bluuebunny/crossref_metadata_2025_split huggingface dataset:
TMET-08. METABOLOMIC CHANGES IN GLIOBLASTOMA PATIENTS UNDERGOING CONCURRENT CHEMORADIATION THERAPY
Authors: Aboud Orwa , Liu Yin Allison , Fiehn Oliver , Bridges Christopher , Fragoso Ruben , Hodeify Rawad , Bloch Orin 
Published: November 2022

Abstract

OBJECTIVE
To illustrate changes of untargeted metabolomics in patients with glioblastoma IDH wildtype undergoing concurrent radiation therapy (RT) with temozolomide (TMZ). This study implemented machine learning (ML) algorithms to predict treatment phase: pre-surgery, post-surgery, pre-radiation, and post radiation based on untargeted metabolomics data.

METHODS
Thirty-six patients with glioblastoma IDH wildtype (18 methylguanine methyltransferase [MGMT] methylated, 16 MGMT unmethylated, 2 MGMT status unknown) were enrolled into this study. Serum samples obtained from patients on the same day before surgery, 2 days after surgery, before starting their concurrent chemoradiation, and after concluding this phase of treatment. Blood samples were obtained via antecubital phlebotomy without regard for time of the day, diet, or fasting status. Untargeted metabolomics by GC-TOF mass spectrometry were obtained and compared. The proposed ML models analyzed 105 samples from 36 patients utilizing 157 structurally identified blood metabolites. Multinomial Logistic Regression (MLR) and GradientBoostingClassifier (GB Classifier) were used to classify patient samples based on detected changes in blood metabolites. The classification performance of these models was evaluated using performance metrics and AUC scores.

RESULTS
Post radiation; significant increase in the following metabolites: glycine, serine, threonine, oxoproline, 6-deoxyglucose, gluconic acid, glycerol-alpha-phosphate, ethanolamine, propyleneglycol, triethanolamine, xylitol, and fumaric acid were noted while significant decrease in 3-aminopiperidine 2,6-dione was noted post radiation. MLR produced 78% accuracy, 75% precision, and AUC = 0.89, and GB Classifier produced 75% accuracy, 77% precision and AUC = 0.91. Finally, we presented a pattern of metabolites changes per clinical stage based on pairwise correlations.

CONCLUSIONS
This study represent the first serum metabolic signature associated with RT in patients with glioblastoma. The results from the classification algorithms and pairwise correlations showed that blood metabolites have the potential to predict phase of treatment and potentially enable to evaluate response to treatment in patients with glioblastoma in a relatively small cohort.
TMET-10. FATTY ACID METABOLISM REGULATES IMMUNE REACTIVITY OF IRRADIATED GLIOBLASTOMA
Authors: De Martino Mara , Daviaud Camille , Laiakis Evagelia , Vanpouille-Box Claire 
Published: November 2022

