bluuebunny/crossref_metadata_2025 huggingface dataset:
Safety and efficacy of hypofractionated stereotactic radiosurgery with anlotinib targeted therapy for glioblastoma at first recurrence: A preliminary report.
Authors: Guan Yun , Zhu Huaguang , Li Jing , Mei Guanghai , Liu Xiaoxia , Li Chao , Gong Xiu , Dai Jiazhong , Pan Li , Wang Yang , Wang Enmin , Wang Xin
Published: June 2021

e14015

Background: The optimal treatment for recurrent glioblastoma (rGBM) remains uncertain. Hypofractionated stereotactic radiotherapy (HSRT) and anti-vascular endothelial growth factor (VEGF) antibodies (e.g, bevacizumab and anlotinib) have been reported a promising survival benefit and acceptable toxicity in recent studies. This research aimed to investigate the efficacy and safety of hypofractionated stereotactic radiosurgery with anlotinib as a salvage treatment for rGBM within the radiation field at first recurrence. Methods: Between December 2019 and June 2020, rGBM patients treated with CyberKnife (CK) radiosurgery concurrently with anlotinib were retrospectively analyzed. Anlotinib was prescribed 12 mg daily during CK. Adjuvant anlotinib was administered 12mg d1-14 every 3 weeks. The primary endpoint was objective response rate (ORR) using response assessment in neuro-oncology (RANO) criteria, and secondary endpoints included overall survival (OS), progression-free survival (PFS) after salvage treatment, and toxicity. Toxicity was assessed using common terminology criteria for adverse events (CTCAE) 5.0. Results: A total of five patients were included in the study. The median planning target volume (PTV) was 26.94 cm3 (5.53 to 54.41 cm3). The prescribed dose was 25.0 Gy in 5 fractions. The median number of cycles of anlotinib was 9 (4 to 15 cycles). The median baseline Karnofsky Performance Status (KPS) is 80 (70 to 90). The follow-up ranged from 4 to 10 months. Three (60%) patients had the best outcome of partial response (PR), and 2 (40%) achieved complete response (CR), the ORR was 100%. No patient died or had progressive disease (PD) until the last follow-up. Two patients had grade 2 hand-foot syndrome and relieved after dermatology treatments, no other grade 3 or higher toxicities was recorded. Conclusions: Salvage radiosurgery with anlotinib showed a favorable outcome and acceptable toxicity for rGBM patients in this preliminary report. A prospective phase II study (NCT04197492) is ongoing to further investigate the regimen in rHGG. [Table: see text]
Molecular characteristics of H3 K27M mutation gliomas in Chinese adults.
Authors: Zhang Peng , Han Tiantian , Lu Yi , Sun Weijie , Deng Wanglong , Lu Guanghua , Guo Didi , Li Xiaomin , Bu Fanfeng
Published: June 2021

