FoxO Signaling Research Articles

Comparative Analyses of Dynamic Transcriptome Profile of Heart Highlight the Key Response Genes for Heat Stress in Zhikong Scallop Chlamys farreri.

Heat stress resulting from global climate change has been demonstrated to adversely affect growth, development, and reproduction of marine organisms. The Zhikong scallop (Chlamys farreri), an important economical mollusk in China, faces increasing risks of summer mortality due to the prolonged heat waves. The heart, responsible for transporting gas and nutrients, is vital in maintaining homeostasis and physiological status in response to environmental changes. In this study, the effect of heat stress on the cardiac function of C. farreri was investigated during the continuous 30-day heat stress at 27 °C. The results showed the heart rate of scallops increased due to stress in the initial phase of high temperature exposure, peaking at 12 h, and then gradually recovered, indicating an acclimatization at the end of the experiment. In addition, the levels of catalase (CAT), superoxide dismutase (SOD), and total antioxidant capacity (T-AOC) exhibited an initial increase followed by recovery in response to heat stress. Furthermore, transcriptome analysis of the heart identified 3541 differentially expressed genes (DEGs) in response to heat stress. Subsequent GO and KEGG enrichment analysis showed that these genes were primarily related to signal transduction and oxidative stress, such as the phosphatidylinositol signaling system, regulation of actin cytoskeleton, MAPK signaling pathway, FoxO signaling pathway, etc. In addition, two modules were identified as significant responsive modules according to the weighted gene co-expression network analysis (WGCNA). The upregulation of key enzymes within the base excision repair and gap junction pathways indicated that the heart of C. farreri under heat stress enhanced DNA repair and maintained cellular integrity. In addition, the variable expression of essential signaling molecules and cytoskeletal regulators suggested that the heart of C. farreri modulated cardiomyocyte contraction, intracellular signaling, and heart rate through complex regulation of phosphorylation and calcium dynamics in response to heat stress. Collectively, this study enhances our understanding of cardiac function and provides novel evidence for unraveling the mechanism underlying the thermal response in mollusks.

Employing Multi-Omics Analyses to Understand Changes during Kidney Development in Perinatal Interleukin-6 Animal Model.

Chronic kidney disease (CKD) is a leading cause of morbidity and mortality globally. Maternal obesity during pregnancy is linked to systemic inflammation and elevated levels of the pro-inflammatory cytokine interleukin-6 (IL-6). In our previous work, we demonstrated that increased maternal IL-6 during gestation impacts intrauterine development in mice. We hypothesized that IL-6-induced inflammation alters gene expression in the developing fetus. To test this, pregnant mice were administered IL-6 or saline during mid-gestation. Newborn mouse kidneys were analyzed using mRNA-seq, miRNA-seq and whole-genome bisulfite-seq (WGBS). A multi-omics approach was employed to quantify mRNA gene expression, miRNA expression and DNA methylation, using advanced bioinformatics and data integration techniques. Our analysis identified 19 key genes present in multiple omics datasets, regulated by epigenetics and miRNAs. We constructed a regulatory network for these genes, revealing disruptions in pathways such as Mannose type O-glycan biosynthesis, the cell cycle, apoptosis and FoxO signaling. Notably, the Atp7b gene was regulated by DNA methylation and miR-223 targeting, whereas the Man2a1 gene was controlled by DNA methylation affecting energy metabolism. These findings suggest that these genes may play a role in fetal programming, potentially leading to CKD later in life due to gestational inflammation.

Hederagenin regulates the migration and invasion of hepatocellular carcinoma cells through FOXO signaling pathway.

This study aimed to elucidate the effects of Hederagenin (HG) on hepatocellular carcinoma (HCC) and explore its potential molecular mechanisms. Virtual screening was employed to identify potential targets within core pathways of liver cancer and to analyze the possible mechanisms of HG. CCK-8 assays were used to assess the viability of HCC cells, while Hoechst 33342/PI staining was utilized to evaluate apoptosis. The migration and invasion abilities of HCC cells were examined using Transwell and scratch assays, and single-cell cloning ability was assessed via colony formation assays. Subsequent qRT-PCR was conducted to determine the mRNA expression levels of FOXO1 and FOXO6 following HG treatment. Western blot (WB) analysis was employed to measure the protein expression levels of IGF1R, FOXO1, FOXO6, MMP2, MMP9, and VEGFA, as well as the phosphorylation status of FOXO1 Ser249. Virtual screening indicated that HG might exert antitumor effects through the FOXO signaling pathway. Experimental results demonstrated that HG induces apoptosis in a dose-dependent manner and inhibits the proliferation, migration, invasion, and single-cell cloning ability of HCC cells. After HG treatment, FOXO1 expression was upregulated, while the expression levels of IGF1R, phosphorylated FOXO1 Ser249, MMP2, MMP9, and VEGFA were downregulated. In summary, our study is the first to demonstrate that HG regulates the phosphorylation of FOXO1, affecting the proliferation, migration, and invasion of HCC cells. The findings suggest that HG can inhibit the migration of HCC cells in vitro. The data indicate that HG-mediated targeting of the FOXO1/FOXO6 pathway holds promise as a novel therapeutic approach.