Abstract
Radiation therapy (RT) is the standard-of-care for glioblastoma (GBM) and is the only treatment for inoperable brain tumors. In multiple cancer, RT stimulates anti-tumor immunity by at least activating cancer cell-intrinsic type I interferon (IFN-I) responses. However, GBM inevitably recur, suggesting that RT promotes immune evasion mechanisms. Altered fatty acid (FA) metabolism is an emerging mechanism that can account for treatment resistance and immune escape of GBM. Therefore, we hypothesize that RT induces a metabolic shift toward the production of FAs to foster immunosuppression of GBM. Supporting this hypothesis, our data show that RT upregulates the expression of the fatty acid synthase (FASN) and the activity of the fatty acid desaturase 2 (FADS2) to generate FAs in murine GBM models, GL261 and CT2A. To determine whether FA metabolism was preventing RT-induced IFN-I, we impaired FA synthesis by inhibiting either FASN or FADS2 using genetic (CRISPR-Cas9) or pharmacological inhibitors (FASNi or FADS2i). As expected, protein quantification (ELISA) as well as gene expression (RT-qPCR) revealed that irradiated GBM cells released greater levels of IFN-beta 24hrs post RT; an effect that was more pronounced when FASN or FADS2 were blocked. Reinforcing the role of FA metabolism in regulating immune activation, in situ immunofluorescence of irradiated intracranial GL261 tumors showed that mice treated with FASNi improved the recruitment of CD8+ T cells into GBM. Along similar lines, flow cytometry of digested GL261 tumors revealed an increase infiltration of CD11c+ CD103+ dendritic cells as well as activated CD86+ CD11b+ cells when FAs synthesis was blocked with FADS2i. Altogether, our data suggest that RT-induced FA synthesis represents a mechanism of resistance by preventing IFN-I and promoting immunological evasion of GBM. Targeting FA metabolism is a promising strategy to promote anti-tumor immunity and sensitize irradiated GBM to immunotherapies.
TMET-13. SEX DIFFERENCES IN GLUCOSE METABOLISM AND MITOCHONDRIAL FUNCTION IN GLIOBLASTOMA IMPLICATE HYPOXIA-INDUCIBLE FACTOR 1 ALPHA (HIF1A) ACTIVITY
Authors: Lu Jianyun , Sponagel Jasmin , Jones Jill , Gass Hannah , Nunez Elena , Frankfater Cheryl , Williams-McLeod Sierra , Marquez Camryn , Chinnaiyan Prakash , Rubin Joshua , Ippolito Joseph 
Published: November 2022

Abstract
Glioblastoma (GBM) is the most common and aggressive primary brain tumor in adults. It is more prevalent in males and female patients have better survival. Investigating the molecular mechanisms underlying this disparity is imperative for understanding its development and progression as well as developing novel treatment paradigms. Carbohydrate (namely glucose) metabolism is a critical GBM nutrient source for biosynthesis, energetics, and reducing equivalents. Previously, our group discovered that elevated glycolytic activity uniquely predicted the outcomes of male, but not female, lower grade glioma patients. Our goal was to characterize sex differences in GBM carbohydrate metabolism and their effects on cellular phenotype. First, we discovered that male transformed murine astrocytes were more susceptible to glucose deprivation than females. We confirmed this phenotype with irreversible inhibition of hexokinase with 2-deoxyglucose as well as a GLUT1-selective inhibitor. Time-resolved stable isotope tracing of cell metabolism with carbon-13 glucose in transformed astrocytes further supported these findings; male cells had significantly higher rates of glucose uptake and metabolism than female cells. These results were validated with stable isotope metabolomics datasets from human cancer cell lines. Using additional assays of cellular metabolism, we discovered that male transformed astrocytes had a higher glycolytic rate, higher pyruvate kinase activity, higher mitochondrial respiration, and higher mitochondrial mass compared to females. This was validated by a TCGA pancancer analysis that revealed significantly higher expression of nuclear genes involved in mitochondrial regulation in males than in females. This prompted us to identify possible regulators of this metabolic phenotype. We discovered that HIF1A had robust hypoxia-inducible expression that was significantly higher in male transformed astrocytes. Moreover, HIF1A expression as well as its target transcripts were significantly higher in TCGA pancancer tumor datasets. Together, our data underscore the potential for developing sex-specific metabolic targeting approaches for patients with GBM.
TMET-16. DIRECT DETECTION OF 2HG AND GLUTAMATE PRODUCTION USING HYPERPOLARIZED [1-13C-5-12C]-Α-KETOGLUTARATE IN CELL AND IN VIVO GLIOMA MODELS
Authors: Hong Donghyun , Kim Yaewon , Mushti Chandrasekhar , Minami Noriaki , Gillepspie Anne Marie , Batsios Georgios , Taglang Celine , Cherukuri Murali , Wu Jing , Viswanath Pavithra , Swenson Rolf , Vigneron Daniel , Ronen Sabrina 
Published: November 2022

