Galectin-9/TIM-3 as a Key Regulator of Immune Response in Gliomas With Chromosome 1p/19q Codeletion

Gliomas with chromosome 1p/19q codeletion were considered a specific tumor entity. This study was designed to reveal the biological function alterations tightly associated with 1p/19q codeletion in gliomas. Clinicopathological and RNA sequencing data from glioma patients were obtained from The Cancer Genome Atlas and Chinese Glioma Genome Atlas databases. Gene set variation analysis was performed to explore the differences in biological functions between glioma subgroups stratified by 1p/19q codeletion status. The abundance of immune cells in each sample was detected using the CIBERSORT analytical tool. Single-cell sequencing data from public databases were analyzed using the t-distributed stochastic neighbor embedding (t-SNE) algorithm, and the findings were verified by in vitro and in vivo experiments and patient samples.We found that the activation of immune and inflammatory responses was tightly associated with 1p/19q codeletion in gliomas. As the most important transcriptional regulator of Galectin-9 in gliomas, the expression level of CCAAT enhancer-binding protein alpha in samples with 1p/19q codeletion was significantly decreased, which led to the downregulation of the immune checkpoints Galectin-9 and TIM-3. These results were validated in three independent datasets. The t-SNE analysis showed that the loss of chromosome 19q was the main reason for the promotion of the antitumor immune response. IHC protein staining, in vitro and in vivo experiments verified the results of bioinformatics analysis. In gliomas, 1p/19q codeletion can promote the antitumor immune response by downregulating the expression levels of the immune checkpoint TIM-3 and its ligand Galectin-9.

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RBM8A Promotes Glioblastoma Growth and Invasion Through the Notch/STAT3 Pathway

Frontiers in Oncology ◽

10.3389/fonc.2021.736941 ◽

2021 ◽

Vol 11 ◽

Author(s):

Yan Lin ◽

Lei Wei ◽

Beiquan Hu ◽

Jinyan Zhang ◽

Jiazhang Wei ◽

...

Keyword(s):

Cell Proliferation ◽

Rna Binding ◽

The Cancer Genome Atlas ◽

Biological Behavior ◽

Binding Motif ◽

Sequencing Data ◽

Stat3 Pathway ◽

Genome Atlas

BackgroundGlioblastoma (GBM) is a prevalent brain malignancy with an extremely poor prognosis, which is attributable to its invasive biological behavior. The RNA-binding motif protein 8A (RBM8A) has different effects on various human cancers. However, the role of RBM8A in GBM progression remains unclear.MethodsWe investigated the expression levels of RBM8A in 94 GBM patients and explored the correlation between RBM8A expression and patient prognosis. Using in vitro and in vivo assays, combined with GBM sequencing data from the Cancer Genome Atlas (TCGA) and the Chinese Glioma Genome Atlas (CGGA), we examined whether and how RBM8A contributes to GBM progression.ResultsRBM8A was up-regulated in GBM tissues, and its higher expression correlated with worse prognosis. Knockdown of RBM8A inhibited GBM progression and invasion ability both in vitro and in vivo. On the contrary, overexpression of RBM8A promoted GBM progression and invasion ability. Enrichment analysis of differentially expressed genes in GBM data identified the Notch1/STAT3 network as a potential downstream target of RBM8A, and this was supported by molecular docking studies. Furthermore, we demonstrated that RBM8A regulates the transcriptional activity of CBF1. The γ-secretase inhibitor DAPT significantly reversed RBM8A-enhanced GBM cell proliferation and invasion, and was associated with down-regulation of p-STAT3 and Notch1 protein. Finally, the gene set variance analysis score of genes involved in regulation of the Notch1/STAT3 network by RBM8A showed good diagnostic and prognostic value for GBM.ConclusionsRBM8A may promote GBM cell proliferation and migration by activating the Notch/STAT3 pathway in GBM cells, suggesting that RBM8A may serve as a potential therapeutic target for the treatment of GBM.