Dietary supplementation with sodium propionate and tributyrin alleviated hepatic lipid deposition and improved the antioxidant capacity and hypoxic stress resistance of spotted seabass (Lateolabrax maculatus)

We investigated the effects of dietary supplementation with sodium propionate (SP) and tributyrin (TB) on hepatic lipid deposition and antioxidant capacity of spotted seabass (Lateolabrax maculatus) via an 8-week feeding experiment and a hypoxia stress experiment. The fish were fed five experimental diets: a control diet (CON), a diet supplemented with 2 g/kg SP (SP-0.2%), 4 g/kg SP (SP-0.4%), 2 g/kg TB (TB-0.2%), or 4 g/kg TB (TB-0.4%). No significant difference in growth performance was presented among the groups (P > 0.05). The SP-0.4% and TB-0.2% groups presented significantly lower hepatosomatic and viscerasomatic indexes compared with the CON group. Then, the SP-0.4% and TB-0.2% groups presented stronger resistance to hypoxic stress than the other groups and were analyzed further. The hepatic histology and triglyceride levels revealed that SP-0.4% and TB-0.2% reduced hepatic lipid deposition. Similarly, the downregulation of malondialdehyde and the upregulation of total antioxidant capacity, superoxide dismutase, and catalase activities and the related gene expression levels revealed that SP-0.4% and TB-0.2% improved the antioxidant capacity. Additionally, the RNA sequencing demonstrated that SP-0.4% and TB-0.2% regulated gene expression to a similar extent. Among the 117 differentially expressed genes, 67 genes were enriched in the same pattern, and involved the FoxO signaling, PI3K-Akt signaling, and insulin-related pathways. In conclusion, supplementing SP-0.4% and TB-0.2% as feed additives effectively improved hepatic lipid metabolism, antioxidant capacity, and hypoxic stress resistance of spotted seabass.

The effects of microplastics exposure on quail's hypothalamus: Neurotransmission disturbance, cytokine imbalance and ROS/TGF-β/Akt/FoxO3a signaling disruption

Microplastics (MPs) have become a major focus of environmental toxicology, raising concerns about their potential adverse effects on animal organs and body systems. As these tiny particles infiltrate ecosystems, they may pose risks to the health of organisms across diverse species. In this study, we attempted to examine the neurotoxic effects of MPs exposure on avian hypothalamus by using an animal model-Japanese quail (Coturnix japonica). The quails of 7-day-old were exposed to 0.02 mg/kg, 0.4 mg/kg and 8 mg/kg polystyrene microplastic (PS-MPs) of environmental relevance for 35 days. The results showed PS-MPs exposure did damages to hypothalamic structure characterized by neuron malformation, irregular arrangement and cellular vacuolation after 5-week exposure. PS-MPs exposure also induced Nissl body reduction and dissolution in the hypothalamus. Moreover, the decrease of acetylcholinesterase (AchE) activity and increasing acetylcholine (Ach) indicated that PS-MPs exposure caused hypothalamic neurotransmission disturbance. PS-MPs exposure also led to neuroinflammation by disrupting the balance between proinflammatory and anti-inflammatory cytokines. Moreover, increasing reactive oxygen species (ROS) and malondialdehyde (MDA) generation with reducing antioxidants indicated PS-MPs led to hypothalamic oxidative stress. Additionally, RNA-Seq analysis found that both transforming growth factor-β (TGF-β) signaling and forkhead box O (FoxO) signaling were disturbed in the hypothalamus by PS-MPs exposure. Especially, the increasing ROS led to TGF-β activation and then induced hypothalamic inflammation by nuclear factor κB (NF-κB) activation. The present study concluded that oxidative stress might be an important mechanistic signaling involved in MPs neurotoxicology.