Abstract
Mutant IDH leads to 2HG production, which drives glioma development. 13C MRS monitoring of hyperpolarized [1-13C]α-ketoglutarate (αKG) metabolism to 2HG and glutamate provides a non-invasive assessment of the IDH mutation and normal metabolism, respectively. However, monitoring 2HG production in vivo has been challenging because its resonance is within 0.1 ppm of the natural abundance [5-13C]αKG signal of the [1-13C]αKG substrate. Here, we utilized [1-13C -5-12C]αKG, which eliminated the [5-13C]αKG peak. This new approach, combined with an optimized sequence, made it possible to readily monitor the production of both 2HG and glutamate in cells and in vivo in healthy rats or rats with orthotopic patient-derived glioma. Hyperpolarized [1-13C -5-12C]αKG was injected into genetically engineered NHAIDHmut cell lysates, healthy rats, and rats implanted orthotopically with BT257 cells intravenously. 1-D dynamic 13C MRS spectra from cells and in vivo slab spectroscopy data were then acquired using an 11.7 T NMR system and a 3 T pre-clinical scanner, respectively. Injection of the hyperpolarized [1-13C -5-12C]αKG into cell lysates showed clearly detectable dynamic conversion of hyperpolarized [1-13C-5-12C]αKG to 2HG and glutamate. The normal brain showed clear production of glutamate but no 2HG was detected. In tumor-bearing rats, we were able to clearly detect the dynamic production of both 2HG and glutamate. This study demonstrated the utility of hyperpolarized [1-13C-5-12C]αKG as a substrate to clearly assess 2HG production without the confounding presence of the natural abundance peak which cannot be distinguished from 2HG in vivo. Importantly, the detection of 2HG provides a clear indicator of the IDH mutation within the tumor.
TMET-21. ENHANCING THE METABOLIC FITNESS OF T CELLS TO TREAT BRAIN TUMORS
Authors: Ghosh Tanya , Jin Linchun , Feier Diana , Silver Aryeh , Tian Guimei , Dajac Mariana , Rahman Maryam , Flores Catherine , Sarkisian Matthew , Huang Jianping , Mitchell Duane , Deleyrolle Loic 
Published: November 2022

Abstract

INTRODUCTION
Glioblastoma is a challenge for neuro-oncologists and current therapies are minimally effective. Standard-of- care treatment is almost inevitably followed by disease recurrence. Adoptive T cell transfer has emerged as a viable therapeutic for brain malignancies. While promising, the efficacy of this approach is often limited by a complex immunosuppressive tumor microenvironment. These complexities mean that more sophisticated T cell products are required. Objectives: The brain tumor microenvironment provides local restraints via metabolic competition suppressing antitumor immunity, specifically inhibiting infiltration and tumoricidal functions of host and adoptively transferred tumor-reactive T cells. The overall goal of this project is to test new treatments to reverse immune dysfunction in cancer through the regulation of T cell metabolic signaling. We propose that modulating the glucose pathway in T cells can potentiate their anti-tumor activity once adoptively transferred.

METHODS
The glucose metabolic pathway of T cells was modulated via overexpression of glucose transporters. The functionality of metabolically modified T cells was investigated in murine and human models.

RESULTS
We demonstrated the existence of a competition for glucose between T cells and tumor cells, with tumor cells imposing glucose restriction mediating T cell hyporesponsiveness. Overexpression of glucose transporters such as Glut1 and Glut3 increased T cell glucose utilization and provided a survival/growth advantage and enhanced T cell activation in glucose-restricted conditions. We also established that glucose transporter overexpression improves intratumoral infiltration and expansion of adoptively transferred T cells, resulting in improved survival.

CONCLUSION
This project integrates fundamental concepts of tumor and immune metabolism in the design of immunotherapy and confirms that immunometabolism represents a viable target for new cancer therapy to treat brain tumors.
TMET-22. ENHANCED LIPID METABOLISM IN BRAIN TUMORS: AN EXPLOITABLE METABOLIC VULNERABILITY THROUGH DIET-INDUCED KETOSIS
Authors: Kant Shiva , Kesarwani Pravin , Zhao Yi , Chinnaiyan Prakash 
Published: November 2022

