightly associated with several infiltrating immune and stromal cell lineages in microenvironment, and showed high correlation with other immune checkpoints.

CONCLUSIONS
Our data highlights the importance of HVEM in GBM progression and that targeting HVEM combined with current immune checkpoint blockades might be a novel therapeutic strategy for GBM.
TMIC-62. FYN, AN EFFECTOR OF ONCOGENIC RECEPTOR TYROSINE KINASES SIGNALING IN GLIOBLASTOMA, INHIBITS ANTI-GLIOMA IMMUNE RESPONSES: IMPLICATIONS FOR IMMUNOTHERAPY
Authors: Comba Andrea , J Dunn Patrick , E Argento Anna , Kadiyala Padma , Ventosa Maria , Patel Priti , Zamler Daniel , Nunez Felipe , Zhao Lili , Castro Maria , Lowenstein Pedro 
Published: November 2019

Abstract
There is currently much excitement for the use of immunotherapies in cancer. In spite of positive results using checkpoint inhibitors in melanoma and CAR T cells in leukemias, these strategies have not yet achieved robust clinical responses in human gliomas. A powerful inhibitory microenvironment is thought to be the culprit. Mechanisms that determine the inhibitory microenvironment remain poorly understood. Herein we demonstrate that FYN, a downstream target of receptor tyrosine kinases signaling, inhibits the anti-glioma immune response. We utilized genetically engineered mouse models (GEMM) of glioma based on the Sleeping Beauty Transposon method. We examined the activities of FYN in NP (N-ras + shp53), NPA (NP + shATRX), and NPD (NP + PDGF overexpression) tumors. We also generated FYN knockdown glioma stem cells in vitro to induce gliomas in immune-competent and immune-deficient mice of varying genetic backgrounds (NSG, CD8-/-). Flow cytometry was used to characterize immune cells within the glioma microenvironment. Our results show that FYN knockdown in NP, NPA, or NPD GEMM of glioma reduced tumor progression and increased survival by 25–77%. GSEA analysis of differential expressed genes of WT vs. FYN knockdown gliomas revealed enrichments of gene ontologies related to immune functions. In NSG and CD8-/- immune-deficient mice, FYN knockdown failed to inhibits tumor growth and increase animal survival. These results suggest that FYN enhances tumor progression through changes in anti-glioma immune activation. Examination of tumor immune infiltrates by flow cytometry indicate a 50–70% reduction in powerful immune inhibitory myeloid derived cells (MDSCs) with no changes in the total level of CD8+ and CD4+ cells. Our results show for the first time that FYN reduces anti-glioma immune responses, likely through a reduction in inhibitory MDSCs. The specific inhibition of FYN exclusively within glioma cells, could improve the efficacy of anti-glioma immunotherapies.
TMOD-04. CHARACTERIZATION OF MUTANT IDH1 OLIGODENDROGLIOMAS
Authors: Heinz Rick , Holmen Sheri 
Published: November 2019

Abstract
Gliomas are the most common primary central nervous system malignancy in adults, but the molecular mechanisms responsible for their development and progression are not fully understood. Recent genomic analysis of World Health Organization grade II-III gliomas identifies 3 molecular subtypes of low grade glioma: no IDH mutation; IDH mutation without 1p/19q co-deletion; and IDH mutation with 1p/19q co-deletion. The latter, categorized as oligodendroglioma, commonly expresses loss of function mutations in CIC, FUBP1, and activation of PIK3CA. However, it is unknown if any of these mutations are sufficient to promote glioma development in cooperation with mutant IDH. Furthermore, research in oligodendroglioma is hampered by the lack of in vivo models of this specific glioma subtype. By utilizing the established RCAS/TVA somatic cell gene delivery method, mutant IDH1 can be expressed in the brains of mice. The tumorgenicity of other mutated genes associated with oligodendroglioma can be determined using additional methods such as conditional gene knockout and in vivo CRISPR-Cas9 mediated deletion. A mouse model for oligodendroglioma may identify new targetable genetic drivers of oligodendroglioma that will be useful for testing novel therapeutic strategies.
TMOD-06. MODELLING ADULT AND PAEDIATRIC GLIOBLASTOMA MULTIFORME (GBM) USING A GENE SCREEN-BASED APPROACH IN MICE AND HUMAN IPSC-DERIVED CEREBRAL ORGANOIDS
Authors: Antonica Francesco , Garilli Francesca , Del Mar Gardeazabal Bataller Maria , Santomaso Lucia , Tiberi Luca 
Published: November 2019