Network Pharmacology Integrated with Pharmacological Evaluation for Investigating the Mechanism of Resveratrol in Perimenopausal Depression

Perimenopause constitutes a pivotal transitional phase characterized by hormonal variability and heightens vulnerability to depressive episodes. This study seeks to elucidate the mechanism of resveratrol (RES) in perimenopausal depression through integrated network pharmacology, molecular docking analysis, and experimental validation. Screening yielded 83 RES-related disease targets, with IL10, CCL2, and SERPINE1 identified as core genes overexpressed in perimenopausal depression. GO analysis and KEGG pathway enrichment analysis predicted that the target genes could regulate the PI3K-Akt, FoxO, HIF-1, and IL-17 signaling pathways. Molecular docking indicated SERPINE1 as a promising RES target. Consistently, in vitro experiments showed that RES significantly attenuated the inflammatory response and apoptosis of lipopolysaccharide-stimulated CTX-TNA2 cells. RES also reduced the expression of NLRP3, caspase-1, SERPINE1 proteins and acetylation, while increasing the expression of BDNF, TrkB, SIRT1, and decreasing MAO-A proteins. In vivo experiments demonstrated that RES also significantly improved the depression behaviors, increased the levels of 5-HT1A and SIRT1, and decreased levels of MAO-A of depression rats. This study unveils RES's potential targets and mechanism in perimenopausal depression, laying groundwork for future research.

The Effects of Heat Therapy During Immobilization and Rehabilitation on Muscle Atrophy and Strength Loss at Return to Sports in Healthy Humans.

Animal research suggests that repeated heat exposures may stimulate skeletal muscle protein synthesis and downregulate protein degradation. Repeated heat exposures during ankle immobilization and rehabilitation would preserve human muscle strength and mass. Controlled laboratory study. A total of 20 male participants (age, 33.6 ± 2.8 years; weight, 83.8 ± 9.2 kg; height, 182 ± 6 cm) underwent 4 weeks of supervised training, 2 weeks of single-lower leg immobilization, and 2 weeks of supervised rehabilitation before return to sports (RTS). Participants were split into 2 groups: (1) whole-body heat therapy (HEAT) and (2) sham treatment (SHAM) throughout the immobilization and rehabilitation periods. Measures of muscle strength (isometric and isokinetic), volume (magnetic resonance imaging and ultrasound), and muscle biopsies were obtained preimmobilization, postimmobilization, and at RTS. Maximal isometric strength of the plantarflexors was lower at RTS compared with preimmobilization in SHAM (P = .027) but not HEAT (P = .301). Isokinetic strength during a fatigue test was higher at RTS compared with preimmobilization in HEAT (P = .039) but not SHAM (P = .245). Pennation angle and muscle thickness were lower at postimmobilization compared with preimmobilization only in SHAM (P≤ .027). Muscle cross-sectional area decreased in soleus and both gastrocnemius medialis and lateralis (all P≤ .035) in SHAM, but only in gastrocnemius medialis in HEAT. There was a large (d = 0.91) but not significant (P = .054) decrease in the ratio of phosphorylated/total nuclear factor-kappa B (NFκB) from preimmobilization to postimmobilization in HEAT only. There was an increase in phosphorylated fork head box O proteins (FoxO) only in HEAT (P = .034), suggesting a decrease in FoxO activity. Caspase 3 expression increased from preimmobilization to postimmobilization in SHAM only (P = .004). These results indicate that using heat therapy throughout immobilization and rehabilitation reduces skeletal muscle atrophy and maintains plantarflexor strength in healthy humans. Moreover, heat therapy may lead to the inactivation of the FoxO and NFκB signaling pathways involved in atrophy. Repeated heat exposures should be considered a novel therapeutic intervention to counteract muscle atrophy during immobilization.

Profiling genetic variants in cardiovascular disease genes among a Heterogeneous cohort of Mendelian conditions patients and electronic health records.

