HDAC3 Contributes to Ischemic Stroke by Regulating Interferon Pathway.

BACKGROUND: The inflammation and immune response contribute to ischemic stroke pathology. Damaged brain cells release inflammatory substances to activate the immune system in the acute phase of stroke, including altering the interferon signaling pathway. However, the involvement of histone deacetylation in stroke remains unclear. METHODS: To investigate whether histone deacetylation modulation could regulate the interferon signaling pathway and mediate the pathogenic changes after stroke, the middle cerebral artery occlusion (MCAO) mouse model was treated with histone deacetylase 3 (HDAC3) inhibitor and RGFP966. Additionally, a series of approaches, including middle cerebral artery occlusion (MCAO), real-time polymerase chain reaction (PCR), western blot, 2,3,5-triphenyltetrazolium chloride (TTC) staining, behavioral experiments, and confocal imaging were utilized. RESULTS: It is observed that RGFP966 pretreatment could lead to better outcomes in the MCAO mouse model, including the decrease of infarction volumes, the amelioration of post-stroke anxiety-like behavior, and the relief of inflammatory responses. Furthermore, we found that RGFP966 could counteract the hyperactivation of the interferon signaling pathway and the excessive expression of Z-DNA Binding Protein 1 (ZBP1) in microglia. CONCLUSIONS: We demonstrated a novel mechanism that HDAC3 inhibition could ameliorate the pathological injury after ischemic stroke by downregulating the ZBP1/phosphorylated Interferon Regulatory Factor 3 (p-IRF3) pathway. Thus, these data provide a new promising target for therapies for ischemic stroke.

Ketogenic ß-hydroxybutyrate regulates ß-hydroxybutyrylation of TCA cycle-associated enzymes and attenuates disease-associated pathologies in Alzheimer's mice.

Lysine ß-hydroxybutyrylation (Kbhb) is a post-translational modification that has recently been found to regulate protein functions. However, whether and how protein Kbhb modification participates in Alzheimer's disease (AD) remains unknown. Herein, we carried out 4D label-free ß-hydroxybutylation quantitative proteomics using brain samples of 8-month-old and 2-month-old APP/PS1 AD model mice and wild-type (WT) controls. We identified a series of tricarboxylic acid (TCA) cycle-associated enzymes including citrate synthase (CS) and succinate-CoA ligase subunit alpha (SUCLG1), whose Kbhb modifications were decreased in APP/PS1 mice at pathological stages. Sodium ß-hydroxybutyrate (Na-ß-OHB) treatment markedly increased Kbhb modifications of CS and SUCLG1 and their enzymatic activities, leading to elevated ATP production. We further found that Kbhb modifications at lysine 393 site in CS and at lysine 81 site in SUCLG1 were crucial for their enzymatic activities. Finally, we found that ß-OHB levels were decreased in the brain of APP/PS1 mice at pathological stages. While ketogenic diet not only significantly increased ß-OHB levels, Kbhb modifications and enzymatic activities of CS and SUCLG1, and ATP production, but also dramatically attenuated ß-amyloid plaque pathologies and microgliosis in APP/PS1 mice. Together, our findings indicate the importance of protein Kbhb modification for maintaining normal TCA cycle and ATP production and provide a novel molecular mechanism underlying the beneficial effects of ketogenic diet on energy metabolism and AD intervention.

The combination of temozolomide and perifosine synergistically inhibit glioblastoma by impeding DNA repair and inducing apoptosis.

Temozolomide (TMZ) is widely utilized as the primary chemotherapeutic intervention for glioblastoma. However, the clinical use of TMZ is limited by its various side effects and resistance to chemotherapy. The present study revealed the synergistic inhibition of glioblastoma through the combined administration of TMZ and perifosine. This combination therapy markedly diminished BRCA1 expression, resulting in the suppression of DNA repair mechanisms. Furthermore, the combination of TMZ and perifosine elicited caspase-dependent apoptosis, decreasing glioblastoma cell viability and proliferation. The observed synergistic effect of this combination therapy on glioblastoma was validated in vivo, as evidenced by the substantial reduction in glioblastoma xenograft growth following combined treatment with TMZ and perifosine. In recurrent glioma patients, higher BRCA1 expression is associated with worse prognosis, especially the ones that received TMZ-treated. These findings underscore the potent antitumor activity of the AKT inhibitor perifosine when combined with TMZ and suggest that this approach is a promising strategy for clinical glioblastoma treatment.

