Predominant Expression Research Articles

The Mining for Flowering-Related Genes Based on De Novo Transcriptome Sequencing in the Endangered Plant Phoebe chekiangensis

Phoebe chekiangensis is an indigenous, endangered, and valuable timber and garden tree species in China, which is notable for having a short juvenile phase (early flowering), unique among the Phoebe genus. However, the molecular mechanisms regulating the flowering of P. chekiangensis remain unexplored, primarily due to the lack of transcriptomic or genomic data. In the present study, transcriptome sequencing yielded 53 million RNA reads, resulting in 111,250 unigenes after de novo assembly. Of these, 47,525 unigenes (42.72%) were successfully annotated in the non-redundant (Nr) database. Furthermore, 15,605 unigenes were assigned to Clusters of Orthologous Groups (KOGs), and 36,370 unigenes were classified into Gene Ontology (GO) categories. A total of 16,135 unigenes were mapped to the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, involving 298 pathways. Based on the expression levels, Gibberellin signaling pathway-related genes were the most predominant expression levels. Hormonal analysis showed that gibberellin (GA) levels varied across tissues and flowering stages, as GA20 levels in leaves were low during full bloom, while GA1 and GA5 levels peaked in flowers. Furthermore, several key genes involved in gibberellin biosynthesis, including CPS, GID1, GA20ox, GA3ox, and GA2ox, exhibited stage-specific expression patterns. Certain genes were highly expressed during the initial phases of flowering, while others, like GA3ox and GA2ox, reached peak expression at full bloom. These findings provide valuable insights into the molecular mechanisms underlying flowering in P. chekiangensis, laying the foundation for future breeding efforts. This transcriptome dataset will serve as an important public resource for molecular research on this species, facilitating the discovery of functional genes related to its growth, development, and flowering regulation.

CD271high cancer stem cells regulate macrophage polarization in head and neck squamous cell carcinoma.

Cancer stem cells (CSCs) are considered key drivers of progression in head and neck squamous cell carcinoma (HNSCC). Our single-cell RNA sequencing (scRNA-seq) analysis revealed predominant expression of CD271 in CSCs, however, its role as a CSC marker in HNSCC requires further elucidation. We investigated the stemness characteristics of CD271high HNSCC cells and their interactions with the tumor immune microenvironment. scRNA-seq data from hypopharyngeal squamous cell carcinoma (HPSCC) tissues were analyzed to identify expression profile of CSCs. Overall survival was compared between CD271high and CD271low patients based on immunostaining of HPSCC samples. The stemness of CD271high HNSCC cells was evaluated via an in vivo limiting dilution assay. In a C57BL/6 mice model, the percentage of immune cells and macrophage subtypes were analyzed by flow cytometry. The role of CD271 in macrophage polarization was further examined by in vitro coculture of CD271high cells with CD14+ monocytes. Gene expressions were analyzed by qPCR. CD271 is predominantly expressed in CSCs identified by scRNA-seq analysis. CD271 enhances HNSCC cell proliferation and is negatively correlated with patient prognosis in HPSCC. CD271 knockdown suppressed HNSCC tumor growth and regulated macrophage polarization within the TME. CD271high cells exhibited stemness features and enhanced tumor growth in vivo. CD271high HNSCC cells exhibit CSC characteristics and regulate macrophage polarization. Targeting CD271 may improve the immunosuppressive TME to inhibit tumor growth. Combining CD271-targeting agents with other therapies presents a promising strategy that may enhance therapeutic efficacy and prognosis in HNSCC.

Neuroanatomical and functional substrates of the short video addiction and its association with brain transcriptomic and cellular architecture.