This study addresses the rising cardiovascular disease (CVD) rates in the United Arab Emirates (UAE) by investigating the occurrence and impact of genetic variants in CVD-related genes. We collected all genes linked to heritable CVD from public and diagnostic databases and mapped them to their corresponding biological processes and molecular pathways. We then evaluated the types and burden of genetic variants within these genes in 343 individuals from the Emirati Mendelian Study Cohort and 3,007 national electronic health records. We identified a total of 735 genes associated with heritable CVD, covering a range of cardiovascular conditions. Enrichment analysis revealed key biological processes and pathways, including Apelin, FoxO, and Ras signaling, that are implicated across all forms of heritable CVD. Analysis of a UAE cohort of 3,350 individuals showed a predominance of rare and unique CVD variants specific to the population. The study found a significant burden of pathogenic variants in families with CVD within the Emirati Mendelian cohort and re-assessed the pathogenicity of 693 variants from national health records, leading to the discovery of new CVD-causing variants. This study underscores the importance of continuously updating our understanding of genes and pathways related to CVD. It also highlights the significant underrepresentation of the UAE population in public databases and clinical literature on CVD genetics, offering valuable insights that can inform future research and intervention strategies.

MiR-132-3p downregulates FOXO1 in CD4+ T cells and is associated with disease manifestations in patients with lupus.

This study aimed to evaluate the expression status of miR-132-3p in CD4+ T cells in patients with systemic lupus erythematosus (SLE) and explore its potential role in SLE development. The study included 60 patients with SLE and 30 healthy controls. miR-132-3p expression in CD4+ T cells was detected by real-time quantitative reverse transcription polymerase chain. Bioinformatics analyses were employed to predict target genes and explore the potential role of miR-132-3p. The associations between miR-132-3p levels and SLE Disease Activity Index (SLEDAI) score, as well as laboratory characteristics, were analyzed. miR-132-3p levels in CD4+ T cells were significantly higher in patients with SLE compared with healthy controls. Bioinformatics analysis identified FOXO1 as a potential target gene of miR-132-3p, with a particular emphasis on the FOXO signaling pathway. miR-132-3p up-regulation in CD4+ T cells was associated with high SLEDAI score, high anti-double-stranded DNA levels, low C3 and C4 levels, positive anti-ribosomal P, and high 24-hour urinary protein levels in patients with SLE. miR-132-3p may contribute to CD4+ T cell dysregulation during SLE by targeting FOXO1 and could potentially be used to assess disease severity.

Dietary Lithospermum erythrorhizon ethanol extract alleviates soybean meal-induced enteritis by improving immune tolerance profile of pearl gentian grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂)

The aquaculture industry has a shortage of objected protection against soybean meal-induced enteritis (SBMIE) in carnivorous fish caused by soybean meal feed. Our initial study discovered that Lithospermum erythrorhizon (LE) ethanol extract has potential application value in improving SBMIE. A feeding trial (for eight weeks) was conducted to investigate LE ethanol extract on pearl gentian grouper SBMIE of protection to clarify the influence of LE ethanol extract on the immune tolerance profile. Three hundred and sixty pearl gentian groupers were administered one of three distinct dietary regimes: 1) 100 % fish meal (FM); 2) soybean meal substitution of 50 % fish meal protein (SBM); 3) SBM diet +0.2 % LE ethanol extract (SBMLE). Each treatment included three 1000 L cisterns—each of cisterns with 30 fish. The preliminary weight of the fish varied between 72.01 g and 72.50 g. Growth performance results showed that WGR and SGR were significantly decreased in the SBM group (P < 0.05), while there was no significant difference between the FM and SBMLE groups. There was no significant difference in survival among the three groups. The results showed that SBM-fed fish exhibited enteritis manifested by mucosal fold shortening, lamina propria widening, decreased serum immune markers (IgM, C3, and C4), and up-regulated expression of pro-inflammatory cytokines (il17 and il12) and immune-related gene (tlr3, and tlr9). The addition of 0.2 % LE ethanol extract to the SBM diet, reversed the above symptoms, and anti-inflammatory cytokine (tgf-β1), gene expression increased significantly (P < 0.05). Intestinal transcriptome analysis exhibited that the DEGs between the FM group and the SBM group were mainly enriched in FoxO signaling pathway, while the DEGs between the SBM group and the SBMLE group were enriched in MAPK signaling pathway and FoxO signaling pathway. The RT-qPCR results also revealed changes in MAPK/FoxO signaling pathway-related genes, including Dusp1, jund, Irs2b, fbxo32, and ccng2. Overall, Lithospermum erythrorhizon ethanol extract may alleviate SBMIE by regulating MAPK/FoxO signaling pathway, which would be beneficial for enhancing the immune tolerance and utilization efficiency of pearl gentian groupers to dietary soybean meal.

