Transcriptomic Characterization of Copper-Binding Proteins for Predicting Prognosis in Glioma.

BACKGROUND: Copper and copper-binding proteins are key components of tumor progression as they play important roles in tumor invasion and migration, but their associations in gliomas remain unclear. METHODS: Transcriptomic datasets of glioblastoma, low-grade glioma, and normal brain cortex were derived from the TCGA and GTEX databases. Differentially expressed genes (DEGs) of copper-binding proteins were screened and used to construct a prognostic model based on COX and LASSO regression, which was further validated by the CGGA datasets. The expressions of risk-model genes were selectively confirmed via anatomic feature-based expression analysis and immunohistochemistry. The risk score was stratified by age, gender, WHO grade, IDH1 mutation, MGMT promoter methylation, and 1p/19q codeletion status, and a nomogram was constructed and validated. RESULTS: A total of 21 DEGs of copper-binding proteins were identified and a six-gene risk-score model was constructed, consisting of ANG, F5, IL1A, LOXL1, LOXL2, and STEAP3, which accurately predicted 1-, 3-, and 5-year overall survival rates, with the AUC values of 0.87, 0.88, and 0.82, respectively. The high-risk group had a significantly shorter OS (p < 0.0001) and was associated with old age, wild-type IDH1, a high WHO grade, an unmethylated MGMT promoter, and 1p/19q non-codeletion and had higher levels of immune cell infiltration, cancer-immunity suppressor, and immune checkpoint gene expression as well as a higher TMB. CONCLUSIONS: The model based on the genes of copper-binding proteins could contribute to prognosis prediction and provide potential targets against gliomas.

Assessment of 13 essential and toxic trace elements in tumor and peritumoral brain tissues from human glioblastoma.

Trace elements within the brain are important for proper neurological function, but their imbalance has been rarely investigated in glioblastoma. This study enrolled a total of 14 patients with glioblastoma, and the tumor and peritumoral brain tissues were collected while undergoing surgery. The concentrations of Mg, Ca, Cr, Mn, Fe, Co, Cu, Zn, Se, As, Cd, Tl and Pb were determined using a well-evaluated ICP-MS method. The Cu- and Cd-binding proteomes were further analyzed using the anatomic transcriptional atlas from Ivy GAP. Histological evaluation was based on rubeanic acid staining and immunohistochemistry, respectively. The 13 trace element concentrations were obtained, and the highest were Ca, Mn, Fe, Zn and Cu, ranging from a few to dozens of ug/g. Correlation analysis suggested the existence of two intra-correlated clusters: essential metals (Cu-Ca-Zn-Mg) and heavy metals (Pb-As-Cd-Tl-Co-Cr-Mn). Compared to the tumor samples, significantly higher levels of Cu and Cd were observed in the peritumoral region. Further analysis of the Cu- and Cd-binding proteins from the anatomic view suggested that DBH and NOS1 were obviously increased in the leading edge than the central tumor region. Consistent with the above findings, histological evaluation of Cu and DBH further confirmed more copper and DBH expressions in the peritumoral area compared to the tumor core. Trace elements differ in tumor and peritumoral brain zone in glioblastoma, which may associate with tumor angiogenesis.

DIA-MS Based Proteomics Combined with RNA-Seq Data to Unveil the Mitochondrial Dysfunction in Human Glioblastoma.

