bluuebunny/crossref_metadata_2025_split huggingface dataset:
The mutational landscape of older patients with IDH wild-type glioblastoma (GBM).
Authors: Johnson Margaret O , Xiu Joanne , Glantz Michael J. , Zeng Jia , Chen Clark C , Dunbar Erin M. , Fonkem Ekokobe , Kesari Santosh , Brenner Andrew J. , Newton Herbert B. , Low Justin , Sumrall Ashley Love , Korn Wolfgang Michael , Ashley David M. 
Published: June 2021

e14033 Background: Advanced age is associated with poorer outcomes in GBM and current NCCN guidelines distinguish older GBM patients (≥70 years, oGBM) from their younger counterparts ( < 70 years, yGBM). We aim to characterize age-related comprehensive mutational profiles with the long-term goal of improving treatment strategies, outcomes, and rational clinical trial design. Specifically, we focused on IDH-wildtype (WT) oGBM, investigated if oGBM are more likely to acquire temozolomide-induced hypermutations, and finally, compared the frequency of high-tumor mutational burden (TMB) between oGBM vs. yGBM. Methods: Comprehensive molecular profiles of 1,657 adult IDH-WT GBM tumors tested at Caris Life Sciences (Phoenix, AZ) were queried. Tests included NGS of DNA (NextSeq, 592 Genes and NovaSEQ, WES) and RNA (NovaSeq) sequencing. SBS11 gene signature (i.e temozolomide-induced hypermutational profile) was queried using SigProfiler (Alexandrov 2020, Nature). Significance was determined by X2 and Fisher-Exact and p adjusted for multiple comparisons ( q) was < 0.05. Results: We identified 1,657 patients (range 21-89 years old, median 61 years) with IDH-wildtype GBM, 22% (360) of whom were ≥ 70 years. There was a slight male predominance (60%) for all ages. The most prevalent alterations in oGBM were TERT promoter mutation (105/131,80%), MGMT promoter methylation (pMe) (175/346, 51%), and PTEN mutation (129/349, 37%). EGFR amplification was seen in 35% (125/356) and EGFRvIII in 23% (81/360); Overall, fusions were seen in 12% (44 of 360) oGBM; events > 1% included MET (3.6%), FGFR3 (3.1%), EGFR (2.6%) and ROS1 (1.4%). 17% (56/349) of oGBM had positive PD-L1 by IHC. High TMB ( > 10mt/Mb) tumors were rare (3.1%) and MSI-high tumors even rarer (0.8%) in oGBM. When compared to yGBM, MGMT pMe was more prevalent (51% vs 38%, risk ratio (RR) 1.35 [1.19-1.52], q < 0.05) and NF1 mutations were less frequent in oGBM (21% v 34%, RR 0.62 [0.50-0.77], q < 0.05). No significant differences were seen in other key markers examined. The prevalence of SBS11 gene signature across all ages (data available for 1,141 patients) was 1.2% and was comparable across the age spectrum; no significant difference seen in the MGMT pMe group when oGBM was compared to yGBM (3.9% vs. 1.8 %, p= 0.3). Conclusions: This study represents the largest comprehensive molecular characterization of older IDH-WT GBM patients. We show that molecular profiles of IDH-WT GBM are remarkably similar across the age spectrum, including immunotherapy-associated markers, gene fusion landscape, EGFR amplification, and TMB. The significantly higher prevalence of MGMT pMe and lower NF-1 mutation rate in the older population bear significant prognostic and therapeutic implications.
BMP4 to increase connexin43 expression and interaction with β-catenin in glioblastoma stem cells.