MicroRNA expression profile and TNM staging system predict survival in patients with lung adenocarcinoma.

ObjectThe current study was performed to construct a model with microRNA (miRNA/miR) expression profile and TNM staging system for prognosis predicting in patients with lung adenocarcinoma (LUAD). MethodsDifferentially expressed miRNAs were identified from miRNA data of LUAD in The Cancer Genome Atlas (TCGA) database. Potential prognostic miRNAs and TNM classification parameters, screened out by Cox proportional hazards regression analysis, were included in the prognostic model. The prognostic model was visualized with a nomogram, and tested by calculating the C-index and drawing the calibration curve in the training set and validating set, respectively. Finally, the prognostic miRNAs were analyzed with bioinformatics tools. ResultsA total of 194 differentially expressed miRNAs were identified between LUAD tissues and matched normal tissues, including 99 up-regulated and 95 down-regulated miRNAs. miRNA index (miR.index), constructed with nine miRNAs (hsa-let-7i, hsa-mir-1976, hsa-mir-199a-1, hsa-mir-31, hsa-mir-3940, hsa-mir-450a-2, hsa-mir-4677, hsa-mir-548v and hsa-mir-6803), was an independent prognostic indicator for the survival of patients with LUAD. Bioinformatics analysis suggests that the selected miRNAs are involved in the development and progress of LUAD. ConclusionThe prognostic model constructed with nine miRNA expression profile and TNM classification parameters can predict the survival in patients with LUAD, and the predictive power of the model are warranted for further validations.

HER4 promotes the progression of colorectal cancer by promoting epithelial­mesenchymal transition.

Colorectal cancer (CRC) remains one of the most common cancer types worldwide. A few previous studies have examined whether HER4 may promote the progression of CRC. The present study examined the associations among the expression levels of members of the HER family, and investigated the potential mechanism underlying the function of HER4 in CRC cells. Immunohistochemistry analysis was conducted to detect the expression levels of HER family members in patients with CRC. HER4 expression was knocked down using short hairpin RNA in HCT116 cells, and confirmed by quantitative PCR and western blotting. The proliferation and adhesion of CRC cells were analyzed by CCK­8 assays and adhesive assays, respectively. Flow cytometry was used to measure cell apoptosis. Western blotting and immunofluorescence staining in CRC cells were performed to identify proteins related to epithelial­mesenchymal transition. The proportion of patients with CRC presenting positive expression of the HER family members epidermal growth factor receptor (EGFR), HER2, HER3 and HER4 were 72.1, 45.2, 43.8 and 34.2%, respectively. No relationship was found between HER4 and EGFR, HER2 or HER3 expression. Higher expression of HER4 was positively associated with lymph node metastasis (P=0.039). In the present study, HER4 expression was found to be associated with an unfavorable clinical outcome in patients with CRC (Plogrank=0.020). Cell proliferation was inhibited, and apoptosis was increased following HER4 knockdown. Furthermore, HER4 knockdown increased the expression of E­cadherin and decreased the expressions of N­cadherin and vimentin (P<0.05). HER4 expression was found to be unrelated to other HER family members. In the present study, positive expression of HER4 promoted the progression of CRC through epithelial­mesenchymal transition.

Prognostic Impact of PD-1 and Tim-3 Expression in Tumor Tissue in Stage I-III Colorectal Cancer.