Transcriptomic Study of Different Stages of Development in the Testis of Sheep.

Numerous genes govern male reproduction, modulating testicular development and spermatogenesis. Our study leveraged RNA-Seq to explore candidate genes and pivotal pathways influencing fecundity in an F1 hybrid of Southdown × Hu sheep testes across four developmental milestones: M0 (0 months old, newborn), M3 (3 months old, sexually immature), M6 (6 months old, sexually mature), and Y1 (1 years old, adult). Histological examination using hematoxylins and eosin staining revealed that the cross-sectional area of the spermatid tubules and the number of supportive cells increased in the other groups, as compared to the M0 group. The cross-sectional area of the vasculature and the number of supporting cells were found to be significantly increased in all other groups in comparison to the M0 group. We conducted GO and KEGG analyses of the differentially expressed genes (DEGs) in the three comparison groups and identified key pathways, including cAMP, MAPK, ECM-receptor interactions, PI3K-Akt, and FOXO signaling, which are closely related to testicular development and spermatogenesis. Notably, alternative splicing (AS) events were markedly elevated in M6 and Y1 stages. Key genes like GATA4, GATA6, SMAD4, SOX9, YAP1, ITGB1 and MAPK1 emerged as significantly enriched in these pathways, potentially orchestrating the transition from immature to mature testes in sheep. These findings offer valuable insights into male reproductive potential and can inform strategies for optimizing animal breeding.

Identification of crucial genes and possible molecular pathways associated with active vitamin D intervention in diabetic kidney disease

BackgroundA significant cause of advanced renal failure is diabetic nephropathy (DKD), with few treatment options available. Calcitriol shows potential in addressing fibrosis related to DKD, though its molecular mechanisms remain poorly understood. This research seeks to pinpoint the crucial genes and pathways influenced by calcitriol within the scope of DKD-related fibrosis. MethodsSingle-cell gene expression profiling of calcitriol treated DKD rat kidney tissue and screening of fibrosis-associated cell subsets. Mendelian randomization and enrichment analyses (CIBERSORT, GSVA, GSEA, Motif Enrichment) were used to explore gene-immune cell interactions and signaling pathways. Key findings were validated using independent datasets and protein expression data from the Human Protein Atlas. ResultsCalcitriol treatment reduced proliferative cell populations and highlighted the FoxO signaling pathway's role in DKD. SUMO3 and CD74 were identified as key markers linked to immune infiltration and renal function. These genes were significantly associated with creatinine levels and eGFR, indicating their potential role in DKD progression. ConclusionOur results suggest that calcitriol modulates DKD fibrosis through the FoxO pathway, with SUMO3 and CD74 serving as potential biomarkers for kidney protection. These results provide fresh insights into strategies for treating DKD.

Effect and mechanism of the PTMAP5–hsa-miR-22-3p–KIF2C regulatory axis on the occurrence and development of hepatocellular carcinoma

Hepatocellular carcinoma (HCC), a primary liver cancer, is known for its high malignancy potential. Despite extensive research, the etiology of HCC remains elusive, with numerous molecular mechanisms yet to be deciphered. This study delves into the influence and mechanism of the PTMAP5&amp;ndash;hsa-miR-22-3p&amp;ndash;KIF2C regulatory axis in HCC pathogenesis. A comparative analysis of gene expression profiles between the GSE87630 and GSE45267 datasets unveiled a cohort of 346 differentially expressed genes. Protein&amp;ndash;protein interaction networks were established using the STRING database, and key genes were identified through receiver operating characteristic curve analysis and LASSO regression analysis. The expression and prognostic value of these key genes were further evaluated using GEPIA and Kaplan&amp;ndash;Meier analysis. Upstream miRNAs were predicted using the MiRTarBase and StarBase databases, facilitating the construction of the PTMAP5&amp;ndash;hsa-miR-22-3p&amp;ndash;KIF2C regulatory axis. Co-expression analysis of KIF2C was performed through UALCAN and StarBase, and the regulatory axis was found to play a pivotal role in HCC development through the cell cycle, oocyte meiosis, and FOXO signaling, as revealed by DAVID enrichment analysis. Furthermore, the expression and prognostic significance of KIF2C in various cancer types were assessed using TIMER and GEPIA, while TISIDB analysis revealed correlations between KIF2C expression and relevant major histocompatibility complex molecules, immunomodulators, chemokines, receptors, and immune variants in HCC. These findings also extended to HCC molecular subtypes. In conclusion, we constructed a novel PTMAP5&amp;ndash;hsa-miR-22-3p&amp;ndash;KIF2C regulatory subnetwork, which could provide valuable therapeutic targets and diagnostic markers for HCC.