Short video addiction (SVA) has emerged as a growing behavioral and social issue, driven by the widespread use of digital platforms that provide highly engaging, personalized, and brief video content. We investigated the neuroanatomical and functional substrates of SVA symptoms, alongside brain transcriptomic and cellular characteristics, using Inter-Subject Representational Similarity Analysis (IS-RSA) and transcriptomic approaches. Behaviorally, we found that dispositional envy was associated with SVA. Structurally, SVA was positively correlated with increased morphological volumes in the orbitofrontal cortex (OFC) and bilateral cerebellum. Functionally, the dorsolateral prefrontal cortex (DLPFC), posterior cingulate cortex (PCC), cerebellum, and temporal pole (TP) exhibited heightened spontaneous activity, which was positively correlated with SVA severity. Transcriptomic and cellular analyses also showed specific genes linked to gray matter volume (GMV) associated with SVA, with predominant expression in excitatory and inhibitory neurons. These genes showed distinct spatiotemporal expression patterns in the cerebellum during adolescence. This study offers a comprehensive framework integrating structural, functional, and neurochemical evidence to highlight the neural-transcriptomic underpinnings of SVA symptoms in a non-clinical population.

Exploring the comorbidity mechanisms between atherosclerosis and hashimoto's thyroiditis based on microarray and single-cell sequencing analysis.

Atherosclerosis (AS) is a chronic vascular disease characterized by inflammation of the arterial wall and the formation of cholesterol plaques. Hashimoto's thyroiditis (HT) is an autoimmune disorder marked by chronic inflammation and destruction of thyroid tissue. Although previous studies have identified common risk factors between AS and HT, the specific etiology and pathogenic mechanisms underlying these associations remain unclear. We obtained relevant datasets for AS and HT from the Gene Expression Omnibus (GEO). By employing the Limma package, we pinpointed common differentially expressed genes (DEGs) and discerned co-expression modules linked to AS and HT via Weighted Gene Co-expression Network Analysis (WGCNA). We elucidated gene functions and regulatory networks across various biological scenarios through enrichment and pathway analysis using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Core genes were identified using Cytoscape software and further validated with external datasets. We also conducted immune infiltration analysis on these core genes utilizing the CIBERSORT method. Lastly, Single-cell analysis was instrumental in uncovering common diagnostic markers. Based on differential analysis and WGCNA, we identified 119 candidate genes within the cohorts for AS and HT. KEGG and GO enrichment analyses indicate that these genes are significantly involved in antigen processing and presentation, along with various immune-inflammatory pathways. Two pivotal genes, PTPRC and TYROBP, were identified using five algorithms from the cytoHubba plugin. Validation through external datasets confirmed their substantial diagnostic value for AS and HT. Moreover, the results of Gene Set Enrichment Analysis (GSEA) indicated that these core genes are significantly enriched in various receptor interactions and signaling pathways. Immune infiltration analysis revealed a strong association of lymphocytes and macrophages with the pathogenesis of AS and HT. Single-cell analysis demonstrated predominant expression of the core genes in macrophages, monocytes, T cells and Common Myeloid Progenitor (CMP). This study proposes that an aberrant immune response might represent a shared pathogenic mechanism in AS and HT. The genes PTPRC and TYROBP are identified as critical potential biomarkers and therapeutic targets for these comorbid conditions. Furthermore, the core genes and their interactions with immune cells could serve as promising targets for future diagnostic and therapeutic strategies.

Characterization of sulfakinin and its role in larval feeding and molting in Spodoptera frugiperda.

Feeding and molting are particularly important physiological processes for insects, and it has been reported that neuropeptides are involved in the nervous regulation of these 2 processes. Sulfakinin (SK) is an important neuropeptide that is widely distributed among insects and plays a pivotal role in regulating feeding, courtship, aggression, and locomotion. In this study, we investigated the involvement of SK in feeding and molting on a highly notorious pest insect, the fall armyworm, Spodoptera frugiperda. SK transcript levels were found in all larval stages and there was a predominant expression of SK in the brain of 5th instar larvae. By immunostaining, SK was detected in 2 pairs of cells in the median protocerebrum. But during prolonged periods of starvation, there was a significant reduction in SK messenger RNA levels; however, subsequent refeeding led to a notable increase. To investigate the role of SK in feeding and molting, SK was silenced in S. frugiperda larvae through RNA interference. This resulted in a significant increase in food intake, weight gain, and the molting process happened more rapidly in the double-stranded SK-treated larvae compared to the controls. Conversely, injection of sulfated SK peptide (sSK) caused opposite effects. Interestingly, SK-knockdown in larvae resulted in increased levels of 20-hydroxyecdysone and also of the expression of some of it signaling pathway genes. Altogether, this study highlights the important role played by SK in regulating feeding and molting in S. frugiperda.