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EXTH-45. MICROGLIA-SPECIFIC DISRUPTION OF SIALIC ACID-SIGLEC-9/E INTERACTIONS: A NOVEL IMMUNOTHERAPY AGAINST GLIOBLASTOMA?

Neuro-Oncology ◽

10.1093/neuonc/noab196.684 ◽

2021 ◽

Vol 23 (Supplement_6) ◽

pp. vi173-vi173

Author(s):

Philip Schmassmann ◽

Tomás A Martins ◽

Michal Stanczak ◽

Marie-Françoise Ritz ◽

Tala Shekarian ◽

...

Keyword(s):

Immune Response ◽

Sialic Acid ◽

Therapeutic Potential ◽

Adaptive Immune Response ◽

Cell Uptake ◽

Immune Checkpoints ◽

Sequencing Data ◽

Ki 67 ◽

Spatio Temporal

Abstract Recently, ‘don’t eat me’-signals like CD47 have emerged as novel innate immune checkpoints, enabling cancer cells to evade clearance by phagocytes such as monocyte-derived macrophages (MDM) or microglia (MG). Here, we aim at defining the role of inhibitory Siglec-9 in human and its mouse homologue Siglec-E in MG-centered immunotherapy against GBM. TCGA RNA-sequencing data revealed a significant correlation between high expression of immunoinhibitory SIGLEC9 and poor survival in GBM patients (log-rank p = 0.02). Siglec-E blockade increased murine MG mediated GBM cell in vitro phagocytosis (normalized phagocytosis of 1.00 in isotype vs. 1.76 in anti-Siglec-E antibody, p < 0.001). By employing a CT-2A orthotopic GBM mouse model with MG-specific spatio-temporal deletion of Siglece (Sall1 CreERT2 x Siglece flox ), we observed high MG-proliferation upon Siglec-E knockdown (Ki-67+ MG 14.8% in Cre- vs. 34.9% in Cre+, p < 0.0001) accompanied by an enhanced microglial GBM-cell uptake (5.6% in Cre- vs. 12.3% in Cre+, p < 0.001). This beneficial response was counteracted by an accentuated influx of pro-tumorigenic MDM in the Cre+ group (CD163high CD86low MDM of total MDM 47.1% in Cre- vs. 65.3% in Cre+, p = 0.002), which prevented an efficient adaptive anti-tumor immune response and survival benefit. Currently, we are investigating the cross-talk between GBM-associated MG and MDM upon Siglec-E knockdown by scRNAseq of the tumor-infiltrating immune compartment, including TCR-clonotype tracking. By genetically targeting sialic acids, the ligand for Siglec receptors, on CT-2A cells (GNE-KO), we observed a strong innate and adaptive immune response with less exhausted tumor-infiltrating CD8+ T-cells (14.8% in WT vs. 5.9% in GNE-KO, p = 0.003), which resulted in a prolonged survival (30d in WT vs. 41d post-tumor-injection in GNE-KO, p = 0.03). These data identify the sialic-acid-Siglec-E pathway as an anti-phagocytic signal in a pre-clinical GBM model, and demonstrate its therapeutic potential in GBM immunotherapy.

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Immune and clinical features of CD96 expression in glioma by large-scale analysis

10.21203/rs.2.23546/v1 ◽

2020 ◽

Author(s):

Qiang Zhang ◽

Hua Zhong ◽

Yinchun Fan ◽

Qian Liu ◽

Jiancheng Song ◽

...

Keyword(s):