MiR-98 and miR-629 can be used as a potential biomarker on relapsing-remitting multiple sclerosis patients

IntroductionMultiple Sclerosis (MS) is an autoimmune disease that affects the central nervous system (CNS), characterized by chronic inflammation, demyelination, and neurodegeneration. In recent years, among non-coding RNAs miRNA have emerged as key regulators of different biological processes and it has been suggested that they play an important role in the mechanisms underlying MS pathogenesis. Through in silico methods, miR-629-5p and miR-98-5p were identified as significant factors in MS pathology. The aim of this study was to examine the levels of expression of miR-629-5p and miR-98-5p in blood samples obtained from patients with relapsing–remitting MS. Methods: Total blood were recruited from RRMS and control group and qPCR analysis was used for evaluating of target miRNAs expression. The mirDIP database was utilized to identify the target genes. Hub genes were identified with the Cytoscape and target pathways were identified using the STRING. Results: Expression analysis revealed a significant upregulation of miR-629-5p and miR-98-5p (FC ≥ 1.5 and p < 0.05) in patients with RRMS. PI3K-Akt, Rap1, MAPK and FoxO signaling pathways were found as target. Discussion: The discovery of these miRNAs suggests that they may have a notable impact on MS patients by intervening on pathways involved in both the immune and nervous systems.

Genome-Wide Association Study of Resistance to Largemouth Bass Ranavirus (LMBV) in Micropterus salmoides.

The disease caused by Largemouth bass ranavirus (LMBV) is one of the most severe viral diseases in largemouth bass (Micropterus salmoides). It is crucial to evaluate the genetic resistance of largemouth bass to LMBV and develop markers for disease-resistance breeding. In this study, 100 individuals (45 resistant and 55 susceptible) were sequenced and evaluated for resistance to LMBV and a total of 2,579,770 variant sites (SNPs-single-nucleotide polymorphisms (SNPs) and insertions-deletions (InDels)) were identified. A total of 2348 SNPs-InDels and 1018 putative candidate genes associated with LMBV resistance were identified by genome-wide association analyses (GWAS). Furthermore, GO and KEGG analyses revealed that the 10 candidate genes (MHC II, p38 MAPK, AMPK, SGK1, FOXO3, FOXO6, S1PR1, IL7R, RBL2, and GADD45) were related to intestinal immune network for IgA production pathway and FoxO signaling pathway. The acquisition of candidate genes related to resistance will help to explore the molecular mechanism of resistance to LMBV in largemouth bass. The potential polymorphic markers identified in this study are important molecular markers for disease resistance breeding in largemouth bass.

Mechanistic and dynamic insight into novel IL7 receptor activators as immunotherapy for the treatment of tuberculosis

The bacterium that causes tuberculosis (TB), Mycobacterium tuberculosis, is still a significant global public health problem due to its contagious nature and the emergence of drug-resistant variants. This study leverages network pharmacology and computational drug discovery to identify genes linked to TB using the GSE54992 and GSE62525 datasets. In active tuberculosis patients, it identifies 162 DEGs that are not present in healthy people. The research includes an investigation of gene ontology (GO) and protein–protein interaction (PPI) networks to provide a comprehensive understanding of these DEGs. Crucially, it identifies ten hub genes: IL7R, CCR7, CXCR5, CCR6, CCR9, S1PR1, BACH2, CXCL5, SPIB, and EBF. The research suggests IL7R as a potential therapeutic target because of its essential role in FoxO signalling pathways and cytokine-cytokine receptor interaction. IL7 and its receptor (IL7R) are vital for T cell functionality and essential in combating TB. The study explores novel small-molecule activators for IL7R (IL7Rα), aiming to boost the immune response against TB. The following four compounds were found using molecular dynamics simulation, virtual screening, computational pharmacophore modelling, and MM/GBSA analytical techniques: ZINC02131028, ZINC02135418, ZINC46007405, and ZINC59273354. These activators may enhance the IL7 signalling pathway, potentially strengthening the immune fight against M. tuberculosis. Among these ZINC02131028 showed the best results to activate IL7 by binding to its active site, stabilizing its active conformation and enhancing its interaction with the IL7 receptor (IL7R). The findings encourage experimental validation of these IL7 activators, marking a significant step towards innovative TB treatment strategies.