Identification and Functional Analysis of PANoptosis-Associated Genes in the Progression From Sepsis to ARDS.

Sepsis and acute respiratory distress syndrome (ARDS) are common inflammatory conditions in intensive care, with ARDS significantly increasing mortality in septic patients. PANoptosis, a newly discovered form of programmed cell death involving multiple cell death pathways, plays a critical role in inflammatory diseases. This study aims to elucidate the PANoptosis-related genes (PRGs) and their involvement in the progression of sepsis to ARDS. This study analyzed differentially expressed genes (DEGs) associated with PRGs to explore their role in the progression of immune disorders from sepsis to septic ARDS. A diagnostic prediction model was constructed based on key PRGs identified through bioinformatics analysis. Functional enrichment analyses were conducted to determine pathway involvement, and correlations with immune cells were assessed. Mendelian randomization analysis was applied to investigate potential causal links between specific PRGs and ARDS. Immunohistochemical analysis was used to evaluate PRG expression in lung tissue. The prediction model effectively distinguished septic ARDS patients from those with sepsis. NDRG1 expression was elevated in ARDS, while DDX3X, PTPRC, and TNFSF8 were downregulated. NDRG1 showed a positive correlation with activated dendritic cells, whereas DDX3X, PTPRC, and TNFSF8 were positively associated with neutrophils and negatively correlated with CD56bright NK cells. Functional enrichment analysis highlighted spliceosome function, MAPK signaling, endocytosis, and antigen processing pathways as significantly associated with these PRGs. Mendelian randomization suggested a causal link between NDRG1 and ARDS, and immunohistochemical analysis revealed its predominant expression near vascular walls. In a mouse model of sepsis, suppression of NDRG1 alleviated lung injury. PANoptosis may contribute to immune dysregulation in sepsis-associated ARDS. NDRG1 is identified as a potential therapeutic target, offering new avenues for mitigating ARDS progression and improving patient outcomes.

Crosstalk between CD180-overexpression macrophages and glioma cells worsens patient survival through malignant phenotype promotion and immunosuppressive regulation

BackgroundUnderstanding the molecular mechanisms in immunosuppressive regulation is crucial for improving immunotherapeutic strategies. Macrophages, the major immune cells in tumor microenvironment (TME), play a dual role in tumor progression. CD180, primarily expressed in macrophages, remains unclear and requires further investigation.MethodsRNA-seq data were obtained to analyze CD180 expression in gliomas and assess its prognostic value. The comprehensive immune infiltration analysis was performed. Single-cell RNA-seq (scRNA-seq) data were used to examine CD180 expression distribution at the cellular level. CD180-overexpression macrophages were co-cultured with three glioma cell lines. The effects on glioma cell behavior were evaluated through qRT-PCR, Western blot, CCK-8 assay, EdU assay, Transwell assay, TUNEL assay, and flow cytometry. Differentially expressed genes (DEGs) and their potential biological functions were analyzed between different CD180 expression groups. Consensus clustering was used to identify CD180-related glioma subtypes.ResultsCD180 was significantly upregulated in glioma samples and associated with poor prognosis. High CD180 expression was correlated with increased immune cell infiltration, particularly macrophages, and elevated levels of immune checkpoints. Analysis of scRNA-seq data revealed the predominant expression of CD180 in macrophages within the glioma TME. In vitro experiments demonstrated that CD180-overexpression macrophages promoted glioma cell proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT), while decreasing apoptosis. Mutations in TP53 and PTEN were significantly more prevalent in the high CD180 expression group. We identified nine chemotherapeutic agents that were more effective in glioma patients with high CD180 expression. Additionally, two CD180-related glioma subtypes with distinct prognosis were identified.ConclusionsThis study confirmed the prognostic role of CD180 in glioma and its involvement in immunosuppressive regulation and malignant phenotype promotion. Therefore, CD180 was considered as a promising target for immunotherapeutic strategies in glioma treatment.