e14018

Background: Diffuse midline glioma H3 K27M–mutant is a specific entity added to the 2016 update of the WHO classification of CNS tumors. H3K27M-mutant gliomas are diagnosed primarily in children and adolescents with TP53 and/or ATRX mutations. However, the characteristics of H3 K27M-mutant gliomas in adults have not been explicitly described. Methods: We performed the 131-gene panel targeted sequencing on tumor samples from 33 adults H3 K27M-mutant gliomas( > 18 years) and 13 children and adolescents H3 K27M-mutant gliomas(≤18 years) in a CAP certified laboratory. Somatic mutations, copy number variations, and fusion genes were detected following the standard operating procedure (SOP). The MS-based assay measured MGMT promoter methylation. We calculated the tumor location and the median age in different cohorts. Results: In our adult cohort, 22/33（66.7%）had a midline location（spinal cord n = 2, thalamus n = 7, brainstem n = 6, cerebellum n = 3, pineal region n = 1, Basal ganglia region n = 3）, 6/33（18.1%）had a non-midline location (Lateral ventricle n = 2, Cerebral hemispheres n = 4). In the children and adolescents cohort, 11/13（84.6%）occurred in midline location, 1/13(7.7%) occurred in the Lateral ventricle. MGMT promoter methylation did not differ in adult and pediatric H3 K27M-mutant gliomas (12.1% vs. 0). H3 K27M-mutant adult gliomas significantly co-occurred with the NF1 mutation(P = 0.008937). The median age of H3 K27M-mutant adult gliomas with NF1 modification (13/33, 39.4%) is higher than the NF1 wild type (49 years vs. 38 years, P = 0.107), although the difference has no statistical significance. Conclusions: In Chinese adults, as in children, H3 K27M mutation gliomas are characterized by a constant midline location, low rate of MGMT promoter methylation. Inconsistently, H3 K27M mutant adult gliomas are featured by a higher rate of NF1 mutations. Our molecular profiling analysis revealed the H3 K27M mutation in adult gliomas. Our research suggests potential molecular pathogenesis of H3K27M mutant adult gliomas and identifies more therapeutic targets for precision medicine.
Comparison of next-generation sequencing and FISH/IHC for molecular classification in glioma.
Authors: Jia Deze , Lu Yi , Li Xiaomin , Han Tiantian , Deng Wanglong , Bu Fanfeng , Lu Guanghua
Published: June 2021

e14021

Background: The updated 2016 edition of the WHO Classification of CNS tumors indicates that IDH1 R132H, H3 K27M mutations, and co-deletion of 1p19q are strong stratification and prognostic markers glioma. FISH/IHC, as the commonly detected methods, present specific false-negative rates in the actual condition. Methods: In our study, IDH1 R132H status of 158 cases was assessed by IHC and NGS, and H3 K27M statuses of 83 patients were evaluated by IHC and NGS. 22 positive cases of 1p/19q co-deletion, all confirmed by FISH, were assessed by NGS. Results: For IDH1 R132H, 2 cases were IHC negative and were positive as confirmed by NGS. Another 10 patients with weakly IHC positive results were negative in NGS. Combined with histologic hallmarks, 6 cases of these samples could be diagnosed as glioblastoma, IDH wildtype; 1 case with POLE could be diagnosed as giant cell glioblastoma; 3 cases with BRAF V600E mutation, BRAF fusion, and ATRX mutation, respectively, could be diagnosed as pilocytic astrocytoma. Towards H3 K27M, 3 cases with IHC weakly positive were negative in NGS. Among these samples, 2 cases were diagnosed as glioblastoma, IDH wildtype by molecular and histologic hallmarks, and 1 case was medulloblastoma, SHH. Using NGS, IDH1 R132H /H3 K27M statues can be distinctly distinguished in which that is unknown by IHC. The results of NGS and FISH showed a 90.9%(20/22) consistent rate for the 1p19q co-deletion. 1 case was 1p deletion and intact 19q by FISH while 1p19q co-deletion by NGS because of the 19p deletion. 1 case was 1p19q co-deletion by FISH but 1p19q wildtype by NGS, diagnosed as glioblastoma, IDH wildtype with chr7+/10-. Conclusions: In our study, the agreement between NGS results and clinical pathology diagnosis was approximately 100%. NGS may act as the primary technology of molecular classification in glioma in the future.
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.
INTASYL self-delivering RNAi therapy specifically dual-targeting BRD4 and PD-1 to elicit complete tumor responses and evidence of synergy in a subcutaneous Hepa1-6 model of murine hepatoma in C57BL/6N mice.
Authors: Cuiffo Benjamin , Maxwell Melissa , Yan Dingxue , Rivest Brianna , Cardia James , Fricker Simon P
Published: June 2021