Immune Response ◽

T Cells ◽

Large Scale ◽

The Cancer Genome Atlas ◽

Immune Checkpoints ◽

Immune Functions ◽

Clinical Prognosis ◽

Regulatory Pathways ◽

Cox Analysis ◽

Genome Atlas

Abstract Background: Immune checkpoints target regulatory pathways in T cells which enhance antitumor immune responses and elicit durable clinical responses . As a novel immune checkpoint, CD96 is an attractive key target for cancer immunotherapy. However, there is no integrative investigation of CD96 in glioma. Our study explored the relationship between CD96 expression and clinical prognosis in glioma. Methods: A total of 1,024 RNA and clinical data were enrolled in this study, including 325 samples from the Chinese Glioma Genome Atlas (CGGA) database and 699 samples from The Cancer Genome Atlas (TCGA) dataset. R language was used to perform statistical analysis and draw figures. Results: CD96 had a consistently positive relationship with glioblastoma and highly enriched in IDH-wildtype and mesenchymal subtype glioma. GO enrichment and GSVA analyses suggested that CD96 was more involved in immune functions, especially related to T cell-mediated immune response in glioma. Subsequent immune infiltration analysis manifes ted that CD96 was positively correlated with infiltrating levels of CD4+ T and CD8+ T cells, macrophages , neutrophils, and DCs in GBM and LGG. Additionally, CD96 was tightly associated with other immune checkpoints including PD-1 , CTLA-4 , TIGIT , and TIM-3 . Univariate and multivariate Cox analysis demonstrated that CD96 acts as an independent indicator of poor prognosis in glioma. Conclusion: CD96 expression was increased in malignant phenotype and negatively associated with overall survival (OS) in glioma. CD96 also showed a positive correlation with other immune checkpoints, immune response, and inflammatory activity. Our findings indicate that CD96 is a promising clinical target for further immunotherapeutic in glioma patients.

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Angiostatic activity of the antitumor cytokine interleukin-21

Blood ◽

10.1182/blood-2007-09-113878 ◽

2008 ◽

Vol 112 (13) ◽

pp. 4940-4947 ◽

Cited By ~ 36

Author(s):

Karolien Castermans ◽

Sebastien P. Tabruyn ◽

Rong Zeng ◽

Judy R. van Beijnum ◽

Cheryl Eppolito ◽

...

Keyword(s):

Immune Response ◽

Tumor Angiogenesis ◽

Chorioallantoic Membrane ◽

Antitumor Immune Response ◽

In Vivo Studies ◽

Type I ◽

Stat3 Phosphorylation ◽

Interleukin 21

Abstract Interleukin-21 (IL-21) is a recently described immunoregulatory cytokine. It has been identified as a very potent immunotherapeutic agent in several cancer types in animal models, and clinical studies are ongoing. IL-21 belongs to the type I cytokine family of which other members, ie, IL-2, IL-15, and IL-4, have been shown to exert activities on vascular endothelial cells (ECs). We hypothesized that IL-21, in addition to inducing the antitumor immune response, also inhibits tumor angiogenesis. In vitro experiments showed a decrease of proliferation and sprouting of activated ECs after IL-21 treatment. We found that the IL-21 receptor is expressed on vascular ECs. Furthermore, in vivo studies in the chorioallantoic membrane of the chick embryo and in mouse tumors demonstrated that IL-21 treatment disturbs vessel architecture and negatively affects vessel outgrowth. Our results also confirm the earlier suggested angiostatic potential of IL-2 in vitro and in vivo. The angiostatic effect of IL-21 is confirmed by the decrease in expression of angiogenesis-related genes. Interestingly, IL-21 treatment of ECs leads to a decrease of Stat3 phosphorylation. Our research shows that IL-21 is a very powerful antitumor compound that combines the induction of an effective antitumor immune response with inhibition of tumor angiogenesis.

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Local Delivery of Minocycline and Vorinostat Act Synergistically to Inhibit Recurrence of Gliomas

10.21203/rs.3.rs-33885/v1 ◽

2020 ◽

Author(s):

Gang Zhao ◽

Jun Jia ◽

Lansheng Wang ◽

Yongkang Zhang ◽

Han Yang ◽

...