CD2AP deficiency aggravates Alzheimer’s disease phenotypes and pathology through p38 MAPK activation

BackgroundAlzheimer’s disease (AD) is the most common form of neurodegenerative disorder, which is characterized by a decline in cognitive abilities. Genome-wide association and clinicopathological studies have demonstrated that the CD2-associated protein (CD2AP) gene is one of the most important genetic risk factors for AD. However, the precise mechanisms by which CD2AP is linked to AD pathogenesis remain unclear.MethodsThe spatiotemporal expression pattern of CD2AP was determined. Then, we generated and characterized an APP/PS1 mouse model with neuron-specific Cd2ap deletion, using immunoblotting, immunofluorescence, enzyme-linked immunosorbent assay, electrophysiology and behavioral tests. Additionally, we established a stable CD2AP-knockdown SH-SY5Y cell line to further elucidate the specific molecular mechanisms by which CD2AP contributes to AD pathogenesis. Finally, the APP/PS1 mice with neuron-specific Cd2ap deletion were treated with an inhibitor targeting the pathway identified above to further validate our findings.ResultsCD2AP is widely expressed in various regions of the mouse brain, with predominant expression in neurons and vascular endothelial cells. In APP/PS1 mice, neuronal knockout of Cd2ap significantly aggravated tau pathology, synaptic impairments and cognitive deficits. Mechanistically, the knockout of Cd2ap activated p38 mitogen-activated protein kinase (MAPK) signaling, which contributed to increased tau phosphorylation, synaptic injury, neuronal apoptosis and cognitive impairment. Furthermore, the phenotypes of neuronal Cd2ap knockout were ameliorated by a p38 MAPK inhibitor.ConclusionOur study presents the first in vivo evidence that CD2AP deficiency exacerbates the phenotypes and pathology of AD through the p38 MAPK pathway, identifying CD2AP/p38 MAPK as promising therapeutic targets for AD.

Genome-Wide Transcriptional Profiling and Functional Analysis Reveal That OsPHT4;4 Is Critical for the Growth and Development of Rice.

Phosphorus (P) is an essential macronutrient required for various vital processes in crop growth and development, including signal transduction, CO2 fixation, and photosynthetic phosphorylation. Phosphate transporters (PHTs) in plants play critical roles in the uptake, distribution, and internal transport of Phosphate (Pi). Among these transporters, the PHT4 family is widely distributed across plant species; however, the specific functions of many members within this family remain to be fully elucidated. This study focuses on unraveling the function of OsPHT4;4 in Pi utilization and photoprotection. The findings demonstrate that OsPHT4;4 acts as a low-affinity Pi transporter localized to the chloroplast membrane and reveal predominant expression of OsPHT4;4 in leaves, with peak expression during tillering and clear induction by light, exhibiting circadian rhythmicity. The ospht4;4 mutants display stunted growth. Transcriptomic analysis comparing ospht4;4 mutants and wild-types (WT) identified 1482 differentially expressed genes (DEGs), including 729 upregulated genes and 753 downregulated genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis reveals enrichment DEGs related to photosynthesis-antenna proteins, carbohydrate metabolism, and phenylpropanoid biosynthesis. These findings suggest that OsPHT4;4 plays crucial roles not only in photosynthesis but also in plant defense as an integral component involved in Pi metabolism.

Unlocking the role of miR-17: Driving G1-S cell cycle transition in oral tongue cancer through integrated bioinformatics and laboratory analyses.