Abstract
Enhanced lipid metabolism contributes towards carcinogenesis at multiple levels, serving as a "gain of function" in several cancers, including glioblastoma. Seemingly paradoxical to these findings, pre-clinical investigations have demonstrated anti-tumor activity in mice fed a high fat/low carbohydrate ketogenic diet (KD), both alone and in combination with conventional therapy. Therefore, we sought to define mechanistic underpinnings contributing towards this apparent disconnect. Consistent with previous reports, mice fed a KD demonstrated independent anti-tumor activity and potent synergy with RT when tested in an aggressive glioblastoma orthotopic model. To provide a window into metabolic consequences of a KD in glioblastoma, we performed global metabolomic profiling on tumors and serum from mice fed a standard diet and KD. Although ketosis was confirmed, no change in glucose was observed in serum of KD mice, suggesting presence/absence of carbohydrates might not be contributing to the observed anti-tumor activity. Interestingly, profound intra-tumoral metabolic changes were observed in intracranial tumors from mice fed a KD, with an accumulation of unsaturated fatty acids emerging as a central metabolic node. Intriguingly, recapitulating these findings in culture conditions in vitro, polyunsaturated fatty acid linoleic acid demonstrated anti-proliferative activity, decreased clonogenic capacity, and synergy with RT in GBM cells, recapitulating in vivo findings. This was not observed when cultured with the monounsaturated fatty acid oleic acid. Through a series of investigations, we went on to identify lipid storage homeostasis and lipotoxicity associated with free fatty acid accumulation played a contributory role in the differential activity of these different classes of fatty acids. Importantly, PUFA-rich modified diet showed potent antitumor activity in combination with RT in our preliminary investigation. These studies collectively reaffirm and provide mechanistic underpinnings of the anti-tumor activity of a KD, suggesting that enhanced lipid metabolism in brain tumors may serve as an exploitable metabolic vulnerability through diet modification.
TMET-23. GLIOBLASTOMA EXHIBIT SEX DIFFERENCES IN BRANCHED-CHAIN AMINO ACID METABOLISM
Authors: Sponagel Jasmin , Gass Hannah , Cho Kevin , Chinnaiyan Prakash , Patti Gary , Rubin Joshua , Ippolito Joseph 
Published: November 2022

Abstract
Glioblastoma (GBM) is the most common and aggressive primary brain tumor in adults. It is more prevalent in males and female patients exhibit a survival advantage. Understanding the molecular mechanisms that underlie those sex differences could support novel treatment strategies. Amino acids, such as glutamine and the branched-chain amino acids (leucine, isoleucine, valine; BCAAs), play important roles in GBM as they are involved in biosynthesis, energetics, redox homeostasis, and mTOR signaling. Using murine transformed astrocytes, we recently discovered that male cells are more dependent on glutamine and utilize more glutamine to replenish biosynthetic pathways, including BCAA regeneration. BCAA regeneration (i.e., re-amination of branched-chain ketoacids to BCAA) is catalyzed by branched-chain aminotransferases (BCATs) and requires glutamate as a nitrogen donor. Importantly, BCAA regeneration has been linked to cancer progression and poor outcome. Using stable isotope glutamine labeling we found that male transformed astrocytes exhibit greater nitrogen transfer from glutamate onto branched-chain ketoacids to regenerate BCAAs. In line with increased glutamate utilization for BCAA regeneration via BCAT, we found that male transformed astrocytes exhibit greater BCAT1 protein expression levels. Concordantly, male transformed astrocytes are more sensitive to BCAA deprivation compared to female cells, indicating that BCAA metabolism may play a sex-specific role in GBM. Using human GBM surgical specimens, we found greater BCAA metabolite abundance in male GBM. Importantly, our findings on sex differences in BCAA abundance and dependency in GBM are in line with previous reports that show that BCAA abundance and utilization differ between males and females under normal physiological conditions. Together, these data suggest that male GBM are more dependent on BCAAs and that BCAT1 may play unique roles in male and female GBM metabolism. Our data highlight that GBM metabolism differs between males and females and underline the importance of considering sex in metabolic targeting approaches.
CNSC-23. NEURONAL ORIGIN INFLUENCES SPONTANEOUS NETWORK ACTIVITY IN GLIOBLASTOMA-NEURON COCULTURES
Authors: Daniel Andy , Krishna Saritha , Popova Galina , Kaur Jasleen , Hervey-Jumper Shawn L 
Published: November 2022