BACKGROUND: Programmed cell death receptor 1 (PD-1) and T cell immunoglobulin mucin-3 (Tim-3) are considered as important immunosuppressive molecules and play an important role in tumor immune escape and cancer progression. However, it remains unclear whether PD-1 and Tim-3 are coexpressed in stage I-III colorectal cancer (CRC) and how they impact on the prognosis of the disease. MATERIALS AND METHODS: A total of two cohorts with 451 patients who underwent surgery for stage I-III CRC treatment were enrolled in the study. Among which, 378 cases were from The Cancer Genome Atlas (TCGA) database and 73 cases were from the Fourth Hospital of Hebei Medical University (FHHMU) cohort. The mRNA expressions of PD-1 and Tim-3 in tumor tissue in stage I-III CRC were obtained from TCGA database. Immunohistochemistry was used to assess the expressions of PD-1 and Tim-3 in tumor tissue in stage I-III CRC in the FHHMU cohort. Interactive relationships between PD-1 and Tim-3 were retrieved through the online STRING database, which was used to study the interactions between proteins. DAVID, consisting of comprehensive biological function annotation information, was applied for the GO and KEGG pathway enrichment analysis of the interactive genes. RESULTS: In the FHHMU cohort, the high expressions of PD-1 and Tim-3 were, respectively, found in 42.47% and 84.93% of stage I-III CRC tissue. PD-1 was significantly associated with age, primary site, and lymphatic metastasis. Tim-3 was closely related to the primary site. Correlation analysis showed that PD-1 and Tim-3 were positively correlated (r = 0.5682, P < 0.001). In TCGA cohort, PD-1 and Tim-3 were associated with the prognosis of CRC patients in terms of 5-year survival (P < 0.05). In the FHHMU cohort, the 5-year survival of patients with high levels of PD-1 and Tim-3 was 54.84% and 65.85%, respectively. Among which, the high expression of PD-1 was associated with poor prognosis (5-year OS: 54.84% vs. 88.10%, P = 0.003). The 5-year survival rate of CRC patients with coexpression of PD-1 and Tim-3 was 45.00%, which was significantly worse than non-coexpression (72.73%, 85.71%, and 90.48% separately). The functional network of PD-1 and Tim-3 primarily participates in the regulation of immune cell activation and proliferation, immune cell receptor complex, cell adhesion molecules, and T cell receptor signaling pathway. CONCLUSION: In summary, upregulation of PD-1 and Tim-3 in stage I-III CRC tumor tissue could be associated with the poor prognosis of patients. Those patients with coexpression of PD-1 and Tim-3 may have a significantly worse prognosis.

HER4 isoform CYT2 and its ligand NRG1III are expressed at high levels in human colorectal cancer.

The present study aimed to evaluate the expression of human epidermal growth factor receptor (HER4) isoforms and their ligand neuregulin 1 (NRG1) isoforms in human primary colorectal cancer (CRC). The mRNA expression of HER4 isoforms JM-a, JM-b, CYT1 and CYT2, and their ligand isoforms NRG1 I, II and III in CRC tissues and adjacent normal tissues were quantified by reverse transcription-quantitative polymerase chain reaction analysis. Univariate analysis and logistic regression analysis were performed to analyze the association between HER4 and NRG1 expression and lymph node metastasis in CRC. The expression levels of CYT1 (P=0.002), CYT2 (P=0.002) and NRG1 type III (P<0.001) were significantly higher in the CRC tissues than in the adjacent normal tissues. The expression of CYT2 was correlated with tumor stage (P=0.018), lymph node status (P=0.015) and tumor-node-metastasis (P=0.038) in CRC. The expression of NRG1III was correlated with lymph node metastasis, and the expression of CYT2 was associated with the expression of NRG1III (r=0.691, P<0.01). The logistic regression analysis indicated that expression of CYT2 >50 was a risk factor for lymph node metastasis in CRC. In conclusion the expression levels of CYT1, CYT2 and NRG1III were upregulated in CRC. An expression of CYT-2 >50 was identified as a risk factor for lymph node metastasis in CRC. Therefore, CY-2 and NRG1III may be involved in the progression of CRC.