This study aims to identify miRNA-mediated regulation of the cell cycle in oral tongue cancer. Comprehensive computational analysis was performed on the GEO dataset "GSE168227". DIANA Tool-mir path v.3, STRING, Cytoscape 3.6.0, Enrichr, and TargetScan Human 7.2 were utilized to identify and analyze miRNAs and their targets in oral tongue cancer. The identified miRNA and its target genes were further analyzed in oral tongue cancer patients using qPCR and immunohistochemistry (IHC). Computational analysis revealed miR-17 as a differentially expressed miRNA in oral tongue cancer. Database analysis indicated potential binding sites of miR-17 for CDKN1A and CCND1 mRNA at 3'-UTR. In oral tongue cancer samples, miR-17, CDKN1A, and CCND1expression were upregulated compared to controls. IHC demonstrated overexpression of p21 and Cyclin D1 across various tumor grades, with predominant cytoplasmic expression of p21 observed in oral tongue cancer samples. The findings suggest that miR-17 may regulate the G1-S transition of the cell cycle in oral tongue cancer. Further validation and functional studies are warranted to confirm their role as biomarkers.

ZmICE1a regulates the defence-storage trade-off in maize endosperm.

The endosperm of cereal grains feeds the entire world as a major food supply; however, little is known about its defence response during endosperm development. The Inducer of CBF Expression 1 (ICE1) is a well-known regulator of cold tolerance in plants. ICE1 has a monocot-specific homologue that is preferentially expressed in cereal endosperms but with an unclear regulatory function. Here we characterized the function of monocot-specific ZmICE1a, which is expressed in the entire endosperm, with a predominant expression in its peripheral regions, including the aleurone layer, subaleurone layer and basal endosperm transfer layer in maize (Zea mays). Loss of function of ZmICE1a reduced starch content and kernel weight. RNA sequencing and CUT&Tag-seq analyses revealed that ZmICE1a positively regulates genes in starch synthesis while negatively regulating genes in aleurone layer-specific defence and the synthesis of indole-3-acetic acid and jasmonic acid (JA). Exogenous indole-3-acetic acid and JA both induce the expression of numerous defence genes, which show distinct spatial-specific expression in the basal endosperm transfer layer and subaleurone layer, respectively. Moreover, we dissected a JA-ZmJAZ9-ZmICE1a-MPI signalling axis involved in JA-mediated defence regulation. Overall, our study revealed ZmICE1a as a key regulator of endosperm defence response and a coordinator of the defence-storage trade-off in endosperm development.

Concentration and composition of circulating vesicles of adipocytic origin in patients with colon polyps and colorectal cancer

Extracellular vesicles (EVs) are a heterogeneous population of membrane particles less than 1 μm in size, secreted by various cell types. Most EVs circulating in human blood are particles of platelet, leukocyte, erythrocyte and endothelial origin. The composition of circulating adipocyte EVs in various pathological conditions has been virtually unknown. Small EVs from the blood plasma of patients with colorectal cancer (CRC) and colon polyps with obesity or metabolic syndrome were isolated by ultrafiltration with double ultracentrifugation. To study the composition of adipocyte EVs, immunoprecipitation in combination with Western blotting and flow cytometry were used. Vesicle fractions (FABP4- and CD11b-immunoprecipitated EVs, as well as EVs contained in the supernatant after removal of CD11b-positive EVs) contained a complex of adipocyte markers (FABP4, PPAR-γ and perilipin 1). EVs of monocyte-macrophage origin precipitated on CD11b-coated particles in CRC patients without obesity were characterized by combined overexpression of FABP4 and perilipin 1, while such overexpression was not typical for CRC patients with metabolic syndrome or obesity. The fraction of truly adipocyte vesicles (supernatant after removal of CD11b-positive EVs) was characterized by the presence in all patients of a complex of adipocyte markers with predominant expression of FABP4 in both patients with metabolic syndrome/metabolically healthy obesity and patients without metabolic disorders. To correctly characterize circulating EVs of patients without obesity, it is first necessary to remove the fraction of CD11b-positive monocyte-macrophage EVs from EV preparations by immunoprecipitation or similar methods, and in the supernatant after removal/sorption of precipitated EVs, the composition of adipocyte vesicles can be studied using a set of markers (FABP4, PPAR-γ, perilipin 1, etc.). Moreover, in patients with metabolic disorders, taking into account the insignificant expression of FABP4 in CD11b-immunoprecipitated EVs, preliminary depletion of vesicle preparations is apparently not so necessary.