Authors: Sutherland Natalia , Deaver Stacie , Wynne Taylor , Wheeler Christina , Lamouille Samy 
Published: June 2021

e14045 Background: Glioblastoma multiforme (GBM) is the most commonly occurring and most lethal primary central nervous system tumor. Treatment for GBM involves an aggressive combination of surgical resection, radiotherapy, and temozolomide chemotherapy. While intensive therapy has shown marginal improvement in prognosis, the median survival for GBM patients remains between twelve to fourteen months. The aggressiveness of GBM is attributed to the cellular heterogeneity and the invasive nature of the tumor, as well as the presence of glioblastoma cancer stem cells (GSCs). GSCs are a subpopulation of slowly dividing cells within GBM tumors that can survive chemotherapy and irradiation treatments targeting proliferating cells. The remaining quiescent GSCs can then enter an active state of self-renewal and differentiation through asymmetric division that contributes to tumor recurrence by recapitulating the heterogeneous tumor in relatively short order. Previously, we have identified a non-junctional, tumorigenic function for the gap junction protein connexin43 (Cx43) through its interaction with microtubules in GSCs. However, when GSCs are differentiated, this interaction decreases while Cx43 expression increases, suggesting an alternate function for Cx43. Bone Morphogenic Proteins (BMPs) have been implicated in GSC differentiation and their role in human tumorigenesis has remained largely unknown in GSCs. In this study, we aim to determine the effect of BMP on Cx43 expression and localization in GSCs. Methods: In this study, we used GBM patient-derived GSCs isolated from two GBM tumors following resection at Carilion Clinic, and maintained in stem cell medium. The effect of BMP4 signaling on Cx43 expression and localization in GSCs was determined using RT-qPCR, immunofluorescence, western blot, and co-immunoprecipitation. Results: Preliminary data show that BMP4 treatment increases Cx43 expression at the protein and mRNA levels through nuclear translocation and transcriptional activity of β-catenin. In addition, BMP4 treatment increases the interaction between Cx43 and β-catenin at the plasma membrane. Further studies are underway to determine the implications of disrupting Cx43/β-catenin interaction in GSC differentiation. Conclusions: While additional experiments are needed, our findings may be adapted as a prognostic indicator to determine patients’ susceptibility to traditional GBM therapies, and aid in novel therapeutic development to eradicate GSCs in GBM and prevent tumor recurrence following treatment.
Anti-PD-L1 treatment to enhance the response of glioblastoma to radiation and produce long-term survival in mice.
Authors: Wang Fei , Zhao Nan , Lin Chi , Zhang Chi 
Published: June 2021

e14048 Background: Glioblastoma (GBM) is the most aggressive and most common primary central nervous system cancer in adults. Blocking the interaction between Programmed Cell Death Protein-1 (PD-1) and its ligand (PD-L1) has shown remarkable success in the treatment of several cancers. However, many challenges remain in improving the efficacy of using monoclonal antibodies (Ab) against the receptor PD-1 in GBM, mainly due to the “non-immunogenic” tumor characteristics of GBM. PD-L1 has been found to be overexpressed on the surface of human GBM tumor cells and tumor-associated macrophages (TAM). Radiotherapy (RT), as one of the standard therapy of GBM, could alter the tumor microenvironment and promote an antitumor immune response. We hypothesize that anti-PD-L1 therapy can enhance the RT effects and improve the outcome of treatment when combined. Methods: Using a preclinical orthotropic syngeneic CT-2A mouse GBM tumor model, we studied the efficacy of combined therapy with anti-PD-L1 and RT. Mice were stratified into four treatment groups: control, RT, anti-PD-L1 Ab, and anti-PD-L1 Ab plus RT. RT(8 Gy) was given one time simultaneously with the first dose of anti-PD-L1, followed by systemic anti-PD-L1 maintenance treatment to the mice. Overall survival and tumor growth were monitored. Immunohistochemistry on resected tumors during treatment was performed to characterize the immune response. Single-cell RNA sequencing (scRNA-seq) was also performed to further study the immunologic parameters in the mouse brain. Results: Our results showed that anti-PD-L1 Ab in combination with RT provided a remarkable antitumor immune response and improved overall survival, with 25.5, 34, and 30 days of median survival in control (no-treatment), RT, and anti-PD-L1 groups, respectively, and achieving long-term survival and complete tumor response in 80% of the mice in the anti-PD-L1+RT treatment group (median survival not reached) in GBM tumor-bearing mice. The combined therapy promoted the recruitment of tumor-infiltrating immune cells, reversed the hostile tumor immune environment with a higher M1/TAM ratio, CD8+ /CD4+ T cell ratio, and CD8+ T cell /Treg cell ratio in the tumor area comparing with those parameters in single modality treatment groups. Furthermore, scRNA-seq data demonstrated that anti-PD-L1 combined with RT resulted in robust higher CD8 effector T cells, while lower CD4 and CD8 exhausted T cells in the tumor region compared to other groups. Increased CD4 central memory T cells and CD8 central memory T cells were seen only in tumors treated with anti-PD-L1+RT providing immunologic explanations on the durable control of GBM achieved only by the combined therapy. Conclusions: The anti-PD-L1 therapy synergizes with RT by reversing the hostile tumor immune environment resulting in improved tumor control and long-term survival in the syngeneic mouse GBM model.
