Alternative splicing is a crucial mechanism of gene regulation that can be dysregulated in cancer. In pituitary neuroendocrine tumors (PitNETs), alteration in the serine/arginine-rich splicing factors (SRSFs) has been reported. SRSFs phosphorylation and activation is mediated by serine-arginine protein kinase 1 (SRPK1). SRPK1 is considered a proto-oncogene and its inhibition by small molecule inhibitors SRPIN340 and SPHINX31 have shown antitumoral effects via the SRPK1-SRSF1-VEGF pathway modulation in different cancer types. No previous studies have evaluated SRPK1 inhibitors in pituitary tumors. The present work explores the antitumoral effects of SRPIN340 and SPHINX31 in rat and human GH-secreting pituitary tumoral cells. First, immunoblot results showed a reduction of SRSFs phosphorylation induced by both compounds, demonstrating the efficacy of these molecules in inhibiting SRPK1 activity. SRPIN340 reduced GH4C1 cell proliferation (-31.7 (33.6)%, p <0.05 vs control cells at 1µM), cell viability (-16.4 (27.9)%, p<0.05 vs control cells at 1µM), cell migration (-65.0 (46.3)%, p<0.001 vs control cells at 10µM) and induced cell apoptosis (+40.5 (26.6)%, p<0.05 vs control cells at 10µM). Moreover, SRPIN340 significantly decreased both transcript (-56.3 (38.6)%, p<0.01 vs control cells) and protein levels (-33.5 (3.4)%, p<0.05 vs control cells) of the pro-survival VEGF164a isoform. Similar results have been obtained with SPHINX31. Interestingly, cells incubation with the recombinant VEGF164a protein impaired the decrease of cell migration and cell viability mediated by both SRPK1 inhibitors. As for GH-secreting primary cultures from GH-PitNETs, SRPIN340 incubation resulted in reduced VEGF165a expression (-50.6% vs control cells) and GH secretion (-14.45 (8.17)%, p < 0.05 vs control cells). In conclusion, SRPK1 inhibition may represent a novel approach to exert antitumoral effects in somatotroph tumoral cells via SRPK1-SRSF1-VEGF pathway regulation.

Phosphodiesterase-5 (PDE5) inhibitors have been shown to have potential as an adjuvant therapy for cancer. Tadalafil is a potent and selective PDE5 inhibitor that has been shown to inhibit the aggregation of myeloid-derived suppressor cells (MDSC) within tumors. Given the pivotal role of STAT3 signaling in mediating MDSC immunosuppression, we investigated the impact of tadalafil on MDSC differentiation and inhibitory function. Our findings indicate that tadalafil significantly attenuates in situ breast tumor growth and metastasis while impairing the capacity of MDSCs to suppress T-cell proliferation, concomitant with reduced STAT3 phosphorylation. Transcriptomic analysis revealed that tadalafil modulates MDSC metabolism, upregulates NAD+ nucleotidase activity, and disrupts chemotaxis-related transcriptional programs, including downregulation of key chemokine receptors. Consistent with these observations, in vitro migration assays confirmed tadalafil-mediated inhibition of MDSC chemotaxis toward tumor cells. Furthermore, tadalafil markedly enhanced nuclear PARP1 expression, which exhibits a negative regulatory relationship with STAT3. Collectively, these data demonstrate that tadalafil exerts systemic and local immunomodulatory effects on MDSCs during tumor infiltration, primarily through PARP1-STAT3 axis regulation. These insights underscore the therapeutic potential of tadalafil in reprogramming MDSC-mediated immunosuppression in cancer.

