Cell Biochemistry and Function最新文献

Metabolic reprogramming plays an essential role in the initiation and aggressiveness of malignant tumors. This study aims to establish a prognostic signature for Wilms tumor (WT) based on metabolism-related genes (MRGs) and to explore potential molecular mechanisms. RNA sequencing data of WT samples and MRGs were sourced from GEO, TCGA and KEGG, respectively. Prognosis-associated differentially expressed MRGs (DE-MRGs) and Lasso regression were employed to create a nine-gene prognostic signature. Internal validation was conducted through bootstrap resampling. Prognostic performance was assessed through Kaplan–Meier curves, ROC curves, the C-index, and calibration curves. Additional analyses encompassed signaling pathways and chemotherapy response prediction. The expression levels and biological functions of NNMT were experimentally validated. A nine-gene signature comprising B3GAT2, COLGALT2, CYP27C1, GAD2, GSTM5, LPIN3, NNMT, ST6GALNAC1 and TCIRG1 was established. The risk score derived from this signature was shown to be an independent prognostic predictor and significantly associated with immune function and autophagy. NNMT expression levels were validated in both cells and tissues. Further experiments in vivo and in vitro indicated that NNMT might influence cholesterol efflux via PPARG-LXRα pathway, thereby enhancing cell proliferation and migration. This study established a metabolism-related gene signature to predict the prognosis of patients with WT. The findings may provide a promising tool for personalized diagnosis and treatment.

The continuous expression of the CRISPR/Cas system in organisms can lead to various potential issues. Some anti-CRISPR strategies have been developed to achieve precise control over CRISPR/Cas, yet these strategies are predominantly protein-based, with the most commonly used anti-CRISPR (Acr) proteins lacking sufficient target specificity. However, in this study, we designed a single-stranded DNA (ssDNA) inhibitor that was complementary to the spacer region on the guide RNA, operating at the nucleic acid level. We demonstrated that this method effectively inhibits the cleavage activity of Cas9-sgRNA ribonucleoprotein (RNP) in a target-specific manner through in vitro cleavage experiments. Furthermore, we explored the binding position and effective length of this inhibitory ssDNA, finding that its inhibitory effect was significantly reduced when the length of continuous complementarity with the 5′ end of the spacer was less than 7nt. The truncated ssDNA also showed potential in reducing off-target effects. Moreover, we applied nucleic acid inhibitors to embryos via microinjection, and gene editing activity was significantly reduced, as evidenced by a decrease in the mosaicism rate of mouse embryos undergoing normal gene editing from (84.4 ± 4.4) % to 0%. Our study introduces a convenient and target-specific nucleic acid inhibitor capable of achieving precise regulation of gene editing.

This study examined the effects of different concentrations of roscovitine (ROSC) on cytoplasmic maturation in bovine oocytes and subsequent embryo development. Oocytes were prematured with ROSC at concentrations of 0, 25, 37.5, 50 and 75 µM for 6 h. A significant increase in the number of oocytes at GV stage was observed in the treatment groups compared to the control group, indicating an inhibitory effect of ROSC on meiosis. After continued culture in a maturation solution without ROSC for 18 h, oocytes did not exhibit significant difference in maturation rates between the treatment and control groups, suggesting that the inhibitory effect of ROSC is reversible. Further analysis revealed that a 37.5 µM ROSC treatment significantly enhanced homogeneous mitochondrial distribution, mitochondrial membrane potential, and glutathione levels in mature oocytes, while reducing reactive oxygen radical content. IVF blastocysts from oocytes treated with 37.5 μM ROSC had a significant increase in the total cell number, and inner cell mass/total cell number ratio, and a reduction in apoptosis rate, despite no significant differences in cleavage or blastocyst rates compared to controls. These findings suggested that prematuration with 37.5 µM ROSC promoted cytoplasmic maturation in bovine oocytes and improved embryo quality. This study analyzed comprehensively the impacts of meiosis arrest induced by ROSC on cytoplasmic maturation in oocyte and embryo quality from various perspectives for the first time.

Metabolic cells exhibit low-grade chronic inflsammation characterized by excessive production and secretion of proinflammatory cytokines and chemokines in response to overnutrition and energy excess. Mitochondrial dysfunction is closely associated with metabolic inflammation. PINK1 (phosphatase and tensin homology-induced putative kinase 1) is a crucial pathway controlling mitochondrial autophagy, essential for maintaining mitochondrial quality control and metabolic homeostasis. The aim of this study was to investigate the role of PINK1 in metabolic inflammation. Our findings indicate that in adipocytes, palmitic acid (PA) activates the expression of PINK1. Additionally, knockdown of PINK1 exacerbates PA-induced adipocyte inflammation. Mechanistically, PINK1 deficiency impairs mitochondrial function, leading to the release of mtDNA and further activation of the cGAS-STING pathway. Therefore, targeting mitochondrial autophagy in adipocytes and the cGAS-STING pathway may represent effective approaches to alleviate the chronic inflammation associated with obesity and related metabolic disorders.