Postoperative concurrent chemoradiotherapy plus apatinib in patients with malignant glioma: From one single research institute.
Authors: Fu Zhichao 
Published: June 2021

e14049 Background: Antiangiogenic therapy is a choice for the treatment of malignant glioma patients. Vascular endothelial growth factor receptors (VEGFRs) inhibitors block the new formation around the tumor and cut down the supply of oxygen, nutrients and metabolic waste, so that the tumor is hard to proliferate and metastases.Objective To explore the effectiveness and safety of apatinib in patients with malignant glioma. Methods: This is a retrospective and prospective case-control study conducted in a single center. Patients with new postoperative pathological diagnosis of malignant glioma (WHO IIĨIV) were selected. Enrolled patients received concurrent chemoradiotherapy (radiotherapy60Gy/30f/6w; TMZ75mg/m2,po,d1-d42) combined with or without apatinib (250mg, po, d1-d42,qd), then maintain 6 cycles(28 days a cycle)of TMZ chemotherapy(200mg/m2, d1-d5) .The primary endpoints were investigator-assessed progression free survival (PFS) and the grade of peritumoral brain edema (PTBE) evaluated by edema index(EI). The secondary endpoint was investigator-observed overall survival. Hazard ratios of PFS and OS were compared between trials in a Cox proportional hazards model. Results: Between October 2017 and March 2019, 47 patients (23 in apatinib group and 24 in control group) were enrolled in this study.The median age was 47 years in apatinib group (range from 19 to 73 years) versus 48 years in control group(range from 24 to 68 years). The results elucidated that the median PFS of the apatinib group was longer than that of the control group, but the difference was not statistically significant.(9.26 months vs. 6.69 months; hazard ratio, 0.65; 95% confidence interval [CI], 0.31 to 1.25; P=0.1906).As for median overall survival, the results of two groups were almost similar.(14.42 months vs. 14.36 months; hazard ratio, 0.74; 95%CI, 0.36 to 1.52; P=0.4036). Cox multivariate regression model revealed that apatinib was not a prognostic factor for PFS and OS (P>0.05). As of the statistical date, there were 7 patients with stable disease (SD), 16 patients with progressive disease (PD) and 13 patients with death in the apatinib group; whereas in the control group, the condition of 3 patients remained stable, the other 21 patients had progressed, with 19 unfortunate deaths. The grade of PTBE was improved in 15 of 23 patients (65.2%, 5/23 patients used corticosteroids) in apatinib group versus 6 of 24 (25%, 14/24 patients used corticosteroids) in control group. There was no grade 3 or 4 adverse events and serious adverse events. Conclusions: Compared with control group, apatinib group still improved mPFS by 2.57 months in patients with malignant glioma, but there was no statistical significance (P>0.05).The results also indicated that apatinib conferred a significant beneficial effect on PTBE improvement. All occurred adverse reactions were moderate and controllable. Clinical trial information: NCT03567135.