Unveiling the role of Pleckstrin-2 in tumor progression and immune modulation: insights from a comprehensive pan-cancer analysis with focus on lung cancer

Cancer remains a leading cause of mortality globally, highlighting the need for novel biomarkers to enhance prognosis and therapeutic strategies. Pleckstrin-2 (PLEK2), a member of the pleckstrin family, has been implicated in processes critical to tumor progression, but its role across cancers remains underexplored. This study systematically examined the expression patterns, prognostic relevance, and functional impact of PLEK2 across multiple cancer types. Using data from The Cancer Genome Atlas (TCGA), Genotype Tissue Expression Project (GTEx), and the Human Protein Atlas, we analyzed PLEK2 expression in both cancerous and normal tissues, revealing significant overexpression of PLEK2 in various cancers at the mRNA and protein levels. Single-cell RNA sequencing further indicated predominant expression of PLEK2 in tumor cells and macrophages within the tumor microenvironment. Survival analysis demonstrated that elevated PLEK2 expression correlated with poor prognosis in specific cancers, though its impact varied across cancer types. Functional assays showed that PLEK2 knockdown inhibited proliferation and migration in human cancer cell lines. In vivo studies using a Lewis lung carcinoma (LLC) model confirmed that PLEK2 knockdown suppressed tumor growth and enhanced the efficacy of PD-1 immunotherapy. Mechanistically, PLEK2 knockdown was associated with reduced AKT pathway activation, diminished tumor-associated macrophage infiltration, and increased CD8 T cell presence. Compounds like Navitoclax were also identified as potential PLEK2 inhibitors. In conclusion, PLEK2 played a multifaceted role in cancer progression and immune response modulation. Targeting PLEK2 might suppress tumor growth and overcome immunotherapy resistance, offering a promising biomarker and therapeutic target to improve cancer treatment outcomes.

MRNA delivery enabled by metal–organic nanoparticles

mRNA therapeutics are set to revolutionize disease prevention and treatment, inspiring the development of platforms for safe and effective mRNA delivery. However, current mRNA delivery platforms face some challenges, including limited organ tropism for nonvaccine applications and inflammation induced by cationic nanoparticle components. Herein, we address these challenges through a versatile, noncationic nanoparticle platform whereby mRNA is assembled into a poly(ethylene glycol)-polyphenol network stabilized by metal ions. Screening a range of components and relative compositional ratios affords a library of stable, noncationic, and highly biocompatible metal–organic nanoparticles with robust mRNA transfection in vitro and in mice. Intravenous administration of the lead mRNA-containing metal–organic nanoparticles enables predominant protein expression and gene editing in the brain, liver, and kidney, while organ tropism is tuned by varying nanoparticle composition. This study opens an avenue for realizing metal–organic nanoparticle-enabled mRNA delivery, offering a modular approach to assembling mRNA therapeutics for health applications.

Epigenomic Alterations of the Human CYP11B Gene in Adrenal Zonation.

The CYP11B2 gene is sporadically expressed in the zona glomerulosa (ZG), whereas the CYP11B1 gene is detected in the zona fasciculata (ZF)/reticularis (ZR), with predominant expression in the ZF. We studied the association between DNA methylation and adrenal zonation. Next, the CYP11B2 methylation statuses in the adrenal medulla (n = 4) and pheochromocytomas (n = 7) were examined. The expression of CYP11B2 in pheochromocytomas and non-functioning adenomas (NFAs) (n = 4) was also studied. Adrenals from five autopsy subjects were assessed for immunohistochemically defined adrenal zonation. We used laser capture microscopy to isolate DNA from each zone in adrenal tissues. CYP11B1 was predominantly unmethylated in the ZF but heavily methylated in the ZG and the ZR. In contrast, CYP11B2 was hypomethylated in the ZG compared with in the ZF and the ZR. In terms of the expression site and strength, the promoter methylation patterns for CYP11B2 and CYP11B1 showed capacities to express CYP11B enzymes. The DNA methylation patterns of the CYP11B2 and CYP11B1 promoters were closely associated with adrenal zonation. The unmethylated CpGs of CYP11B2 were found in the adrenal medulla and pheochromocytomas. Gene expression of CYP11B2 was detected in the pheochromocytomas. These results indicate the possibility that the synthesis of aldosterone occurs in the adrenal medulla. Further study is necessary to elucidate the pathophysiological roles for the synthesis of aldosterone in the adrenal medulla.