Osteoarthritis is the most prevalent degenerative joint disease, characterized by the trauma and inflammation of the cartilage and subchondral bone, leading to cartilage degradation, reduced bone mass, joint pain, and functional impairment. Articular cartilage and subchondral bone is a complex functional structural complex, in which Netrin protein family and its transmembrane protein receptors such as Unc5b, DCC and A2B are widely expressed. The Netrin family, part of the laminin superfamily, includes various subtypes, with Netrin1 being notably researched for its regulatory role in bone tissue.Netrin1 protein can activate downstream signaling pathways that affect cell function and regulate the functional activity of various cells in joint tissue after binding to its receptors. Studies have pointed out that anti-Netrin1 treatment can significantly alleviate osteoarthritis and other arthritis lesions, as well as reduce neuropathic pain. This review aims to explore the function of the Netrin1 protein and its receptors across joint structures, discussing their potential regulatory mechanisms and implications for treating diseases like osteoarthritis.

M1 macrophages play an important role in pathogenic microorganism infection. Natto, a traditional fermented food from Japan, has long been thought to prevent infection by pathogenic microorganisms. Bacillus subtilis BN strain, which has been isolated from natto, is used in animal feed, health food products, and pharmaceuticals as a probiotic capable of regulating intestinal flora and immunomodulates. The purpose of this study is to investigate the effects of B. subtilis BN strain on the antiviral immune function of human M1 macrophages. The M1 macrophages studied here were derived from a human monocytic cell line (THP-1). Overall, we found that B. subtilis BN strain enhanced gene expressions of antiviral and anti-inflammatory cytokines in M1 macrophages and M1 macrophages stimulated with resiquimod in a cell culture model that mimics single-stranded RNA virus infection. In M1 macrophages, B. subtilis BN strain was also found to promote the gene expression and phosphorylation of key molecules in the toll-like receptor signaling pathways. Furthermore, the BN strain induced the expression of some cytokine mRNA was reduced by the knockdown of NFκB in M1 macrophages. Taken together, these results suggest that toll-like receptor signal pathways are involved in the regulation of antiviral and anti-inflammatory cytokine gene expressions induced by the BN strain in M1 macrophages.

Age-associated B cells (ABCs), an atypical B-cell subset, tend to accumulate with age in mice and humans. These cells exhibit distinct characteristics, such as the ability to secrete antibodies and inflammatory cytokines upon stimulation of Toll-like receptor 7 (TLR7) and TLR9. Additionally, ABCs have been found to be more efficient in presenting antigens to T cells than follicular (FO) B cells. These features contribute to the development of pathogenic phenotypes in aging individuals. In this study, we demonstrated that actin cytoskeleton remodeling was enhanced in CD11b⁺/CD11c⁺ ABCs compared to CD11b⁻/CD11c⁻ B cells. ABCs exhibited higher motility across Transwell membranes and three-dimensional (3D) collagen gels, even without chemoattractants. Due to the remodeling of chemokine receptor expression, ABCs were attracted by CXCL12 and CCL21 rather than CXCL13. Among F-actin remodeling-related factors, expression levels of Fascin1 and Pak1 were increased in ABCs. Treatment with the Pak1 inhibitor, IPA3, significantly attenuated ABC migration in Transwell chambers and 3D collagen gels. In contrast, the Fascin1 inhibitor, migrastatin, only reduced ABC migration in the 3D collagen gel. The increased expression of Fascin1 and Pak1 enhances actin cytoskeleton remodeling in ABCs, facilitating their dispersion within secondary lymphoid tissues.

p53 plays a critical role in the pathogenesis of central nervous system (CNS) diseases, including neurodegenerative diseases, stroke, and neurodevelopmental disorders. Recent studies have shown that p53 regulates pathological processes such as apoptosis, ferroptosis, neuroinflammation, mitochondrial dysfunction, DNA damage, and synaptic dysfunction through complex molecular pathways. Preclinical studies have demonstrated that pharmacological inhibition or knockdown of p53 exerts neuroprotective effects, suggesting its potential therapeutic value. This review systematically summarizes the mechanisms of p53 in CNS diseases and the associated signaling pathways involved in these pathological processes. Additionally, p53-related intervention strategies in CNS diseases and challenges to the development of p53-targeted drugs are summarized, aiming to provide new insights for therapeutic treatment.