e14537

Background: The therapeutic potential of targeting bromodomain and extra-terminal motif (BET) protein BRD4 is supported by promising preclinical data in multiple cancer contexts. However, clinical trials of BET inhibitors (BETi) have overall shown limited clinical efficacy and only passable safety profiles as monotherapy. BET proteins are now also understood to control a wide range of immunoregulatory pathways. Preclinical data suggests a synergistic impact with immune checkpoint blockade (ICB) targeting PD-1. Trials assessing the potential of BETi in rational combination with ICB are underway. The INTASYL platform is a self-delivering RNAi technology enabling efficient delivery into target cells without need for specialized drug delivery systems. INTASYL provides specific, robust, and durable on-target gene silencing. Furthermore, INTASYL moieties targeting multiple specific targets can be formulated together, providing a highly versatile platform for providing multi-targeted combination therapy in a single drug substance. Methods: Here we assessed the efficacy of mouse/human BRD4-targeting INTASYL PH-894 as monotherapy or co-formulated with mouse PD-1 targeting INTASYL mPH-762, in treating a subcutaneous Hepa1-6 model of murine hepatoma in C57BL/6N mice. On-target, specific silencing of BRD4 and PD-1 was demonstrated in vitro. In vivo, mice bearing tumors of ̃150 mm3 (N = 12/arm) were treated intratumorally on Days 1, 3, 7, 10 & 14 with of PH-894 or mPH-762 as single-agent, or PH-894/mPH-762 in combination at sub-optimal doses (0.1 mg/INTASYL/dose), or with PH-894/mPH-762 at half-suboptimal dose (0.05mg/INTASYL/dose) to assess potential synergistic impacts. Positive control groups received systemic (IP) (+)-JQ-1 (13.33mg/dose; qdx5), anti-mouse PD-1 antibody (75 µg; Days 1, 3, 7, 10, 14), or both treatments. Negative control animals received vehicle only. Tumor volumes and body weights were recorded longitudinally. Results: Compared to vehicle, suboptimal (+)-JQ-1, anti-PD-1, PH-894 or mPH-762 each elicited tumor control but not resolution as expected; while treatment with suboptimal PH-894/mPH-762 combo produced complete response (CR) in 75% (9/12) mice, outperforming combination treatment with (+)-JQ-1 and anti-PD-1 mAb [CR 58% (7/12) of mice]. Treatment with half-suboptimal doses of PH-894/mPH-762 resulted in CR for 42% (5/12) mice and tumor control exceeding that elicited by each monotherapy, suggesting potential synergy of the dual-targeting formulation. Tumors were isolated for mechanistic analyses of immune and tumor cells. Conclusions: These data demonstrate proof-of-concept of the therapeutic potential of rational dual-targeted INTASYL provided as single formulation in vivo, suggesting potential for meaningful clinical therapeutic impact.
Nephrotoxicity in patients with cancer treated with immune checkpoint inhibitors.
Authors: Eleutherakis Papaiakovou Evangelos , Kastritis Efstathios , Zagouri Flora , Grapsa Dimitra , Ntanassis-Stathopoulos Ioannis , Stefanou Dimitra , Evangelou Georgios , Kotteas Ilias , Dimopoulos Meletios A. , Syrigos Konstantinos
Published: June 2021

e14558

Background: Despite their efficacy in various cancer types, administration of immune checkpoint inhibitors (ICIs) may be complicated by the development of immune-related adverse events (IrAE). Nephrotoxicity represents a relatively rare but clinically significant adverse event, associated with the administration of ICI. Methods: The medical records of 294 patients with solid tumors or hematological malignancies that received ICIs for at least 4 weeks were retrospectively reviewed. Demographic, clinicopathological, treatment, toxicity and outcome data were recorded. ICI-associated nephrotoxicity was correlated with the remaining clinicopathological parameters as well as with progression-free survival (PFS) and overall survival (OS). Results: The majority of patients were male (70.4%), with bladder cancer or renal cell carcinoma (38.1% and 27.6%, respectively), and stage IV disease (56.8%), and had received ICI as monotherapy (68,7%). Nephrotoxicity during immunotherapy administration was observed in 20/ 294 (6.8%) patients, was mild (grade 1) and reversible in 55% and 75% of these cases, respectively, and was significantly associated with nephrotoxicity from previously administered therapy (p = 0,008), impaired renal function at initiation of ICIs (p = 0.044). and advanced disease stage at diagnosis (p = 0,028). No correlation between ICI-nephrotoxicity and PFS or OS was observed. Conclusions: ICI-associated nephrotoxicity was a relatively infrequent IrAE in our patient population, most commonly mild and reversible, and with no significant effect on survival. Preexisting treatment-related nephrotoxicity, impaired renal function and/or advanced disease stage at diagnosis may represent significant predisposing factors for its development.
Repurposing infectious diseases vaccines against cancer.
Authors: Bouche Gauthier , Vandeborne Liese , Pantziarka Pan , Van Nuffel An M. T.
Published: June 2021