Keyword(s):

High Performance ◽

Postoperative Recurrence ◽

The Cancer Genome Atlas ◽

The Public ◽

Continuous Release ◽

Clinical Consequences ◽

Cancer Genome Atlas ◽

Genome Atlas

Abstract Background:Postoperative recurrence is the main reason of poor clinical consequences in glioma patients, so preventing recurrence of tumors is crucial in management of gliomas. Methods:In this study, the expression of matrix metalloproteinases (MMPs)in tissues from normal were detected by using RNA-seq analysis.Glioma cases from the public databases (The Cancer Genome Atlas (TCGA), The Chinese Glioma Genome Atlas(CGGA), Betastasis) were included in this study.The hydrogelcontains minocycline (Mino) and vorinostat (Vor)(G/Mino + Vor) was formed under 365 nm when photoinitiator was added in. High Performance Liquid Chromatography (HPLC) assay was used to assessed the release of drugs in G/Mino + Vor hydrogel. MTT assay was used to explore the biosecurity of GelMA. Immunohistochemistry assay, ELISA assay, Tunel assay were used to demonstrate the antitumor effect of G/Mino + Vor hydrogel.Results:We developed G/Mino + Vor hydrogel successfully. Thenthe experiment in vitro and in vivo confirmed MMPs-responsive delivery of minocycline and vorinostat in hydrogel and the anti-glioma effect on incomplete tumor operation model, which indicated that G/Mino + Vor hydrogel effectively inhibited the recurrence of glioma after surgery.Conclusions: In summary, G/Mino + Vor hydrogel could continuous release minocycline and vorinostat in surgical cavity for inhibiting local recurrence of glioma after operation.

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Capsular polysaccharide correlates with immune response to the human gut microbe Ruminococcus gnavus

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.2007595118 ◽

2021 ◽

Vol 118 (20) ◽

pp. e2007595118

Author(s):

Matthew T. Henke ◽

Eric M. Brown ◽

Chelsi D. Cassilly ◽

Hera Vlamakis ◽

Ramnik J. Xavier ◽

...

Keyword(s):

Immune Response ◽

Molecular Mechanisms ◽

Capsular Polysaccharide ◽

Inflammatory Responses ◽

Cell Wall Polysaccharide ◽

Proinflammatory Responses ◽

Gut Microbe ◽

Inflammatory Bowel

Active inflammatory bowel disease (IBD) often coincides with increases of Ruminococcus gnavus, a gut microbe found in nearly everyone. It was not known how, or if, this correlation contributed to disease. We investigated clinical isolates of R. gnavus to identify molecular mechanisms that would link R. gnavus to inflammation. Here, we show that only some isolates of R. gnavus produce a capsular polysaccharide that promotes a tolerogenic immune response, whereas isolates lacking functional capsule biosynthetic genes elicit robust proinflammatory responses in vitro. Germ-free mice colonized with an isolate of R. gnavus lacking a capsule show increased measures of gut inflammation compared to those colonized with an encapsulated isolate in vivo. These observations in the context of our earlier identification of an inflammatory cell-wall polysaccharide reveal how some strains of R. gnavus could drive the inflammatory responses that characterize IBD.

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ANXA2 is Correlated With the Molecular Features and Clinical Prognosis of Glioma, and Acts as a Potential Marker of Immunosuppression

10.21203/rs.3.rs-665909/v1 ◽

2021 ◽

Author(s):

Kaiming Ma ◽

Xin Chen ◽

Weihai Liu ◽

Yang Yang ◽

Suhua Chen ◽

...

Keyword(s):

