APOBEC3C is a novel target for the immune treatment of lower-grade gliomas

Apolipoprotein B mRNA editing catalytic polypeptide-like (APOBEC) type 3C (A3C) has been identified as a cancer molecular biomarker in the past decade. However, the practical role of A3C in lower-grade gliomas (LGGs) in improving the clinical outcome remains unclear. This study aims to discuss the function of A3C in immunotherapy in LGGs. The RNA-Sequencing (RNA-seq) and corresponding clinical data were extracted from UCSC Xena and the results were verified in the Chinese Glioma Genome Atlas (CGGA). Weighted gene co-expression network analysis (WGCNA) was used for screening A3C-related genes. Comprehensive bioinformation analyses were performed and multiple levels of expression, survival rate, and biological functions were assessed to explore the functions of A3C. A3C expression was significantly higher in LGGs than in normal tissues but lower than in glioblastoma (GBM), indicating its role as an independent prognosis predictor for LGGs. Twenty-eight A3C-related genes were found with WGCNA for unsupervised clustering analysis and three modification patterns with different outcomes and immune cell infiltration were identified. A3C and the A3C score were also correlated with immune cell infiltration and the expression of immune checkpoints. In addition, the A3C score was correlated with increased sensitivity to chemotherapy. Single-cell RNA (scRNA) analysis indicated that A3C most probably expresses on immune cells, such as T cells, B cells and macrophage. A3C is an immune-related prognostic biomarker in LGGs. Developing drugs to block A3C could enhance the efficiency of immunotherapy and improve disease survival. Abbreviation: A3C: Apolipoprotein B mRNA editing catalytic polypeptide-like (APOBEC) type 3C; LGGs: lower-grade gliomas; CGGA: Chinese Glioma Genome Atlas; WGCNA: Weighted gene co-expression network analysis; scRNA: Single-cell RNA; HGG: higher-grade glioma; OS: overall survival; TME: tumor microenvironment; KM: Kaplan-Meier; PFI: progression-free interval; IDH: isocitrate dehydrogenase; ROC: receiver operating characteristic; GS: gene significance; MM: module membership; TIMER: Tumor IMmune Estimation Resource; GSVA: gene set variation analysis; ssGSEA: single-sample gene-set enrichment analysis; PCA: principal component analysis; AUC: area under ROC curve; HAVCR2: hepatitis A virus cellular receptor 2; PDCD1: programmed cell death 1; PDCD1LG2: PDCD1 ligand 2; PTPRC: protein tyrosine phosphatase receptor type C; ACC: Adrenocortical carcinoma; BLCA: Bladder Urothelial Carcinoma;BRCA: Breast invasive carcinoma; CESC: Cervical squamous cell carcinoma and endocervical adenocarcinoma; CHOLCholangiocarcinoma; COADColon adenocarcinoma; DLBC: Lymphoid Neoplasm Diffuse Large B-cell Lymphoma; ESCA: Esophageal carcinoma; GBM: Glioblastoma multiforme; HNSC: Head and Neck squamous cell carcinoma; KICH: Kidney Chromophobe; KIRC: Kidney renal clear cell carcinoma; KIRP: Kidney renal papillary cell carcinoma; LAML: Acute Myeloid Leukemia; LGG: Brain Lower Grade Glioma; LIHC: Liver hepatocellular carcinoma; LUAD: Lung adenocarcinoma; LUSC: Lung squamous cell carcinoma; MESO: Mesothelioma; OV: Ovarian serous cystadenocarcinoma; PAAD: Pancreatic adenocarcinoma; PCPG: Pheochromocytoma and Paraganglioma; PRAD: Prostate adenocarcinoma; READ: Rectum adenocarcinoma; SARC: Sarcoma; SKCM: Skin Cutaneous Melanoma; STAD: Stomach adenocarcinoma; TGCT: Testicular Germ Cell Tumors; THCA: Thyroid carcinoma; THYM: Thymoma; UCEC: Uterine Corpus Endometrial Carcinoma; UCS: Uterine Carcinosarcoma; UVM: Uveal Melanoma</p

Stereological cell counts of 15G7 and TH immunohistochemistry.