Obesity is a metabolic disorder characterized by excessive fat accumulation due to energy imbalance. This study aimed to evaluate the effects of omega-3 fatty acids (n-3 FAs) on body weight, biochemical parameters, thermogenic gene expression, and adipose tissue morphology in a male C57BL/6 mice model of high-fat diet (HFD)-induced obesity initiated at 4 weeks of age. After a one-week adaptation, mice were randomly divided into four groups: a control group fed a standard diet (CD) and three experimental groups fed a HFD alone or enriched with 1.2% or 2.4% n-3 FAs. All groups were fed ad libitum for 12 weeks. Feed intake and body weight were recorded, and glucose and insulin tolerance tests were performed. Serum glucose, lipid profiles, insulin, leptin, and adiponectin levels were analyzed. Gene expression of uncoupling protein 1 (Ucp1), peroxisome proliferator-activated receptor gamma coactivator-1 alpha (Pgc-1α), PR domain containing 16 (Prdm16), and G protein-coupled receptor 120 (Gpr120) was evaluated in brown adipose tissue (BAT), along with histological assessments of both BAT and white adipose tissue (WAT). n-3 FAs improved serum lipid profiles and glucose metabolism without significant changes in body weight or energy intake. Although no significant differences were observed in the expression of Ucp1, Pgc-1α, and Gpr120, Prdm16 expression was significantly higher in the 1.2% HFD group (p < 0.05). Histological analysis showed that n-3 FAs prevented BAT whitening, promoted WAT browning, and reduced inflammatory infiltration. Ucp1 immunoreactivity was significantly lower in the HFD group compared to the CD group (p < 0.05). These findings suggest that n-3 FAs exerts multifaceted protective effects on adipose tissue at genetic, biochemical, and histological levels, highlighting their potential in the management of diet-induced obesity.

Glutamine' synthetase (GS) plays a central role in glutamine metabolism and has been implicated in the progression and treatment resistance of hepatocellular carcinoma (HCC). Although previous studies have explored GS in tumor metabolism, its role in modulating mitophagy and radiosensitivity in HCC cells remains unclear.

Glial cells, including astrocytes, microglia, and oligodendrocytes, play an important role in the repair of damaged central nervous system tissue. In our previous study, we showed that transforming growth factor-beta (TGF-β) signaling occurs in glial cells in the hippocampus after ischemia. However, the functional significance of TGF-β signaling in the hippocampus after ischemia remains unclear. In the present study, transcriptome analysis was performed to comprehensively examine the TGF-β signaling-induced gene expression changes in primary cultured rat mixed glial cells. TGF-β1 upregulated 287 genes and downregulated 272 genes. Representative genes upregulated by TGF-β1 included genes encoding extracellular matrix-related proteins. Conversely, representative genes downregulated by TGF-β1 included genes encoding proteins related to immune response. These results suggest the diverse effects of TGF-β1 on gene expression. Since genes downregulated by TGF-β1 included genes involved in cell phagocytosis, proliferation, and survival, the effects of TGF-β1 and -β2 on cell phagocytosis, proliferation, and survival were investigated in mixed glial cells. TGF-β1 and -β2 suppressed astrocyte and microglial proliferation, and promoted and suppressed astrocyte and microglial phagocytosis, respectively. Additionally, TGF-β1 or -β2 canceled the serum-free culture-induced increase in the ratio of TUNEL-labeled microglia and oligodendrocytes. Furthermore, the culture in a medium containing the TGF-β signaling inhibitor SB525334 reduced glial cell survival and increased the expressions of genes encoding cell death-related molecules. Our study results suggest that TGF-β contributes to postischemic brain tissue repair by regulating glial cell gene expression, phagocytosis, and proliferation, and supporting glial cell survival.

Exogenous plant growth regulators are increasingly applied in viticulture, yet their roles in grape ripening and wine quality remain unclear. In this study, Marselan grapevines were treated with naphthaleneacetic acid (NAA) or gibberellic acid (GA3) at preflowering and preveraison stages, and sugar-acid metabolism and wine sensory traits were evaluated through integrated transcriptomic and metabolomic analyses. Results revealed that NAA applied before flowering upregulated genes for cell expansion and sugar-acid metabolism, increasing alcohols and organic acids, producing wines with high acidity and fresh, fruity aromas. GA3 before veraison enhanced pigment and volatile-aroma pathways, elevating alcohols and esters, resulting in wines with rich floral and ripe-fruit notes and fuller body. Sensory evaluation confirmed that NAA wines were crisp with higher acidity, while GA3 wines had a fuller body and complex aromas. These stage-specific molecular findings provide a framework for precision viticulture and targeted modulation of wine aroma profiles.

Hepatocellular carcinoma (HCC) is a leading cause of cancer-related mortality worldwide, with limited effective therapies and poor prognosis. Although sorafenib and lenvatinib are approved for advanced HCC, their clinical efficacy is often compromised by drug resistance and adverse effects. Bruceine D (BD) has shown antitumor potential in several cancers, but its mechanisms in HCC remain poorly defined.