Immune Response ◽

Suppressor Cells ◽

High Grade Glioma ◽

The Cancer Genome Atlas ◽

Sequencing Data ◽

Clinical Value ◽

Clinical Prognosis ◽

Molecular Features ◽

Potential Marker ◽

Genome Atlas

Abstract Background: Recent studies have shown that ANXA2 plays a crucial role in the development of many cancers, while its role in glioma remains unclear. Thus, our study aimed to investigate the molecular biological characteristics and clinical value of ANXA2 in glioma. Methods: We analyzed the RNA sequencing data of 325 glioma samples from the Chinese Glioma Genome Atlas (CGGA) as the training cohort and the RNA expression data of 699 samples from The Cancer Genome Atlas (TCGA) database as the validation cohort, totaling approximately 1024 samples. Results: We found that ANXA2 was overexpressed significantly in high-grade glioma (HGG), isocitrate dehydrogenase (IDH) wild-type glioma and mesenchymal-subtype glioma. Receiver operating characteristic (ROC) analysis revealed that ANXA2 was a potential indicator of the mesenchymal subtype. Kaplan–Meier curve analysis revealed that high expression of ANXA2 indicated significantly poorer survival in glioma and HGG patients. Multivariate Cox analysis also demonstrated that ANXA2 acted as an independent indicator of poor prognosis in glioma patients. Further gene ontology (GO) analysis showed that ANXA2-related genes were mainly involved in the innate immune response and inflammatory response of glioma, and these genes were positively correlated with the expression of ANXA2. Then, we selected seven immune-related meta-genes and found that ANXA2 was positively correlated with HCK, LCK, MHC II, STAT1 and interferon but negatively correlated with IgG. Subsequent correlation analysis of infiltrating immune cells showed that ANXA2 was positively correlated with the infiltration of tumor-related macrophages (TAMs), regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs), suggesting that ANXA2 may be involved in the immunosuppression of glioma.Conclusion: Our study revealed that ANXA2 is closely related to the malignant glioma phenotype and poor patient prognosis. ANXA2 also plays an important role in the immune response and inflammatory activity and can be considered a biomarker of immunosuppression in glioma. These findings suggest that ANXA2 may be a promising target for the further development of immunotherapeutic for glioma.

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RUNX1 contributes to the mesenchymal subtype of glioblastoma in a TGFβ pathway-dependent manner

Cell Death and Disease ◽

10.1038/s41419-019-2108-x ◽

2019 ◽

Vol 10 (12) ◽

Cited By ~ 6

Author(s):

Kai Zhao ◽

Xiaoteng Cui ◽

Qixue Wang ◽

Chuan Fang ◽

Yanli Tan ◽

...

Keyword(s):

Target Genes ◽

The Cancer Genome Atlas ◽

Luciferase Reporter ◽

Dependent Manner ◽

Functional Studies ◽

The Impact ◽

Tgfβ Pathway ◽

Genome Atlas

AbstractRunt-Related Transcription Factor 1 (RUNX1) is highly expressed in the Mesenchymal (Mes) subtype of glioblastoma (GBM). However, the specific molecular mechanism of RUNX1 in Mes GBM remains largely elusive. In this study, cell and tumor tissue typing were performed by RNA-sequencing. Co-immunoprecipitation (co-IP) and immunofluorescence (IF) were employed to identify members of the RUNX1 transcriptional protein complex. Bioinformatics analysis, chromatin immunoprecipitation (ChIP), and luciferase reporter experiments were utilized to verify target genes. Analyses of The Cancer Genome Atlas (TCGA) and Chinese Glioma Genome Atlas (CGGA) verified the expression levels and prognoses associated with RUNX1/p-SMAD3/SUV39H1 target genes. In vivo patient-derived xenograft (PDX) studies and in vitro functional studies verified the impact of RUNX1 on the occurrence and development of GBM. The results showed that RUNX1 was upregulated in Mes GBM cell lines, tissues and patients and promoted proliferation and invasion in GBM in a TGFβ pathway-dependent manner in vivo and in vitro. We found and verified that BCL3 and MGP are transcriptionally activated by p-SMAD3 /RUNX1, while MXI1 is transcriptionally suppressed by the RUNX1/SUV39H1-H3K9me3 axis. This finding offers a theoretical rationale for using molecular markers and choosing therapeutic targets for the Mes type of GBM.

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SRSF6-regulated alternative splicing that promotes tumour progression offers a therapy target for colorectal cancer

Gut ◽

10.1136/gutjnl-2017-314983 ◽

2017 ◽

Vol 68 (1) ◽

pp. 118-129 ◽

Cited By ~ 30

Author(s):

Ledong Wan ◽

Wenying Yu ◽

Enhui Shen ◽

Wenjie Sun ◽

Yuan Liu ◽

...