Elucidating ferroptosis mechanisms in heart failure through transcriptomics, single-cell sequencing, and experimental validation

BackgroundThe mechanisms underlying ferroptosis in heart failure (HF) remain incompletely understood. MethodsThis study analyzed the heart failure dataset from the Gene Expression Omnibus to identify differentially expressed ferroptosis-related genes (DFRGs). Key DFRGs were selected using LASSO regression and SVM-RFE machine learning techniques. Their diagnostic accuracy was evaluated via ROC curve analysis. Single-cell sequencing data, heart failure cell, and mouse models were utilized to validate these key DFRGs. Additionally, potential non-coding RNAs targeting these genes were predicted, and analyses for gene set enrichment, immune cell infiltration, and drug targeting were conducted. ResultsA total of 127 DFRGs were identified, with 83 downregulated and 44 upregulated compared to controls. Seven key DFRGs (PTGS2, BECN1, SLC39A14, QSOX1, MLST8, TMSB4X, KDM4A) were identified, showing high diagnostic accuracy (AUC 0.988) in the GSE5406 dataset. GO and KEGG analyses linked these genes to ferroptosis, FoxO signaling, and autophagy pathways. A ceRNA network identified 217 miRNAs and 243 lncRNAs potentially targeting these genes, and 64 drugs were predicted as potential targets. Single-cell sequencing and in vitro experiments revealed differential expression of SLC39A14 and QSOX1, which was further confirmed in vivo. ConclusionThis study provides novel insights into the role of ferroptosis in heart failure by identifying and validating DFRGs that exhibit differential expression across various cell types. The differential expression patterns of these genes, particularly SLC39A14 and QSOX1, indicate their potential involvement in the pathophysiological mechanisms contributing to HF. These findings offer new insights for the development of targeted therapies for HF.

Abstract B006: The role of tumor specific IGFBP-3 in the onset and progression of skeletal muscle wasting in a murine model of pancreatic cancer

Abstract Background: Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of cancer- related death and is highly associated with skeletal muscle wasting (SMW) that leads to treatment intolerance and reduced survival. Research substantiates the role of increased levels of Insulin-like Growth Factor Binding Protein-3 (IGFBP-3) in PDAC-related SMW. Canonically, supraphysiological levels of IGFBP-3 block insulin-like growth factor-1 (IGF-1) binding, thus downregulating the activation of its downstream targets Akt and mTOR. Non-canonical signaling via TGF-βRI results in aberrant FoxO1 signaling, which upregulates the ubiquitin proteasome pathway (UPP) and induces autophagy. Dysregulated UPP and autophagy activation have been shown to promote muscle degradation. Recent findings from our lab implicate canonical and non-canonical signaling of tumor associated IGFBP-3 in increased protein catabolism and decreased anabolism, resulting in SMW in a syngeneic murine model of PDAC. We tested the hypothesis that genetic ablation of IGFBP-3 in PDAC tumor cells attenuates SMW via regulation of Akt and FoxO1, leading to decreases in UPP and autophagy in skeletal muscle.Methods: C57BL/6J female mice (6-8 weeks) were randomized to one of three groups: 1) no tumor control (NTC) (n=10), 2) KCKO-Luc parental cells (PDAC) (n=14), or 3) IGFBP-3-/- KCKO-Luc (n=14). Tumor cells were injected orthotopically. Tumor and lean mass were assessed longitudinally via dual energy x-ray absorptiometry (DEXA). At sacrifice, quadriceps muscles were collected for protein and transcriptional analysis, Tibialis Anterior (TA) muscles for histology and serum for ELISA. TA frozen sections were stained with Oil Red O (ORO) to highlight intramuscular adipogenesis.Results: PDAC mice experience significantly reduced survival (T1/2=56 days) compared to IGFBP-3-/- mice that maintained 100% survival at day 100 (p&amp;lt;0.0001). IGFBP-3-/- mice lost significantly less lean mass from baseline to day 56 compared to PDAC mice (p&amp;lt;0.0001). PDAC mice exhibited 3-fold elevated serum levels of IGFBP-3 compared to NTC and IGFBP-3-/- mice (p&amp;lt;0.01). Transcriptionally, PDAC mice displayed increased expression of igfbp3, tgfbr1, and UPP associated genes fbxo32, and trim63 compared to NTC and IGFBP-3-/- mice. Further, the ratio of pAkt (Ser473), pS6 (Ser235/236), and pFoxO1 (Ser253) to total protein levels were significantly reduced in PDAC mice (p&amp;lt;0.05 vs NTC and IGFBP-3-/-). Additionally, protein levels of ULK1 and the ratio of LC3bII over LC3bI, markers of autophagy, are elevated in PDAC mice compared to NTC and IGFBP-3-/- mice (p&amp;lt;0.05). Lastly, there is a significant increase in ORO positive staining in PDAC mice compared to NTC and IGFBP-3-/- mice (p&amp;lt;0.0001).Conclusion: These findings suggest that PDAC-related SMW is driven by tumor derived IGFBP-3, which acts as an endocrine factor on skeletal muscle cells to inhibit protein synthesis and activate FoxO1 signaling to upregulate the UPP and autophagy. Lastly, we identify skeletal muscle adipogenesis as a biomarker for autophagy upregulation and SMW. Citation Format: Zachary R Sechrist, Calvin L Cole. The role of tumor specific IGFBP-3 in the onset and progression of skeletal muscle wasting in a murine model of pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr B006.