The cellular and tissue response to acute hypoxia is mediated by a complex adaptive response initiated by hypoxia-inducible factors (HIF) 1 and 2. HIF proteins are key transcription factors that modulate gene expression to adapt cells to the low oxygen tension. Cellular studies of hypoxia have used the low oxygen model by using hypoxia chambers to monitor the gene expression changes, while others have utilized hypoxia mimetics in normoxic conditions. Here, we provide a comparison of the utility and limitations of two hypoxia mimetics by studying the changes in the miRNA and mRNA expression profiles compared to the low oxygen model. We utilized data from NGS-based global analyses, qPCR validation, and HIFa silencing experiments to determine the distinct profiles of HIF-1 and HIF-2 activities of the hypoxia and mimetic models. The ability of CoCl2 and DMOG (dimethyloxalylglycine) to mimic hypoxia was tested in cultured human endothelial cells. The results demonstrated that their treatments resulted in a differential stabilization of HIF-1α and HIF-2α, with CoCl2 predominantly stabilizing HIF-1α, and DMOG stabilizing HIF-2α. While CoCl2 and DMOG show potential in mimicking certain isoform-specific effects, their inability to completely mimic their effects on miRNAs and their mRNA targets reveals that these changes do not fully reproduce the hypoxia-induced effects in pathways related to angiogenesis and apoptosis. Taken together, while the hypoxia mimetics offer valuable insights into the isoform-specific effects of HIF-1 and HIF-2, their inability to fully replicate the complexity of hypoxia-induced miRNA interactions is not illustrated in human endothelial cells.

Mesenchymal stem cells (MSC) play a vital role in bone regeneration. Poly-(l-lactide-co-ε-caprolactone) scaffolds functionalised with modified rhamnogalacturonan-I (RG-I) potato pectin, can modulate inflammation and promote bone regeneration in-vitro and in-vivo by modulating the action of the galactoside-binding galectin-3. In this study, we determined the binding affinity to galectin-3 of potato unmodified RG-I (PU) and its arabinose-deficient form (PA) and investigated the transcriptomic effects of PA treatment on human MSCs evaluated through in-silico pathway analysis in relation to galectin-3.

Soil salinization severely constrains crop productivity worldwide. In cucumber (Cucumis sativus L.), salt stress induces oxidative damage and impairs photosynthesis. Melatonin (MT) improves abiotic stress tolerance across diverse crops; yet its salt tolerance mechanism in cucumber remains unclear.

Septic cardiomyopathy (SCM) is a prevalent and severe complication associated with sepsis. This study explores the effects of sevoflurane and pentobarbital on lipopolysaccharide (LPS) -induced SCM and elucidates underlying mechanisms. The SCM model was established using an intraperitoneal injection of 10 mg/kg LPS. Pentobarbital and sevoflurane were administered thirty minutes post-model establishment. Following the echocardiographic assessment, mice were euthanized 24 h after the modeling, and cardiac samples were collected. Gene sequencing and western blot were utilized to identify potential hub genes and signaling pathways. Sevoflurane markedly reduced LPS-induced myocardial injury and cardiac dysfunction compared to the pentobarbital intervention. Transcriptome sequencing revealed that numerous genes exhibited differential expression following intervention with sevoflurane and pentobarbital, with predominant enrichment in the signaling pathways, such as the extracellular region and matrix, tumor necrosis factor (TNF), and p53 signaling. Sevoflurane significantly induced TATA-box binding protein-associated factor, RNA polymerase I subunit D (TAF1D) expression and attenuated cardiomyocyte death, oxidative stress, and the secretion of IL-6 and TNF-α compared to the pentobarbital group (p < 0.05). Furthermore, oe-TAF1D significantly exacerbated cardiomyocyte death, oxidative stress, and inflammatory responses, which were alleviated with si-TAF1D (p < 0.05). Sevoflurane mitigates sepsis-induced cell death, oxidative stress, and inflammatory responses by up-regulating TAF1D, consequently diminishing cardiac injury and preserving cardiac function.