e14564

Background: Vaccines used to prevent infection have long been known to stimulate immune responses to cancer as illustrated by the approval of the Bacillus Calmette–Guérin (BCG) vaccine to treat bladder cancer since the 1970s. The recent approval of immunotherapies has rejuvenated this research area with reports of anti-tumour responses with existing infectious diseases vaccines used as such, either alone or in combination with approved immune checkpoint inhibitors. This prompted us to review recent research activities using approved vaccines to treat cancer. Methods: PubMed and clinicaltrials.gov databases were queried for all 31 infectious diseases having at least one preventive vaccine approved, and all hits were reviewed for relevance. Only articles and trial registrations about infectious diseases vaccines used in an unmodified form as treatment for cancer, were included. The search was restricted in time, capturing articles from the year 2000 until November 2020. Results: A total of 1,754 PubMed abstracts and 549 trial registrations were screened. Data supporting a cancer therapeutic use was found for 16 of the 31 infectious diseases vaccines. For 10 (BCG, diphtheria, tetanus, human papillomavirus (HPV), influenza, measles, pneumococcus, smallpox, typhoid and varicella-zoster), clinical trials have been conducted or are ongoing. Within the remaining 6, preclinical evidence supports further evaluation of the rotavirus, yellow fever and pertussis vaccine in carefully designed clinical trials. The mechanistic evidence for the cholera vaccine, combined with the observational data in colorectal cancer, is also supportive of clinical translation. There is limited data for the hepatitis B vaccine and for the mumps vaccine alone. Four findings are worth highlighting: first, the superiority of intra-vesical typhoid vaccine instillations over BCG instillations in a preclinical bladder cancer model, which is now the subject of a phase I trial; second, the perioperative use of the influenza vaccine to limit and prevent the NK cell dysfunction induced by cancer surgery; third, objective responses following intra-tumoral injections of the measles vaccine in cutaneous T-cell lymphoma; fourth, objective responses induced by the HPV vaccine in skin squamous cell carcinoma. Conclusions: Sixteen infectious diseases vaccines have been or are currently being explored for repurposing in oncology. All vaccines are intended to induce or improve an anti-tumour (immune) response. Next to biological and immunological mechanisms, that are very different from one vaccine to another, also the mode of administration and combinations and sequence with other (immuno-)therapies deserve more attention in future research.
Association of immune checkpoint inhibitors (ICI) and venous thromboembolism (VTE): A single healthcare system experience.
Authors: Ananthula Aneesha , Chapple Andrew G , Mani Shyam , Nguyen Anh , Del Toro Alejandra Matilde , Boulmay Brian C.
Published: June 2021