Keyword(s):

Colorectal Cancer ◽

Alternative Splicing ◽

De Novo ◽

Tumour Progression ◽

The Cancer Genome Atlas ◽

Chronic Obstructive ◽

Binding Motif ◽

Sequencing Data

ObjectiveTo investigate the molecular function of splicing factor SRSF6 in colorectal cancer (CRC) progression and discover candidate chemicals for cancer therapy through targeting SRSF6.DesignWe performed comprehensive analysis for the expression of SRSF6 in 311 CRC samples, The Cancer Genome Atlas and Gene Expression Omnibus (GEO) database. Functional analysis of SRSF6 in CRC was performed in vitro and in vivo. SRSF6-regulated alternative splicing (AS) and its binding motif were identified by next-generation RNA-sequencing and RNA immunoprecipitation sequencing (RIP-seq), which was validated by gel shift and minigene reporter assay. ZO-1 exon23 AS was investigated to mediate the function of SRSF6 in vitro and in vivo. Based on the analysis of domain-specific role, SRSF6-targeted inhibitor was discovered de novoby virtual screening in 4855 FDA-approved drugs and its antitumour effects were evaluated in vitroand in vivo.ResultsSRSF6 was frequently upregulated in CRC samples and associated with poor prognosis, which promoted proliferation and metastasis in vitro and in vivo. We identified SRSF6-regulated AS targets and discovered the SRSF6 binding motif. Particularly, SRSF6 regulates ZO-1 aberrant splicing to function as an oncogene by binding directly to its motif in the exon23. Based on the result that SRSF6 RRM2 domain plays key roles in regulating AS and biological function, indacaterol, a β2-adrenergic receptor agonist approved for chronic obstructive pulmonary disease treatment, is identified as the inhibitor of SRSF6 to suppress CRC tumourigenicity.ConclusionsSRSF6 functions the important roles in mediating CRC progression through regulating AS, and indacaterol is repositioned as an antitumour drug through targeting SRSF6.Accession numbersThe accession numbers for sequencing data are SRP111763 and SRP111797.

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Overexpression of RAB34 Associates With Tumor Aggressiveness and Immune Infiltration in Glioma

10.21203/rs.3.rs-779217/v1 ◽

2021 ◽

Author(s):

Peng Hou ◽

Quan Wan ◽

Qing Wang ◽

Xuechao Wu ◽

Xiaojie Lu

Keyword(s):

Epithelial To Mesenchymal Transition ◽

The Cancer Genome Atlas ◽

Immune Checkpoints ◽

Migration And Invasion ◽

Rna Seq ◽

Clinical Value ◽

Mesenchymal Transition ◽

Immune Infiltration ◽

Genome Atlas

Abstract Background: RAB34 is aberrantly expressed in various cancers and exhibits oncogenic properties. However, its function in glioma remains largely unclear. Herein, we investigated the clinical value and biological functions of RAB34 in glioma.Methods: In this study, we collected 697 RNA-seq data from The Cancer Genome Atlas (TCGA) dataset and 325 RNA-seq data from Chinese Glioma Genome Atlas (CGGA) dataset. CCK-8 and EdU assays were employed to assess cell proliferation ability. The transwell assay was utilized to explore cell migration and invasion capacities. Western blot coupled with qRT-PCR were employed to determine the protein, as well as RNA contents. The statistical analyses along with the graphical work were mainly implemented in the R software.Results: RAB34 expression was positively related to the glioma tumor grade, and predicted poor outcomes for glioma patients. RAB34 expression was significantly upregulated in classical and mesenchymal subtypes, and IDH wild-type gliomas. Additionally, RAB34 expression was regulated by promoter DNA methylation. Moreover, RAB34 expression was remarkably correlated with inflammatory activities, immune infiltration, and immune checkpoints in glioma. In vitro experiments demonstrated that inhibition of RAB34 restrained the growth, migration, as well as invasion of glioma cells, and reversed the epithelial-to-mesenchymal transition (EMT) process. Conclusion: Our findings established RAB34 as a novel progression-related biomarker and a possible immunotherapy target for glioma.

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