Generation of a HuTCR mouse platform-derived MAGE-A1-directed high-affinity TCR with superior potency versus human-derived TCRs.
Authors: Gavvovidis Ioannis , Leisegang Matthias , Oduro Jenifer , Obenaus Matthias , Leo Eugen , Blankenstein Thomas , Kieback Elisa 
Published: June 2021

e14515 Background: As cancer-testis antigens are self-antigens, T cells expressing high-affinity TCRs against such antigens are suppressed via negative thymic selection. Therefore, patient- or donor-derived TCRs are typically of low affinity and result in a reduced antitumor effect. Using our proprietary HuTCR platform, which consists of mouse lines carrying the full human TCR α/β loci in combination with common human HLA alleles, we have isolated high-affinity TCRs specific for the cancer-testis antigen MAGE-A1 and compared them to human-derived MAGE-A1-specific TCRs that are currently reported to be in clinical development. Furthermore, we validated MAGE-A1 as a potential cancer therapy target by using immunohistochemistry to evaluate expression in several major tumor types and healthy tissue. Methods: Using scRNAseq, TCRs were isolated from HuTCR mice. Human-derived MAGE-A1-specific TCR sequences were obtained from publicly available databases. All TCRs were expressed in primary human T cells as verified using peptide-MHC-multimer staining. Functional avidity of the TCRs was analyzed by coculture with T2 target cells loaded with titrated amounts of epitope peptides and measuring cytokine concentration by ELISA. Reactivity of TCRs to endogenously processed MAGE-A1 protein was assessed by co-culture with a panel of tumor cell lines varying in MAGE-A1 and/or MHC-class-I expression. MAGE-A1 expression on protein level was evaluated by immunohistochemistry. Results: Immunization of HuTCR mice with the antigen resulted in robust CD8+ T cell responses and several TCR clonotypes were identified by scRNAseq, with the majority of clonotypes being specific to the MAGE-A1-derived peptide KVLEYVIKV and TCR affinities ranging from 0.3 nM to 3 nM. By comparison, human-derived TCRs exhibited generally lower functional avidity from 3 nM to 60 nM. In addition, HuTCR-mouse-derived TCRs were more sensitive in recognition of tumor cell lines expressing low MAGE-A1 and/or HLA-A2. Immunohistochemical analysis of MAGE-A1 expression in healthy tissues demonstrated highly selective expression of MAGE-A1 in testis, only. Screening for expression confirmed that a significant proportion of several major cancer types expresses MAGE-A1 as reported by various other groups [reviewed in Curr Opin Cell Biol. 2015 December; 37: 1–8]. Conclusions: The HuTCR mouse platform allows for the generation of high-affinity MAGE-A1-specific TCRs with increased anti-tumor efficacy as compared to human-derived TCRs against the same cancer antigen. In addition, it was confirmed that MAGE-A1 has a highly selective expression pattern in healthy tissues (testis, only), but shows distinct expression in several major human tumor types.
The prevalence of HLA LOH in Chinese pan-cancer patients and its correlation with genomic alterations.