Glioblastoma, IDH wild-type (WHO grade 4) (GBM), is the most common primary brain tumor in adults with a 21-month median overall survival, despite surgical-resection and radio-chemotherapy. Bevacizumab, a monoclonal antibody towards vascular endothelial growth factor-A, is used to treat recurrent-GBM. To find predictors of poor-response, patient-derived xenograft (PDX)-tumors were treated with bevacizumab or vehicle and subsequently grouped based on survival-response; RNAseq expression was then compared by responder-status. Bioinformatic-analysis demonstrated differential gene expression in tumors from poor-responders (six-PDXs) as compared to tumors from good-responders (three-PDXs), along with upregulation of angiogenesis and collagen gene-sets in poor-responders. Within these gene-sets, multiple genes known to be regulated by the early growth response-1 (EGR1) transcription factor, which was also upregulated, were identified and CHRNA7 (α7-nicotinic-acetylcholine receptor, α7-nAChR) was selected for validation. In terms of protein/functional studies, in the bevacizumab-treated poor-responders, nuclear-EGR1 was elevated, Ki67-labeling was increased in EGR1high tumor, and there was increased angiogenesis. Expression of α7-nAChR and nuclear EGR1 was directly correlated, suggesting CHRNA7 is an EGR1 downstream target. Data-mining (GLASS-database) showed that recurrent GBM in females with an elevated EGR1 and methylated MGMT promoter had a shorter survival. In summary, GBM with increased EGR1 expression, Ki67-labeling in EGR1high tumor and angiogenesis demonstrated a poor-response to bevacizumab, suggesting EGR1 could be useful in predicting response.

Fenofibrate, a peroxisome proliferator-activated receptor α (PPARα) agonist, is widely prescribed to treat hyperlipidemia and has therapeutic potential in liver and kidney diseases. However, fenofibrate is also associated with adverse effects, including elevated creatinine and liver and kidney toxicity, although the underlying mechanisms remain unclear. In addition, how fenofibrate regulates lipid metabolism differently in the liver and kidney is not well understood. Therefore, in this study, we investigated the dose-dependent effects of fenofibrate on liver and kidney metabolism in rats, with a focus on PPARα activation and potential mechanisms contributing to organ-specific toxicity. We used high-throughput transcriptomic data from 5-day rat in vivo studies, where rats were exposed to fenofibrate, and performed pathway enrichment, injury module, and detailed individual gene comparison analyses to investigate how liver and kidney metabolism were differentially altered between the two organs. Fenofibrate exposure significantly increased liver but not kidney weights and caused larger perturbations in the liver compared to the kidney transcriptome, with the majority of the changes related to PPARα regulation. Interestingly, our study revealed that the PPARα and RXRα genes are differentially regulated between the liver and kidney. In addition, we identified several differences between them in cellular and mitochondrial fatty acid transport, lipoprotein metabolism, fatty acid oxidation, branched-chain amino acid degradation, and glucose metabolism pathways. Furthermore, we identified transcriptomic inflection points at which the changes in the PPARα-mediated regulation of lipid metabolism switched from beneficial to deleterious as the fenofibrate concentration increased leading to liver injury, providing potential mechanisms of toxicity.

Chronic benzene exposure causes hematotoxicity, yet the underlying molecular mechanisms remain incompletely understood. N6-methyladenosine (m6A) RNA modification has emerged as a critical regulator in various diseases, but its role in benzene-induced hematotoxicity is unclear. In this study, m6A mRNA microarray analysis revealed widespread alterations in m6A methylation following benzene exposure. Gene Ontology (GO) enrichment identified ubiquitin conjugating enzyme E2 G2 (UBE2G2) as a key m6A-methylated transcript associated with protein degradation pathways. Bioinformatics prediction and experimental validation (RIP-qPCR and MeRIP-qPCR) confirmed that the m6A reader YTHDF3 directly binds to UBE2G2 mRNA in an m6A-dependent manner. Benzene exposure downregulated YTHDF3 expression, leading to decreased UBE2G2 mRNA stability and expression. Overexpression of YTHDF3 reversed this suppression, confirming the regulatory relationship. Functionally, reduced UBE2G2 impaired ubiquitination of acyl-CoA synthetase long-chain family member 4 (ACSL4), resulting in its stabilization and upregulation, which promoted lipid peroxidation and ferroptosis. Notably, melatonin treatment ameliorated benzene-induced hematotoxicity by restoring the YTHDF3/UBE2G2/ACSL4 axis. In summary, this study identifies a novel epigenetic pathway-YTHDF3/m6A/UBE2G2/ACSL4-that mediates benzene-induced ferroptosis, and demonstrates the protective effect of melatonin through modulation of this axis.