e14567

Background: Cancer and some anti-cancer therapies increase risk of VTE. ICIs cause immune activation and inflammation which are known to potentiate VTE formation. The risk of VTE associated with ICI is unclear, with conflicting evidence of association. Our study assessed the risk of VTE in patients treated with ICI compared to those receiving other cancer treatments. Methods: EPIC Slicer Dicer was used to collect data from three university-affiliated hospitals in New Orleans, LA. All patients >17 years of age, with an oncology treatment plan (TP) between January 1 2012 – January 31 2021 were considered the base population (n=6,894) for the study. The base population was divided into two cohorts - those that received ICI (n=794) versus those who did not (n=6,100). ICIs included pembrolizumab, nivolumab, avelumab, atezolizumab, durvalumab, cemiplimab, ipilimumab. An encounter diagnosis of deep venous thrombosis (DVT) or pulmonary embolism (PE) which was preceded by a TP in the last 1 year was used to identify patients with VTE after initiation of a TP. Categorical covariates compared across VTE groups using Chi-squared tests of independence. Two sample t-tests were used to compare continuous covariates based on VTE status. Logistic regression used to predict VTE as a function of ICIs and other potential confounders including age, BMI, gender, race and cancer subtypes. Results: Patients who experienced VTE were significantly more likely to have received ICI (1.43 times odds of VTE (95% CI = 1.12-1.83; p <0.005), be black or African American (p < 0.05 compared with White and Asian races), and have GI cancer (1.82 times odds, 95% CI= 1.41 – 2.36, p< 0.001), GU cancer (1.7 times odds, 95% CI = 1.3 – 2.23; p<0.001). Patients with VTE were significantly less likely to have breast cancer or other cancer types. Males had a decreased odd of VTE (0.73 odds, 95% CI= 0.62 – 0.85, p< 0.001). Conclusions: Based on the above results, there appears to be an increased likelihood of VTE in patients that receive ICI when compared to other types of cancer treatment. This study is a preliminary analysis and has limitations. At this time, it is unclear if prophylactic anticoagulation can reduce risk of VTE after ICI initiation. Further investigation is warranted in the form of individual chart review, or a prospective study.
Six-gene model based on expression predicts distinct disease-free survival in early-stage microsatellite stability (MSS) endometrial, colorectal and stomach tumors.
Authors: Chen Tufeng , Liu Jianpei , Liu Xinyi , Huang Mengli
Published: June 2021

e14578

Background: Microsatellite stability (MSS) tumors hardly benefit from immunotherapies and are more probable to occur postoperative recurrence. However, some studies have revealed that a subset of MSS patients harbor “hot immune microenvironment” tumors, indicating high heterogeneity in such wide range of patient population. On the other hand, researches of mechanism of MSI formation found potential similarities in endometrial and gastrointestinal tumors. We hypothesized the transcriptomic features in these cancers correlated with immune-related signatures and patients’ prognosis. Methods: Early stage (I-III stage) MSS tumors, including endometrial, colorectal, and gastric from TCGA project were analyzed as training cohort (n=170). A combined cohort consisting of 604 colorectal and stomach cancers from GEO datasets (GSE39582，GSE62254) was validation cohort. The RNA-Seq profiling data and disease-free survival (DFS) data of patients were collected. Cibersort tool was used to evaluate twenty-two immune cells’ enrichment. The prediction model was developed by three steps: Univariate cox regression of DFS was conducted to select 9 immune cells. Then the train cohort was divided into two groups based on non-negative matrix factorization (NMF) method using this 9 immune cell features. Differentially expressed genes of these two groups were identified and screened further by lasso regression. Log-rank test was used to evaluate the difference of DFS. Results: A six-gene lasso-cox model was developed. The genes were LYZ, WFDC2, CAPS, RHPN1, TFF2 and TGFBR2. Based on the score evaluated by this model, patients in training cohort were divided into high-risk and low-risk groups. Low-risk population had much longer DFS (HR 0.07, 95%CI 0.03-0.18, p<0.001). In validation cohort, lower risk score was also verified to be associated with a lower likelihood of recurrence (HR 0.66, 95%CI 0.5-0.88, p=0.0047). Conclusions: We developed a model of six-genes predicting disease-free survival based on RNA-Seq data in early stage MSS patients. Further validation was needed to implement in larger clinical cohorts.
The landscape of intergenic-breakpoint fusion in lung cancer.
Authors: Liang Hua , Wu Yaqing , Luo Ningning , Zhang Qin , Qi Yingxue , Duan Qianqian , Qi Chuang
Published: June 2021