<p>π… Data from the pilot study;</p>1<p>… Comparing TG vs. WT animals in TH IHC by Student's T Test;</p>2<p>… p<0.05 compared to 2 month old mice (Mann-Whitney U Test with Bonferroni correction for multiple comparisons);</p>3<p>… p<0.05 compared to 6 month old mice (Mann-Whitney U Test with Bonferroni correction for multiple comparisons). 15G7… human αSYN, TH… tyrosine hydroxylase, TG… transgenic, WT… wildtype, IHC… immunohistochemistry, SEM… standard error of mean.</p

Olfactory behavior.

<p>Mean investigation time in seconds in olfactory preference testing of 9 months old mice. Data are expressed as mean; error bars indicate the standard error of mean. Sample sizes are reported below the X-axis. * P<0.05.</p

Ferritin light chain expression in glioma and its correlation with survival in glioblastoma multiforme patients.

<p>(A) Ferritin light chain (FTL) expression levels were measured using real-time quantitative RT-PCR. Expression of FTL was significantly higher in the glioblastoma multiforme (GBM) group (grade IV) compared with the low-grade glioma group (grades I and II) (*P = 0.04). Data represent the mean ± standard deviation of triplicate experiments. (B and C) In silico analyses of overall survival and disease free survival of GBM patients according to their expression levels of FTL mRNA. Data were obtained from cBioPortal datasets [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0149361#pone.0149361.ref009" target="_blank">9</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0149361#pone.0149361.ref010" target="_blank">10</a>]. (D) In silico analysis showing overall survival of GBM patients according to their expression levels of FTH1 mRNA. Data were also obtained from the cBioPortal datasets [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0149361#pone.0149361.ref009" target="_blank">9</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0149361#pone.0149361.ref010" target="_blank">10</a>].</p

Ferritin light chain regulates the GADD45A/JNK pathway and interacts with GADD45A in glioblastoma multiforme cells.

<p>(A) Immunoblot analyses showing the effects of ferritin light chain (FTL) knockdown in U251 and A172 cells. Downregulation of FTL activated the GADD45A/JNK pathway in both cell lines, which was indicated by elevated levels of GADD45A, activation of JNKs (P < 0.01). Expression of Cyclin D1, c-myc, and PDGFR-β was also observed. The number at the top of each band represents the average densitometric value from three experiments, normalized against that of GAPDH. (B) U251 glioblastoma multiforme GBM cells were co-immunostained with FTL antibodies (green) and GADD45A antibodies (red), and nuclei were stained with DAPI (blue). (C) Co-immunoprecipitation and immunoblotting analyses confirmed the interaction between FTL and GADD45A proteins. (D) Results of a Cell Counting Kit-8 (CCK8) assay where U251 cells were transfected with the indicated plasmids for 48 h. Cell viability, relative to cells transfected with a control plasmid (vector), is indicated. (E) GADD45A protein levels in FTL expression vector-transfected 293T cell lysates as measured by immunoblotting. Along with elevated FTL protein levels, a decrease in GADD45A protein levels was observed; this effect was attenuated by treatment with a proteasome inhibitor, MG132. The number at the top of each band represents the average densitometric value from three experiments, normalized against that of GAPDH.</p

Subcellular localization of ferritin light chain in glioblastoma multiforme cell lines during cell cycle phases.