Abstract
Neuronal activity is emerging as a driver of cancer initiation and proliferation. Several models have been developed to recapitulate cancer-neuron interactions within the tumor microenvironment; however, there is substantial diversity of neuronal subtypes within the murine and human cortex. Differences in the neuronal microenvironment may influence glioma cell behavior and fail to recapitulate in vivo mechanisms. Here, we investigate the electrophysiological properties of cortical neurons and activity-regulated paracrine signaling across co-culture conditions using microelectrode arrays (MEA). Cortical neurons were obtained from three conditions, (1) human prenatal tissues, (2) murine embyronic tissue (E18), and (3) murine postnatal tissue (P1.5), then co-cultured with primary patient-derived glioblastoma cultures. Neuronal activity was assessed using weighted mean firing rate (WMFR; spike rate multiplied by number of active electrodes) and network burst synchrony index. Characterization of the activity-dependent paracrine signaling was performed using proteomics. Initiation of spontaneous firing activity differed across neuronal conditions, with activity occurring earlier in postnatal P1.5 cultures (day in vitro [DIV] 7), whereas E18 and human prenatal cultures demonstrated delayed patterns of activity (DIV 14 and DIV 21, respectively). Glioblastoma cells were added on the first day of consistent spiking. All neuronal co-culture conditions demonstrated glioma-induced hyper-activity (increased WMFR), however in different patterns. E18 co-cultures exhibited glioma-induced hyperexcitability within 24 hours with a subsequent decrease in activity over 14 days with minimal synchrony. P1.5 demonstrated no glioma-induced hyper-excitability at 24 hours; however, WMFR peaked by 7 days and remained elevated through co-culture day 14 (including concurrent increase in synchrony). Human postnatal neurons demonstrated a steady and consistent increase in WMFR as early as 24 hours post co-culture and remained active through day 14 with no synchrony change. Our data suggest that careful consideration should go into selecting neuronal co-culture conditions for mechanistic studies.
TMIC-03. TUMOR INFILTRATING LYMPHOCYTES IN BRAIN METASTASIS: CLINICOPATHOLOGICAL EVALUATION AND COMPARISON WITH PAIRED PRIMARY
Authors: Sahay Ayushi , Indave Mayur , Chatterjee Abhishek , Moiyadi Aliasgar , Patil Vijay , Gupta Tejpal , Shetty Prakash , Singh Vikas , Menon Nandini , Sahu Arpita , Chaudhari Amit , Sridhar Epari 
Published: November 2022

Abstract

INTRODUCTION
Little is known about tumour-infiltrating lymphocytes (TILs) in brain metastasis (BM) and their utility as predictive biomarker for immunotherapy. AIM: To analyze TILs in BM, and correlate with corresponding paired primary (PP).

METHODS
Inclusion: Histologically confirmed BM and PP (2015-2020). Exclusion: Hematological malignancies, spinal cord/meningeal metastasis, pediatric age, slides/blocks not available. Final Nf 277 BM; 64 PP. One H&E slide from both PP &BM assessed for stromal TILs (sTILs) and intratumoral TILs (iTILs) as per International TIL Working Group guidelines.

RESULTS
Median sTILs (10%) were significantly higher than iTILs (1%) in primary and BM (p-value &lt; 0.001). sTILs were mild (&lt;10%) in majority of primary (57.8%) and BM (59.9%), while iTILs were nearly always mild (primary 97.1%; BM 96.9%). Between PP and BM, no change in sTIL category in 48.43%, while conversion in 51.56%, with higher to lower conversion (66.67%) &gt; lower to higher (33.33%). Conversion rate higher for breast (54.3%) &GIT (62.5%) than lung (45.5%) &GUT (40%). In BM, preoperative steroid therapy (PST) showed lower sTILs (≤10%) (p-value= 0.041), and adenocarcinoma histology higher sTILs ( &gt;10%) (p-value= 0.001), while precocious metastasis (p-value= 0.051) and absence of extracranial metastasis (p-value= 0.083) trended towards higher sTILs. Median survival of whole cohort was 20 months (95% CI 14.4-25.68 months). Presence of extracranial metastasis (p-value= 0.01); precocious BM (p-value= 0.007) showed lower overall survival (OS), while adenocarcinoma histology better OS (p-value= 0.029), in both univariate and multivariate analysis. Absence of PST showed longer median survival, though not significant (24.1 vs 19.8months; p-value= 0.19).