Authors: Zhao Jian , Xiao Xiaoxiong , Li Yue , Gao Xuan , Zhang Xiuqin , Liu Zeyi , Yi Yuting , Fu Xiaorui , Wang Han , Guan Yan-Fang , Xia Xuefeng , Zhang Weixing , Huang jian'An 
Published: June 2021

e14521 Background: Human leukocyte antigen (HLA) molecules are essential components involved in tumor antigen presentation. Loss of heterozygosity in HLA (HLA LOH) may facilitate tumor immune evasion. However, the large population study of HLALOH in Chinese pan-cancer patients remains to be explored. Methods: In this study, analysis were performed in 1504 advanced patients across 12 cancer types and 134 early-stage NSCLC patientsusing a 1021-gene panel. HLA LOH were analysis using LOHHLA algorithm. The consistency between target sequencing by 1021-gene panel and whole-exome sequencing (WES) was evaluated in 45 samples. Results: In 45 samples processed with both approaches, 95.6% (43/45) obtained concordant results. Among the 1638 patients, 24 were excluded due to homozygosity at all three HLA-Iloci. The prevalence of HLALOH in advanced patients presented considerable differences among cancer typeswith an average incidence of 45.6% (676/1482), ranging from 12.2% (prostate adenocarcinoma) to 68.0% (cervical squamous cell carcinoma). In NSCLC patients, there was an enrichment of HLALOH in both early-stage (I-IIIa) LUSC (24/88 [27.3%] of LUAD vs. 26/42 [61.9%] of LUSC, p<0.001) and advanced (IIIb-IV) LUSC. In 43.3% (293/676) patients harboring HLALOH, LOH co-occurred in HLA-A, B, and C. Consistent with previous studies, the occurrence of HLALOH was relevant to an elevated TMB level(median TMB 6.72 vs. 5.76, p<0.0001). In addition, the TMB level of patients with LOH at all three HLA-I loci was higher than those with LOH at only one HLA-I locus (median TMB 7.68 vs. 6.72, p=0.027).However, despite that MSI-H patients presented the highest incidence of TMB-H (TMB≥10.56), the incidence of HLALOH decreased in these patients. Alterations of multiple signaling pathways are enriched in HLALOH patients, including CPF (checkpoint factor) pathway, FA (Fanconi anemia) pathway, p53 pathway, RTK/RAS pathway, Notch pathway, Hippo pathway and Nrf2 pathway(p<0.0001, p=0.023, p<0.0001, p<0.0001, p=0.032, p=0.013, p=0.003, respectively, Table).Several oncogenes, such as TP53 and LRP1B, had higher alteration frequencies in the HLALOH group(p<0.001 for TP53,p<0.05 for LRP1B, Table 1). Conclusions: The 1021-gene panel could be applied for HLALOH analysis, provided that the relevant regions are well captured. The prevalence of HLALOH in Chinese cancer patients presents considerable differences across cancer types. Besides, the occurrence of HLA LOH was accompanied by higher alteration frequencies in several oncogenes and oncogenic signaling pathways. These insights may provide valuable information for clinical practice and follow-up research.[Table: see text]
Immune checkpoint inhibitor induced hepatitis: Risk factors, outcomes, and impact on survival.
Authors: Miah Abdul , Zhao Songzhu , Patel Sandip H. , Johns Andrew , Grogan Madison , Li Mingjia , Lopez Gabrielle , Haddad Tyler , Husain Marium , Kendra Kari Lynn , Otterson Gregory Alan , Wei Lai , Presley Carolyn J , Owen Dwight Hall 
Published: June 2021

e14525 Background: Immune checkpoint inhibitors (ICIs) have improved the survival of patients with multiple cancer types, however ICI treatment is associated with a unique set of immune-related adverse events (irAEs) that can affect any organ. Few studies have evaluated the risk factors and outcomes of ICI induced hepatitis (ICIH). Methods: We utilized an institutional database of patients with advanced cancers treated with ICI between 2011 and 2017 at The OSU Comprehensive Cancer Center to identify patients with ICIH. Any patient who received at least one dose of ICI alone or in combination with other systemic therapies either as part of clinical trial or standard of care were included. Clinical data were extracted through chart abstraction. irAEs were graded using the Common Terminology Criteria for Adverse Events v5. Overall survival (OS) was calculated from the date of ICI initiation to death from any cause or the date of the last follow-up. OS with 95% confidence intervals were estimated using the Kaplan–Meier method. OS was also evaluated by occurrence of ICIH using the log-rank test. Results: We identified 1,096 patients treated with at least one dose of ICI. Most common cancers included lung (n=224, 20%) and melanoma (n=342, 