Abstract
Glioblastoma multiforme (GBM) represent the most devastating form of high-grade glioma (HGG) affecting adults and children. Despite a multi-therapeutic approach consisting in surgery, radio- and chemo-therapy the prognosis remains poor. Several models such as xenograft, animal models and recently organoids, have been developed in order to investigate the physiopathology of GBM. Although several mouse models (where either gain- or loss-of function of genes/pathway found altered in patients induce tumour formation) have been generated many aspects of how the tumour is formed, evolves, infiltrates and recurs after treatments remain unclear. Another challenge is the creation of proper GBM models showing intratumor heterogeneity found in the patient tumour but missing in the animal model generated so far. To overcome the lack of that human tumour characteristic, we decided to generate new model of GBM using a gene screen-based approach in mice. Firstly, we analysed the genes found amplified or mutated in GBM patients; secondly, we over-expressed the candidate genes (combination of multiple genes found to be mutated or amplified in specific GBM patients) in the subventricular zone (SVZ) of P2 mouse brain. Thirdly, we analysed the formation of tumour after 2 months. We over-expressed roughly 50 combinations in newborn mice and found that only 3 successfully led to the formation of lesions positive for proliferation and brain tumour marker (i.e. GFAP). We are currently characterising the tumours by DNA methylation analysis and RNA-seq (for further confirmation of the cancer subtype and intratumor heterogeneity) or investigating the cell of origin of the tumour. Moreover, we are testing the tumorigenicity of specific in GBM-amplified/mutated gene combinations in hiPSC-derived cerebal organoids. Our data suggest that a gene screen-based approach can be used for quickly and easily assaying the tumorigenicity of genes found amplified/mutated in GBM patients as well as the biology behind such complex process.
ANGI-01. IDENTIFICATION OF INVASION-DETERMINISTIC TRANSCRIPTION FACTORS IN GLIOBLASTOMA USING INTEGRATIVE TRANSCRIPTOME ANALYSIS
Authors: Park Junseong , Shim Jin-Kyoung , Eun Lee Jae , Jin Yoon Seon , Yoo Jihwan , Hyung Moon Ju , Hyun Kim Eui , Hee Chang Jong , Kang Seok-Gu 
Published: November 2019

Abstract
Glioblastoma (GBM) is one of the most lethal human tumors with a highly infiltrative phenotype. Although invasiveness is related to poor prognosis, no therapeutic intervention targeting invasion is available for treatment of GBM patient, partially due to the diversity and redundancy of invasion machinery genes. In this regard, additional identification of deterministic and causative targets for invasion is required. Invasiveness of GBM patients and matched tumorspheres (TSs) was quantified using MR images and collagen-based 3D invasion assays, respectively. Transcriptome of GBM samples were obtained using microarrays. The knockdown effects of invasion-deterministic transcription factors (TFs) were evaluated using western blot and mouse orthotopic xenograft model. This study is aimed to identify novel transcriptional regulatory networks, which can collectively modulate invasion-involved genes in GBM. After classification of 23 GBM patient-derived TSs into low and high invasion groups, we applied single sample gene set enrichment analysis using TF target gene sets. According to enrichment scores, TFs responsible for low (PCBP1) and high (STAT3 and SRF) invasiveness were identified. Consistently with computational prediction, knockdown of PCBP1 significantly increased invasion area, whereas knockdown of STAT3 or SRF significantly suppressed invasive properties in all tested TSs. Notably, MR images showed coherent patterns with invasion of originated TS, and high invasiveness was associated with poor prognosis. In addition, mouse orthotopic xenograft model using TSs with down-regulated STAT3 or SRF showed significantly prolonged survival time compared to control. We identified invasion-deterministic TFs in glioblastoma using integrative transcriptome analysis. Owing to relationship among these transcriptional regulatory networks, invasive phenotype, and prognosis, we suggest that these TFs as novel drug targets for GBM.
TMOD-26. MODELING GLIOBLASTOMA BY IMPLANTATION OF INTACT PATIENT-DERIVED ORGANOIDS INTO RODENT BRAINS
Authors: Nguyen Phuong , Jacob Fadi , Salinas Ryan , Zhang Daniel , Song Hongjun , Ming Guo-li 
Published: November 2019