Fusion of breast cancer MCF-7 cells with mesenchymal stem cells rearranges interallelic gene expression and enhances cancer malignancy

Fusion among normal cells is tightly regulated and required for the developmental processes of an organism. Cancer cell fusion appears relatively rare but is associated with generating new hybrid cancer cells with aggressive properties. However, it remains unclear how cancer cells acquire aggressiveness via cell fusion. Here, we report changes in cell proliferative capacity, cell motility, anticancer drug resistance, and gene expression profiles when fusing human MCF-7 breast cancer cells and mesenchymal stem cells (MSCs). The fused cells were established using envelopes of a hemagglutinating virus of Japan, which increased cell proliferation, motility, and drug resistance. Comprehensive gene expression profile analysis revealed that the fused cells expressed higher levels of glycolysis-related genes than their parental cells. In fact, the fused cells relied more on glycolysis for ATP production (Warburg effect). HIF1A, which induces the expression of glycolysis-related genes, was upregulated in fused cells compared to MCF-7 cells. Allele-specific expression analysis of the fused cells indicated that MSC allele-derived HIF1A efficiently induces the expression of glycolysis-related genes in the MCF-7 allele. These findings indicate that the reorganization of gene expression by combining MSCs and MCF-7 alleles resulted in the predominant expression of glycolysis-related genes and increased malignancy in the fused cells.

L-PGDS-PGD2-DP1 Axis Regulates Phagocytosis by CD36+ MGs/MΦs That Are Exclusively Present Within Ischemic Areas After Stroke.

Brain injuries, such as ischemic stroke, cause cell death. Although phagocytosis of cellular debris is mainly performed by microglia/macrophages (MGs/MΦs), excessive accumulation beyond their phagocytic capacities results in waste product buildup, delaying brain cell regeneration. Therefore, it is essential to increase the potential for waste product removal from damaged brains. Lipocalin-type prostaglandin D synthase (L-PGDS) is the primary synthase for prostaglandin D2 (PGD2) and has been reported as a scavenger of waste products. However, the mechanism by which the L-PGDS-PGD2 axis exerts such an effect remains unelucidated. In this study, using a mouse model of ischemic stroke, we found that L-PGDS and its downstream signaling pathway components, including PGD2 and PGD2 receptor DP1 (but not DP2), were significantly upregulated in ischemic areas. Immunohistochemistry revealed the predominant expression of L-PGDS in the leptomeninges of ischemic areas and high expression levels of DP1 in CD36+ MGs/MΦs that were specifically present within ischemic areas. Furthermore, PGD2 treatment promoted the conversion of MGs/MΦs into CD36+ scavenger types and increased phagocytic activities of CD36+ MGs/MΦs. Because CD36+ MGs/MΦs specifically appeared within ischemic areas after stroke, our findings suggest that the L-PGDS-PGD2-DP1 axis plays an important role in brain tissue repair by regulating phagocytic activities of CD36+ MGs/MΦs.

ATP-gated P2x7 receptor is a major channel type at type II auditory nerves and required for hearing sensitivity efferent controlling and noise protection.

Hearing sensitivity and noise protection are mediated and determined by negative feedback of the cochlear efferent system. Type II auditory nerves (ANs) innervate outer hair cells (OHCs) in the cochlea and provide an input to this efferent control. However, little is known about underlying channel information. Here, we report that ATP-gated P2x7 receptor had a predominant expression at type II ANs and the synaptic areas under inner hair cells and OHCs with lateral and medial olivocochlear efferent nerves. Knockout (KO) of P2x7 increased hearing sensitivity with enhanced acoustic startle response (ASR), auditory brainstem response (ABR), and cochlear microphonics (CM) by increasing OHC electromotility, an active cochlear amplifier in mammals. P2x7 KO also increased susceptibility to noise. Middle level noise exposure could impair active cochlear mechanics resulting in permanent hearing loss in P2x7 KO mice. These data demonstrate that P2x7 receptors have a critical role in type II AN function and the cochlear efferent system to control hearing sensitivity; deficiency of P2x7 receptors can impair the cochlear efferent suppression leading to hearing oversensitivity and susceptibility to noise.