31%). The most common ICIs were PD1/L1 (n=774, 71%) and CTLA-4 inhibitors (n=195, 18%). ICIH of any grade occurred in 64 (6%) patients. Overall, 46 (71%) were male and median age was 60 years. Severity of hepatitis was grade 1-2 in 30 patients (47%) (Table 1). The incidence of ≥grade 3 ICIH in the entire cohort was 3.1%. Median time to ICIH diagnosis was 63 days. ICIH occurred alone in 24 patients, and co-occurred with other irAEs in 40 patients. The most common co-occurring irAEs were pneumonitis (n=7); colitis (n=15), thyroid abnormality (n=14); and dermatitis (n=15). ICIH was more common in women (p=0.038), in patients treated with combination ICIs (p<0.001), and among patients receiving first line therapy (p=0.018). Patients who developed ICIH had significantly longer OS than patients who did not develop ICIH; there was no difference in OS between patients with ≥grade 3 ICIH vs grade 1-2 (Table). 33 out of 34 patients with ≥grade3 ICIH were treated with steroids; 3 received mycophenolate and one received infliximab. Of patients with ≥grade 3 ICIH, 11 resumed ICI therapy without recurrent ICIH. Conclusions: Female sex, combination immunotherapy, and line of therapy were associated with ICIH. Patients with ICIH had improved clinical outcomes compared to those that did not develop ICIH, even those with higher grade toxicity. Further study is needed to assist in developing risk stratification models and optimal treatment for ICIH. OS of patients with and without immune checkpoint inhibitor hepatitis.[Table: see text]
Modelling treatment benefit for bexmarilimab (an anti-Clever-1 antibody and a novel macrophage checkpoint inhibitor) using phase I first-in-man trial data.
Authors: Bono Petri , Ekström Jussi , Karvonen Matti K , Mandelin Jami , Koivunen Jussi 
Published: June 2021

e14530 Background: Bexmarilimab, an investigational immunotherapeutic antibody targeting Clever-1, is currently investigated in phase I/II MATINS study (NCT03733990) for advanced solid tumors. Machine learning (ML) based models combining extensive data could be generated to predict treatment responses to this first-in-class macrophage checkpoint inhibitor. Methods: 58 baseline features from 30 patients included in the part 1 of phase I/II MATINS trial were included in ML modelling. Seven patients were classified as benefitting from the therapy by RECIST 1.1 (PR or SD response in target or non-target lesions). Initial feature selection was done using a combination of domain knowledge and removal of features with several missing values resulting in 20 clinically relevant features from 25 patients. The remaining data was standardized and feature selection using variance analysis (ANOVA) based on F-values between response and features was performed. With this approach, the number of features could be further reduced as the prediction performance increased until the most important features were included in the model. Several prediction models were trained, and prediction performance evaluated using leave-one-out cross-validation (LOOCV), with and without SMOTE oversampling of the positive class of the training data inside each LOOCV fold. In LOOCV the prediction model was trained 25 times. Stacked meta classifier with SMOTE oversampling combining three classifiers: elastic-net logistic regression, random forest and extreme gradient boosting was chosen as the best performing prediction model. Results: Seven baseline features were associated with bexmarilimab treatment benefit. Increasing bexmarilimab dose and high tumor FoxP3 cells showed positive benefit. On contrary, high baseline blood neutrophils, CD4, T-cells, B-cells, and CXCL10 indicated negative relationship to the treatment benefit. The ML model trained with these seven features performed well in LOOCV as 6/7 benefitting and 16/18 non-benefitting were classified correctly, and all considered classification performance metrics were good. In feature importance analysis, low baseline CXCL10 and neutrophils were characterized as the most important predictors for treatment benefit with values of 0.19 and 0.16. Conclusions: This study highlights possibility of using ML models in predicting treatment benefit for novel cancer drugs such as bexmarilimab and boost the clinical development. These findings are in line of expected immune activation of bexmarilimab treatment. The generated ML models should be further validated in a larger patient cohort. Clinical trial information: NCT03733990.