Mitochondrial dysfunctions underlie the pathogenesis in glioblastoma multiforme (GBM). Comprehensive proteomic profiling of mitochondria-specific changes in human GBM is still insufficient. This study carried out a DIA-MS based proteomic analysis on the mitochondria isolated from human primary GBM and peritumoral tissue (as paired control), and further compared those findings with the transcriptomic datasets. A total of 538 mitochondrion-specific proteins were rigorously confirmed, among which 190 differentially expressed proteins were identified. Co-regulations of the mitochondrial dysfunction pathway networks were observed, including significant up-regulations of mitochondrial translation and apoptosis, as well as down-regulations of OXPHOS and mitochondrial dynamics. Proteins related to FA, AA metabolism and ROS also showed significant variations. Most of these alterations were consistent in trend when compared the proteomics findings with the RNA-Seq datasets, while the changes at protein levels appeared to be more dramatic. Potentially key proteins in GBM were identified, including up-regulated pro-apoptotic protein CASP3, BAX, fatty acid oxidation enzymes CPT1A, CPT2, ACADM, serine-glycine enzymes SHMT2, GATM, ROS-related protein SOD2, GPX1, and CAT; and down-regulated dynamin-related protein MFN1, MFN2, OPA1, and OXPHOS components; and also several differentially expressed ALDH isoforms. This study systematically profiled the mitochondrial dysfunctions by combining proteomic findings and mRNA datasets, which would be a valuable resource to the community for further thorough analyses.

[Identification and management of intra-operative suspicious tissues in 20 transsphenoidal surgery cases].

OBJECTIVE: To determine appropriate protocols for the identification and management of intra operative suspicious tissues during transsphenoidal surgery. METHODS: Clinical data and pathological reports of 20 patients with intra-operative suspicious tissues during transsphenoidal surgeries were analyzed retrospectively. The methods for discriminating between adenoma and normal pituitary tissues were reviewed. RESULTS: The postoperative pathological reports revealed that adenoma and normal pituitary tissues coexisted in 9 samples, while 5 samples were identified as normal pituitary tissues, 2 as adenoma tissues, and 4 as other tissues. Adenomas were distinguished from normal pituitary tissues on the basis of intra-operative appearance, texture, blood supply and possible existence of boundary. CONCLUSION: If decisions are difficult to made during surgeries from the appearance of the suspicious tissues, pathological examinations are advised as a guidance for the next steps.

[Expression and significance of trefoil factor 1 protein and serum pepsinogen in benign and malignant gastric ulcers].

OBJECTIVE: To explore the clinical significance of trefoil factor 1 (TFF1) protein expression and serum pepsinogen (PG) concentration in benign and malignant gastric ulcers. METHODS: The TFF1 protein expression was evaluated by immunohistochemistry in biopsies of gastric mucosa from 18 normal controls, 25 patients with gastric ulcer and 13 patients with ulcerative gastric cancer at our hospital during January to June 2011. The serum concentrations of PGI and PGII were detected by enzyme-linked immunosorbent assay (ELISA) and PG/PGII (PGR) was subsequently calculated. RESULTS: The expression of TFF1 protein increased significantly in ulcerative and peripheral gastric mucosa and peripheral mucosa of gastric cancers versus that in normal controls and the ulcerocancer group (3.04% ± 0.20%, 3.00% ± 0.20%, 3.23% ± 0.26% vs 1.67% ± 0.18%, 0.46% ± 0.18%, all P < 0.01). The elevated expression of TFF1 increased the risk of gastric ulcer (OR: 1.365, 95%CI: 1.065 - 1.749, P = 0.014) while the down-regulation of TFF1 significantly increased the risk of ulcerocancer (OR: 3.067, 95%CI: 1.391 - 6.757, P = 0.005). The serum levels of PGI and PGII in gastric ulcer group were significantly higher than that in normal control and ulcerocancer group ((150 ± 27), (28 ± 9) vs (121 ± 22), (17 ± 7), (79 ± 12), (20 ± 5) µg/L,all P < 0.01). The PGI level and PGR decreased significantly in the ulcerocancer group versus normal control (both P < 0.01). But there was no statistical difference in PGII (P > 0.05). Receiver operating characteristic curve analysis revealed that PGI and PGR were valuable for the diagnosis of malignant gastric cancer with an area under curve of 0.975 and 0.914 respectively. CONCLUSIONS: The expression of TFF1 protein increases in gastric ulcer but decreases in gastric ulcerocancer. The elevated serum levels of PGI and PGII indicate benign ulcer while a marked decline of serum PG I and PGR serves as a risk signal of malignant gastric ulcer. The evaluation of expression profiles of TFF1 protein and PGs is helpful for the differentiation of benign gastric ulcer from malignant ulcerocancer.