Abstract
Glioblastoma multiforme (GBM) is the most common primary and aggressive brain tumors in adults with extremely poor prognosis and limited treatment options. A major hallmark of GBM is the rapid and diffused infiltration of tumor cells into the surrounding healthy tissue that contribute to tumor recurrence and therapeutic resistance. However, existing in vitro cell culture or in vivo xenograft models inadequately recapitulate the inter-tumoral and intra-tumoral heterogeneity which are key features of GBM. For example, common oncogenic drivers of GBM such as epidermal growth factor receptor (EGFR) amplification and EGFRvIII mutation do not persist in traditional in vitro models due to selection pressures, thus requires exogenous overexpression. Alternatively, EGFR statuses can be maintained in xenografted mice, but implantation of the primary GBM cells into the flank is required to first establish the tumor prior to secondary injection into the brains. Recently, we have established a novel protocol for culturing GBM tissue as organoids (GBOs) directly from patient tumor resection that retain many distinct cell populations in vitro with high fidelity evidenced by histological, whole-exome, bulk and single cell RNA analyses. Compared to prolonged generation time of previously established in vitro and xenograft models, our methodology is robust for generating GBOs within 1–2 weeks from initial resection. In addition, these GBOs can be readily xenografted into the adult mouse brains as an intact organoid, exhibit rapid and aggressive infiltration phenotypes, and maintains driver mutation EGFRviii within as little as one month. Consequently, they can be used to test in vivo treatment efficacies in a timely fashion. The presence of diverse cell types in this GBO model offers a promising platform for not only understanding of tumor biology, but also more strategic development of new therapies.
TMOD-31. ADAPTING ENGINEERED CELL THERAPIES TO UNDERSTAND AND OVERCOME GLIOBLASTOMA RESISTANCE USING INTEGRATED IN VIVO AND EX VIVO MODELS
Authors: Satterlee Andrew , Dunn Denise , Floyd Scott , Hingtgen Shawn 
Published: November 2019

Abstract
Genetically engineered neural stem cells (NSCs) are a promising therapy for the highly aggressive brain cancer glioblastoma (GBM), yet treatment durability remains a major challenge. We sought to define the events that contribute to dynamic adaption of GBM during NSC treatment and develop strategies to convert initial tumor kill into sustained GBM suppression. Using a unique hybrid tumor model treated with human skin-derived induced NSCs (iNSCs) releasing the pro-apoptotic agent TRAIL, we investigated how spatial distribution of tumor and iNSCs affects GBM adaption throughout recurrence. Serial bioluminescent imaging (BLI) was used to track tumor volumes in vivo, while a subset of mice were sacrificed 6, 13, and 20 days post-treatment to harvest brains and generate living ex vivo tissue slices. Live animal imaging showed iNSC-TRAIL treatment rapidly decreased tumor volumes when delivered into the primary tumor mass; however, minimal impact on tumor growth was observed when cells were delivered into distal regions of the brain. In contrast, high-resolution imaging of living brain sections showed extensive impacts of iNSC-TRAIL therapy that could not be visualized with BLI. The living slices showed iNSC-TRAIL treatment into the primary tumor decreased the solid, but not the invasive, tumor burden. Treatment into the lateral ventricles did impact tumor kill and was more effective at treating the invasive tumor burden and maintaining inhibition than treatment into the contralateral parenchyma. We next utilized the living tissue slices to explore the sensitivity of the recurrent tumors to TRAIL. When therapy was applied to slices harboring recurrent tumor, treatment again significantly reduced tumor volumes, suggesting that tumors had not acquired TRAIL resistance. These results informed an additional in vivo survival study and subsequent PCR analysis of untreated and recurrent tumors, and combine the fidelity of in vivo studies with the speed and spatial resolution of living brain slice technology.
TMOD-32. GL261 LUCIFERASE-EXPRESSING CELLS ELICIT AN ANTI-TUMOR IMMUNE RESPONSE: AN EVALUATION OF MURINE GLIOMA MODELS
Authors: Sanchez Victoria , Lynes John , Walbridge Stuart , Wang Xiang , Edwards Nancy , Nwankwo Anthony , Dominah Gifty , Obungu Arnold , Adamstein Nicholas , Dagur Pradeep , Wu Tianxia , Maric Dragan , Munasinghe Jeeva , Heiss John , K Nduom Edjah 
Published: November 2019

Abstract
Preclinical models that reliably recapitulate the immunosuppressive properties of human gliomas are essential to assess immune-based therapies. Intracranially injected GL261 cells are widely used as an immunocompetent animal model of glioma, but it is common practice to transfect these with luciferase to facilitate tumor monitoring during treatment. Our group has previously shown that the luciferase-expressing GL261 Red-FLuc cells create an inflammatory response when implanted intracranially. Now, we additionally explore the inflammatory response of GL261-Luc2 cells and demonstrate a similar host immune response occurs with this model as well. In our in vivo evaluation, C57BL/6 mice underwent stereotaxic, intracranial implantation with GL261, GL261 Red-FLuc or GL261-Luc2 cells at doses of 5x104cells/5mL or 3x105cells/5uL.MRIs were performed to monitor relative tumor growth. To assess intrinsic differences between cell lines, in vitro cytokine profiles were evaluated by proteome microarray. Kaplan-Meier survival analyses demonstrated median survival for mice implanted with GL261 cells at 5x104cells was 18 to 21 days. The GL261-Red FLuc implanted mice cells did not reach median survival at either tumor dose with greater than 60% of mice termed long-term survivors. Finally, mice injected with GL261-Luc2 cells at 3x105cells reached median survival at 23 days, but median survival was significantly prolonged for mice implanted with GL261-Luc2 at a dose of 5x104cells (37 days, with 40% becoming long-term survivors) compared to GL261 implanted mice. MRIs reveal differences in tumor growth that correspond with the differences in median survival between groups. In addition, proteomic analyses revealed significantly elevated inflammatory cytokines such as IFN-gamma, IL-7 and TNF-alpha in the supernatants of the GL261 Red-FLuc cells and GL261-Luc2 cells. Further immune characterization is ongoing. Our data suggests that GL261 Red-FLuc and GL261-Luc2 murine models elicit an anti-tumor immune response by increasing pro-inflammatory modulators which stimulate the tumoricidal function of immune cells in the tumor microenvironment.
SCIDOT-25. ENHANCING THE EFFICACY OF DRUG RELEASED FROM LOADED FIBRIN GLUE BY PHOTOCHEMICAL INTERNALIZATION (PCI)
Authors: Hirschberg Henry , Nguven Lina , Johnsen Julie 
Published: November 2019