Recent studies show that glioblastoma (GBM) is more sensitive to temozolomide (TMZ) in the morning. In cells, inhibiting O6-Methylguanine-DNA-Methyltransferase (MGMT) abolished time-dependent TMZ efficacy, suggesting that circadian regulation of this DNA repair enzyme underlies daily TMZ sensitivity. Here, we tested the hypotheses that MGMT promoter methylation and protein abundance vary with time-of-day in GBM, resulting in daily rhythms in TMZ efficacy.

Global industrialization has made soil heavy metal pollution a critical environmental issue, threatening human health. In many agricultural fields, heavy metal contamination coexists with herbicides, worsening crop safety risks. This study investigated napropamide (NAP) enantiomers' differential effects on plant growth, nutrient, secondary metabolites, and cadmium (Cd). Compared to the control, R-NAP significantly promoted plant growth and increased Cd accumulation, while S-NAP inhibited growth. Moreover, NAP treatments altered the secondary metabolites, particularly flavonoids and quinones. These metabolites might be regulated by macro- and micronutrients, thereby influencing plant Cd content. Furthermore, flavonoid- and quinone-related genes (e.g., OMT1 and 4CL3) were significantly upregulated under R-NAP treatments. Core metabolites strongly associated with these genes and Cd content, confirming their role in Cd accumulation. This work elucidates how NAP enantiomers regulate nutrient-mediated secondary metabolism against Cd stress, offering insights for controlling Cd accumulation in slightly to moderately contaminated soils under herbicide use.

The efficacy of the SGLT2 inhibitor empagliflozin (EMPA) in mitigating hepatic steatosis in patients with type 2 diabetes mellitus and metabolic dysfunction-associated steatotic liver disease (MASLD) has been previously demonstrated. However, the underlying mechanisms remain unclear. In this study, we investigated the role of EMPA in alleviating hepatic steatosis through the modulation of O-GlcNAcylation. High-glucose (HG)-induced alpha mouse liver 12 (AML12) cells, mouse primary hepatocytes (MPHs), and murine MASLD models (high-fat diet-fed and ob/ob mice) were used to examine the effects of EMPA. Protein O-GlcNAcylation, lipid accumulation, and AMP-activated protein kinase α (AMPKα) regulation were evaluated using Western blotting, immunostaining, and siRNA knockdown. Our findings showed that protein O-GlcNAcylation levels were elevated in both in vitro and in vivo models. EMPA treatment reduced O-GlcNAcylation and ameliorated lipid accumulation in HG-induced AML12 cells, MPHs, and MASLD models. Knockdown of O-GlcNAc transferase (OGT) decreased O-GlcNAcylation levels and lipid accumulation in HG-induced AML12 cells. Additionally, OGT knockdown altered both O-GlcNAcylated and phosphorylated AMPKα levels. In these models, EMPA administration decreased O-GlcNAcylated AMPKα while increasing phosphorylated AMPKα. This study further identified serine 344, threonine 447, and serine 501 as critical O-GlcNAcylation sites on AMPKα2. Mutation of these residues to alanine in AMPKα2 attenuated lipid accumulation in AML12 cells, with no additional improvement observed following EMPA treatment. In summary, EMPA effectively improves hepatic steatosis by modulating the O-GlcNAcylation states of AMPKα. Identification of specific O-GlcNAcylation sites on AMPKα2 highlights their importance in the therapeutic mechanism of EMPA in improving hepatic steatosis.