Chicken GLUT4 function via enhancing mitochondrial oxidative phosphorylation and inhibiting ribosome pathway in skeletal muscle satellite cells

Glucose Transporter 4 (GLUT4) is a crucial protein facilitating glucose uptake and metabolism across cell membranes in mammals. However, information on GLUT4 in birds has historically been limited. In this study, we investigated the dynamic expression profile of chicken GLUT4 using real-time quantitative PCR (RT-qPCR) and examined its potential effects and mechanisms via GLUT4 overexpression and RNA sequencing (RNA-seq) in chicken primary skeletal muscle satellite cells (CP-SMSCs). Our results demonstrated that chicken GLUT4 is differentially expressed across tissues, with predominant expression in skeletal muscles, and across developmental stages of CP-SMSCs, with notable upregulation during the phases of cell proliferation and early differentiation. Notably, 0.1 μM insulin for 60 min significantly elevated the expression of GLUT4 in CP-SMSCs (P < 0.05). GLUT4 overexpression in CP-SMSCs promoted cell proliferation, as evidenced by Cell Counting Kit-8 (CCK-8) (P < 0.05) and 5-Ethynyl-2'-Deoxyuridine (EDU) assays (P < 0.05), and enhanced glucose consumption following 0.1 μM insulin treatment (P < 0.05). However, it inhibited glucose consumption 12 h after the addition of 5 g/L glucose (P < 0.05). After overexpressing GLUT4, we identified 302 differentially expressed genes (DEGs) in CP-SMSCs, with 134 upregulated and 168 downregulated. These DEGs are primarily enriched in pathways such as oxidative phosphorylation, ribosome, cardiac muscle contraction, ATP metabolic processes, and mitochondrial protein complexes. Specifically, in the enriched oxidative phosphorylation pathway, the upregulated DEGs (12) encode mitochondrial proteins, while the downregulated DEGs (6) are nuclear genome-derived. The ribosomal pathway is predominantly inhibited, accompanying with the downregulation of the translocase of outer mitochondrial membrane 7 (TOMM7)/translocase of inner mitochondrial membrane 8 (TIMM8A) complex responsible for mitochondrial protein transport, and a reduction in 28S (LOC121106978) and 18S (LOC112533601) ribosomal rRNAs. In conclusion, chicken GLUT4 is dynamically modulated during development and acts as an insulin responder that significantly regulates cellular glucose uptake and cell proliferation. This regulation occurs mainly through enhancing the mitochondrial oxidative phosphorylation and inhibiting ribosomal pathway.

Activation of NOD1 on tumor-associated macrophages augments CD8+ T cell-mediated antitumor immunity in hepatocellular carcinoma.

The efficacy of immunotherapy targeting the PD-1/PD-L1 pathway in hepatocellular carcinoma (HCC) is limited. NOD-like receptors (NLRs) comprise a highly evolutionarily conserved family of cytosolic bacterial sensors, yet their impact on antitumor immunity against HCC remains unclear. In this study, we uncovered that NOD1, a well-studied member of NLR family, exhibits predominant expression in tumor-associated macrophages (TAMs) and correlates positively with improved prognosis and responses to anti-PD-1 treatments in patients with HCC. Activation of NOD1 in vivo augments antitumor immunity and enhances the effectiveness of anti-PD-1 therapy. Mechanistically, NOD1 activation resulted in diminished expression of perilipin 5, thereby hindering fatty acid oxidation and inducing free fatty acid accumulation in TAMs. This metabolic alteration promoted membrane localization of the costimulatory molecule OX40L in a lipid modification-dependent manner, thereby activating CD8+ T cells. These findings unveil a previously unrecognized role for NOD1 in fortifying antitumor T cell immunity in HCC, potentially advancing cancer immunotherapy.