Abstract
Although gross total resection of gliomas is often possible with current available techniques, almost all tumors recur due to the infiltrative nature of these tumors. Infiltrative tumor cells in normal brain are consequently protected by the blood brain barrier (BBB), greatly reducing the efficacy of chemotherapy. Localized per-operative methods of slow-release drug delivery, that bypass the BBB altogether, are therefore of considerable interest. Fibrin glue (FG) is a FDA approved compound often used during surgery. FG loaded with drugs has been studied as a localized controlled release vehicle but the quantity and release kinetics attainable for some drugs using FG are often inadequate. Photochemical internalization (PCI) has been demonstrated to significantly enhance the efficacy of several chemo-theraputic agents such as bleomycin (BLM) and doxorubicin (DOX). In particular PCI of BLM has been shown to be effective in sterilizing the tumor bed after marginal cytoreductive surgery. We have evaluated the ability of PCI to enhance the ability of BLM and DOX, released from loaded FG, to inhibit the growth of multi-cell glioma spheroids in vitro. Spheroid growth was monitored for 14 days. The results of the present study show that drug was released for up to 72 hours. The growth inhibition caused by either BLM or DOX was significantly enhanced by PCI. The photosensitizer AlPcS2a could also be loaded and released from FG allowing local delivery at the tumor site.
SCIDOT-38. DEVELOPMENT OF AN IMAGE-INFORMED MATHEMATICAL MODEL OF CONVECTION-ENHANCED DELIVERY OF NANOLIPOSOMES FOR INDIVIDUAL PATIENTS
Authors: Woodall Ryan , Hormuth David , Abdelmalik Michael , Wu Chengyue , Feng Xinzeng , Phillips William , Bao Ande , Hughes Thomas , Brenner Andrew , Yankeelov Thomas 
Published: November 2019

Abstract
186-Rhenium nanoliposomes (RNL) are an experimental theranostic being investigated for the treatment of recurrent Glioblastoma. While traditional external beam therapy exposures healthy tissue to radiation, RNL has the potential to deliver extremely large doses (> 2000 Gy) of localized radiation, minimally exposing surrounding tissue. RNL is delivered directly to the malignancy by convection-enhanced delivery (CED) via intracranial catheter. For this reason, accurate and precise delivery of RNL to the target region is an imperative. While models of CED for molecular agents exist, we know of no such models for CED of liposomal nanoparticles. To that end, we are developing a patient-specific advection-diffusion model of RNL delivery and distribution, informed by pre-delivery quantitative magnetic resonance imaging (MRI) parameters, and validated by intra-delivery single-photon emission computed tomography (SPECT). Apparent liposome diffusivity and interstitial hydraulic conductivity are spatially informed by diffusion weighted MRI, while the clearance of interstitial fluid is spatially informed by the T1 enhancement ratio after contrast agent delivery. The model output is compared to SPECT images at two time points, acquired mid-way through and immediately following the RNL infusion. At the time of submission, the model has been calibrated by patient-specific data to match the spatiotemporal distribution of RNL in four patients. After calibration, the concordance correlation coefficient between the model and SPECT measurements was 0.80 +/- 0.23 mid-way through the infusion volume, and 0.86 +/- 0.14 immediately post-infusion. The DICE coefficient between the modeled delivery volume and measured delivery volume was 0.86 +/- 0.10 mid-way through the infusion volume, and 0.81 +/- 0.14 immediately post-infusion (reported as mean +/- 95% confidence intervals). These results provide preliminary evidence that the model can capture the spatiotemporal distribution of RNL during and after delivery, and may enable physicians to better plan CED procedures for liposomal nanotherapeutics in the future.