<p>(A–C) U251, A172, and U87 glioblastoma multiforme cells in different phases of the cell cycle were immunostained with ferritin light chain (FTL) antibodies (green). Nuclei were stained with DAPI (blue).</p

Robust Succinonitrile-Based Gel Polymer Electrolyte for Lithium-Ion Batteries Withstanding Mechanical Folding and High Temperature

Fabrication of a gel polymer electrolyte containing succinonitrile (GPE-SN) with high mechanical strength is quite challenging because the SN electrolyte always suppresses the formation of polymer networks during in situ polymerization. In this work, a mechanically robust GPE-SN was successfully prepared by using a solution immersion method. During fabrication, the paste-like SN electrolyte was transformed into a liquid SN electrolyte with low viscosity by heating at 50 °C and then infiltrated into the UV-cured highly cross-linked polyurethane acrylate (PUA) skeleton. The resulted GPE-SN film exhibits superior tensile strength (6.5 MPa) compared to the one (0.5 MPa) prepared by in situ polymerization (GPE-SN-IN). The high mechanical strength of the GPE-SN-IM film enables the LiCoO₂/Li₄Ti₅O₁₂ film battery to withstand 100-cycle folding without electrolyte damage and capacity loss. Besides, the GPE-SN presents a high ionic conductivity (1.63 × 10^–3 S·cm^–1 at 25 °C), which is comparable to GPE with a commercial liquid electrolyte (GPE-LE). Because of good thermal stability of the GPE-SN, the LiCoO₂/Li cell with this electrolyte shows better charge–discharge cycling stability than that with GPE-LE at high temperature (55 °C). Thus, the GPE-SN prepared by our method could be a promising polymer electrolyte offering better safety and reliability for lithium-ion batteries

Adsorption effect of Fe₃O₄@Au-C225 to antibody Fc.

<p>Adsorption effect of Fe₃O₄@Au-C225 to antibody Fc.</p

Preparation and characterization of Fe₃O₄@Au-C225 composite targeted nanoparticles for MRI of human glioma - Fig 3

<p>The MRI T2*WI of the human transplanted glioma in mice labeled by Fe3O4@Au-C225 composite targeted MNPs(a), Fe3O4@Au composite MNPs(b), both Fe3O4@Au-C225 composite targeted MNPs and C225(c) (circle for the glioma)(A); The T2*WI relaxation time of the human glioma transplanted in nude mice labeled by Fe3O4@Au-C225 composite targeted MNPs(a), Fe3O4@Au composite MNPs(b), both Fe3O4@Au-C225 composite targeted MNPs and C225(c) at different time points (*compared with the 0h time point, P<0.05)(B).</p

DataSheet_1_MS4A6A is a new prognostic biomarker produced by macrophages in glioma patients.xlsx

MS4A6A has been recognized as being associated with aging and the onset of neurodegenerative disease. However, the mechanisms of MS4A6A in glioma biology and prognosis are ill-defined. Here, we show that MS4A6A is upregulated in glioma tissues, resulting in unfavorable clinical outcomes and poor responses to adjuvant chemotherapy. Multivariate Cox regression analysis suggested that MS4A6A expression can act as a strong and independent predictor for glioma outcomes (CGGA1: HR: 1.765, p < 0.001; CGGA2: HR: 2.626, p < 0.001; TCGA: HR: 1.415, p < 0.001; Rembrandt: HR: 1.809, p < 0.001; Gravendeel: HR: 1.613, p < 0.001). A protein–protein interaction (PPI) network revealed that MS4A6A might be coexpressed with CD68, CD163, and macrophage-specific signatures. Enrichment analysis showed the innate immune response and inflammatory response to be markedly enriched in the high MS4A6A expression group. Additionally, single-cell RNA sequencing (scRNA-seq) analysis revealed distinctive expression features for MS4A6A in macrophages in the glioma immune microenvironment (GIME). Immunofluorescence staining confirmed colocalization of CD68/MS4A6A and CD163/MS4A6A in macrophages. Correlation analysis revealed that MS4A6A expression is positively related to the tumor mutation burden (TMB) of glioma, displaying the high potential of applying MS4A6A to evaluate responsiveness to immunotherapy. Altogether, our research indicates that MS4A6A upregulation may be used as a promising and effective indicator for adjuvant therapy and prognosis assessment.</p