e15002

Background: Kinase fusions have been proved as driver factors and drug targets for lung cancer. DNA-based next-generation sequencing (NGS) has been widely used for gene fusion detection in patients with lung cancer and intergenic-breakpoint fusions can be detected, in which breakpoints localize to intergenic regions. Theoretically, intergenic-breakpoint fusions are unlikely to be functional, but recent studies have found that intergenic-breakpoint fusions are potentially activated. However, the proportion of intergene-breakpoint fusion occurrence in lung cancer is still unclear. Hereby, next generation sequencing was performed to describe lung intergenic-breakpoint fusions for further research. Methods: Tumor tissue, pleural effusion, or blood samples of 6798 patients with lung cancer from Simceredx were collected for targeted NGS panel sequencing from June 2020 to January 2021. Gene rearrangements of ALK, ROS1, RET, NTRK(1-3) were assessed. Results: We found gene fusions in 4.43% (301/6798) patients with lung cancer totally, of which 59.1% (178/301) ALK fusions, 18.9% (57/301) RET fusions, 15.9% (48/301) ROS1 fusions and 6.64% (20/301) NTRK(1-3) fusions. The intergenic-breakpoint fusions accounted for all fusions of ALK, RET and ROS1 genes were 24.2% (43/178), 22.8% (13 /57) and 18.8% (9/48), respectively. In patients with NTRK intergenic fusions (50.0%, 10/20), NTRK1, NTRK2 and NTRK3 was involved in 2, 1, 7, respectively. Co-occurrence of different fusions was rare. Conclusions: Our data firstly revealed the specific intergenic-breakpoint fusions ( ALK, RET, ROS1, NTRK1-3) in lung cancer. The functional consequences identifying by RNA or protein assay and clinical validation need to be carried out in the future.
Deep learning to identify and predict cardiotoxicities of anticancer drugs.
Authors: Sarangdhar Mayur , Kolli Venkatesh , Seibel William , Perentesis John Peter
Published: June 2021

e15012

Background: Recent advances in cancer treatment have revolutionized patient outcomes. However, toxicities associated with anti-cancer drugs remain a concern with many anti-cancer drugs now implicated in cardiotoxicity. The complete spectrum of cardiotoxicity associated with anti-cancer drugs is only evident post-approval of drugs. Deep Learning methods can identify novel and emerging safety signals in “real-world” clinical settings. Methods: We used AERS Mine, an open-source data mining platform to identify drug toxicity signatures in the FDA’s Adverse Event Reporting System of 16 million patients. We identified 1.3 million patients on traditional and targeted anti-cancer therapy to analyze therapy-specific cardiotoxicity patterns. Cardiotoxicity training dataset contained 1571 molecules characterized with bioassay against hERG potassium channel and included 350 toxic compounds with an IC50 of < 1μM. We implemented a Deep Belief Network to extract a deep hierarchical representation of the training data, and the Extra Tree Classifier to predict the toxicity of drug candidates. Drugs were encoded using 1024-bit Morgan fingerprint representation using SMILES with search radius of 7 atoms. Pharmacovigilance metrics (Relative Risks and safety signals) were used to establish statistical correlation. Results: This analysis identified signatures of arrhythmias and conduction abnormalities associated with common anti-cancer drugs (e.g. atrial fibrillation with ibrutinib, alkylating agents, immunomodulatory drugs; sinus bradycardia with 5FU, paclitaxel, thalidomide; sinus tachycardia with anthracyclines). Our analysis also identified myositis/myocarditis association with newer immune checkpoint inhibitors (e.g., atezolizumab, durvalumab, cemiplimab, avelumab) paralleling earlier signals for pembrolizumab, nivolumab, and ipilimumab. Deep Learning identified signatures of chemical moieties linked to cardiotoxicity, including common motifs in drugs associated with arrhythmias and conduction abnormalities with an accuracy of 89%. Conclusions: Deep Learning provides a comprehensive insight into emerging cardiotoxicity patterns of approved and investigational drugs, allows detection of ‘rogue’ chemical moieties, and shows promise for novel drug discovery and development.