Chronic consumption of a Western-style diet, high in saturated fats and cholesterol, disrupts energy metabolism, leading to obesity and insulin resistance. In contrast, isoflavone (Isof)-rich soy has been shown to benefit connective tissue metabolism. However, the potential of soy Isof dietary supplementation to counteract the effects of a high-fat diet (HFD) on bone development, morphology, and remodeling remains poorly understood. In this study, 24-day-old male Sprague-Dawley rats were fed a HFD containing 45% fat and 0.25% cholesterol for 8 weeks, leading to significant increases in body weight, long bone length, bone marrow adiposity, and insulin resistance when compared to control diet (AIN-93G) rats. Using micro-CT and three-point bending tests, we found that supplementation of HFD with dietary soy Isof (NovasoyR 400: 352 mg Isof/g; Genistein, Daidzein, Glycitein ratio of 1.3:1:0.15 identical to that found in soybeans) prevented HFD-induced reductions in bone mass, including bone volume, trabecular number, density, and strength. These bone-preserving effects were associated with decreased non-esterified free fatty acid (NEFA) levels and increased alkaline phosphatase activity in serum and bone marrow plasma. Isof supplementation also ameliorated HFD-induced increases in the expression of Ezh2 and H3k27me3 levels (gene silencing epigenetic mark catalyzed by Ezh2) in bone and blocked the elevated expression of osteoclastic marker NFATc1 in bone marrow cells. A significant inverse correlation between bone mineral density (BMD) and DNA methylation marker 5-methylcytosine (%5-mC), as well as a positive correlation between serum NEFA levels and Ezh2 expression, were observed. Furthermore, Isof supplementation inhibited HFD-induced increases in bone DNA methylation. These results suggest that: (1) HFD disrupts bone and adipose tissue development and remodeling in post-weanling juvenile rat models; (2) Isofs may protect against high-fat diet-induced bone impairments. We hypothesize that the molecular mechanisms underlying Isof's bone protective effects may involve modulation of histone and DNA methylation through regulation of Ezh2 levels.

Kaposiform haemangioendothelioma (KHE) research faces challenges due to the lack of established cell lines and suitable animal models. Our study aimed to establish KHE cell lines, spheroids and refine murine models to mimic disease characteristics, advancing our understanding of KHE pathogenesis and exploring novel therapies. Primary KHE cells were sorted using CD31 antibodies and cultured into spheroids. These cells were then injected into mice, and the resulting tumours were analysed using immunohistochemistry. Preliminary exploration of the potential mechanisms of sirolimus action on KHE was conducted through transcriptome sequencing. CD31+ KHE cells were isolated and characterised from three out of six patients. The CD31+ KHE cells demonstrated positive expression of essential markers such as CD31, Ki67 and LYVE1, consistent with the profiles observed in KHE tumours. Additionally, subcutaneous tumours displayed similar positive expression of key markers, reminiscent of KHE tumours. Transcriptome sequencing revealed downregulation of ATG9B after sirolimus treatment in CD31+ KHE cells. CD31+ KHE cells can replicate human KHE in murine models, offering a valuable tool for studying pathogenesis. Our findings also suggest a potential mechanism of sirolimus action in treating KHE, warranting further investigation into novel therapeutic strategies.

Both high-cholesterol diet (HCD) and fine particulate matter (PM2.5) exposure are associated with increased stroke risk; however, their combined effects-particularly in females-remain poorly understood. This study aimed to investigate the synergistic effects of HCD and PM2.5 exposure on brain injury, behavior, and underlying molecular mechanisms in female mice. Female mice were exposed to HCD, PM2.5 inhalation, or both for 3 months (3M) or 6 months (6M), with control groups for comparison. We assessed brain-to-body weight ratio, behavioral performance, histopathological changes, inflammatory markers (IL-6, TNF-α), and expression of stroke- and blood-brain barrier (BBB)-related proteins. Additionally, we analyzed the epigenetic regulation of hypoxia-inducible factor-1α (HIF-1α) via chromatin immunoprecipitation (ChIP) assays. Co-exposure to HCD and PM2.5 led to greater alterations in the brain-to-body weight ratio, exacerbated neuropathology, behavioral deficits, and increased neuroinflammation compared to exposure to either factor alone. Moreover, HCD+PM2.5 significantly altered the expression of HIF-1α and genes involved in its downstream signaling pathway in the brain. ChIP assays revealed that HIF-1α was directly regulated by histone deacetylase 4 (HDAC4) and histone H3 lysine 9 acetylation (H3K9ac), indicating epigenetic modulation in response to dual exposure. HCD and PM2.5 synergistically promote brain injury in female mice, potentially through HDAC4/H3K9ac-mediated epigenetic activation of the HIF-1α pathway. These findings highlight a novel molecular mechanism that may contribute to increased stroke risk in females exposed to both environmental and metabolic stressors.