CONCLUSIONS
sTILs predominantly show high &gt;low conversion between PP and BM, signifying lowered immune response in BM. Thus, evaluation of sTILs in BM, wherever tissue available, may be indicated if immunotherapy is considered. Parameters with higher sTILs showed longer survival, possibly indicating prognostic role of tumor microenvironment in BM.
TMIC-06. ANTAGONISM OF DRD2 USING ONC201 INCREASED EXPRESSION OF ANTIGEN PRESENTATION PATHWAY PROTEINS IN DIFFUSE MIDLINE GLIOMA, RECRUITING TUMOR INFILTRATING LYMPHOCYTES IN VIVO
Authors: Persson Mika , Jackson Evangeline , Duchatel Ryan , Bramberger Liesl , McEwen Holly , Kearney Padraic , Findlay Izac , Douglas Alicia , Kobbe Birgit , Wagener Raimund , Larsen Martin , Faridi Pouya , Holst Jeff , Mayall Jemma , Gedye Craig , Hondermarck Hubert , Horvat Jay , Nixon Brett , Cartaxo Rodrigo , Koschmann Carl , Valdes-Mora Fatima , Ortega David Gallego , Nazarian Javad , Alonso Marta M , Hulleman Esther , Van der Lugt Jasper , Vitanza Nicholas , Mueller Sabine , Dun Matthew 
Published: November 2022

Abstract
Diffuse midline glioma (DMG) is a high-grade glioma with a median overall survival of 9-11 months. Radiotherapy is the only recognized treatment. The DMG tumor microenvironment (TME) contains few, if any, tumor infiltrating lymphocytes (TILs) or inflammatory cytokines, thus is distinctive of an ‘immunologically cold’ tumor/TME.1 DMG lack the expression of immunosuppressive immune checkpoint proteins, likely explaining the failure of immune checkpoint inhibitors (ICI) tested under clinical trials for DMG patients, and suggestive of an alternative mechanism underpinning the cold TME. 1 Glioblastomas also harbor a cold TME, which can be somewhat explained by T cell lymphopenia, influenced by the sequestration of T cells in the bone marrow (through Beta-arrestin-induced internalization of Sphingosine-1-phosphate receptor 1 [S1PR1]). 2 Dopaminergic activation of Beta-arrestin and hence S1PR1 internalization, is potentially regulated through dopaminergic peripheral nerves in primary and secondary lymphoid organs, regulated by the Dopamine receptor D2 (DRD2), that is highly expressed on T cells. ONC201 is a potent DRD2 antagonist, currently in phase I-III clinical trials for DMG patients, alone and in combination with radiotherapy and the PI3K/AKT inhibitor paxalisib (NCT05009992). Proteomic profiling of DMG patient-derived cells +/-ONC201 showed increased expression of several antigen presenting pathway proteins, including Beta-2-microglobulin (B2M) and HLA class I histocompatibility antigen, A alpha chain (HLA-A). This was confirmed in vivo using SU-DIPG-VI patient-derived xenograft mouse model tissues +/-ONC201 alone, and together with paxalisib. Excitingly, this combination (given orally) promoted the recruitment of TILs to the tumor, revealing novel immunomodulatory effects. In vivo, ONC201 promoted the expression of EMILIN-3, a TGF-β antagonist that is known to inhibit HLA-A/B2M expression, possibly explaining the increased MHC-I activity. This study uncovers a novel link between treatment of DMG with ONC201 and paxalisib and the role dopaminergic peripheral nerves signaling may play on the sequestration of T cells within lymphoid organs and lymphopenia.