Investigating transcriptomics and metabolomics differences offers insights into the mechanisms of muscular fat deposition in common carp (Cyprinus carpio)

With the global population increasing and lifestyle improving, the demand for high-quality nutritional aquatic foods has been rising. Muscle fat is a crucial nutritional index for evaluating the quality of fish flesh. However, the comprehensive and systematic understanding of the molecular mechanism underlying differences in muscle fat deposition remains insufficient. In this study, we integrated transcriptomics and metabolomics of selected samples with extremely high and low muscle fat in common carp (Cyprinus carpio), the major freshwater aquaculture fish in Asia, to identify critical genes, metabolites and metabolic pathways. We totally identified 204 differentially expressed genes (DEGs) and 1528 differentially accumulated metabolites (DAMs). Glycerolipid, glycerophospholipid and glyoxylate and dicarboxylate metabolisms were enriched through both transcriptomics and metabolomics. These lipid metabolism pathways may be regulated by some critical signal transduction pathways, including Extracellular matrix [ECM]-receptor interaction, mTOR signaling pathway and FoxO signaling pathway. Combined with the validation of gene expression and biochemical indices, a supposed regulatory network was established. To our knowledge, it is the first study to apply a multi-omics approach in fish with naturally different muscle fat to comprehensively elucidate the mechanism. This study could deepen our understanding of the molecular mechanism of muscle fat deposition and be helpful for improving the quality of fish.

MiR-9-5p regulates Sirt1 involved in testicular development and spermatogenesis in mouse

Testicular development and spermatogenesis are critical for male reproduction, with histone (de)acetylation playing a key role in chromatin remodeling within germ cells. Sirt1, a key histone deacetylase, is implicated in chromatin remodeling, but its expression pattern and specific role in testicular development and spermatogenesis need further study. This study comprehensively analyzed Sirt1 expression in adult and juvenile mouse testicular tissues and across various male germ cells, utilizing RT-qPCR, Western blot, immunofluorescence, and cell transfection. GO and KEGG enrichment analyses were performed to elucidate the biological functions and pathways associated with Sirt1 and its related genes. Multiple miRNA databases were utilized to predict miRNAs targeting Sirt1, and their expression levels were validated using RT-qPCR. Lentiviral transfection was used to knockdown candidate miRNAs to assess their functional roles. The results revealed a significant downregulation of Sirt1 expression in adult mouse testicular tissues compared to juvenile tissues, with pronounced variation across diverse male germ cells. Sirt1 was highly expressed in spermatogonia and mature sperm, but comparatively lower in spermatocytes and spermatids. GO and KEGG enrichment analyses highlighted Sirt1's role in key biological processes, including chromatin organization, regulation of cell proliferation, and energy homeostasis, as well as its association with signaling pathways like cellular senescence, the FoxO signaling pathway, and the AMPK signaling pathway. Bioinformatic analysis and subsequent RT-qPCR validation identified miR-9-5p as a miRNA targeting Sirt1. The expression of miR-9-5p was significantly higher in adult mouse testicular tissues compared to juvenile tissues, inversely correlating with Sirt1 levels. Moreover, the knockdown of miR-9-5p led to a notable increase in Sirt1 mRNA and protein expression. In conclusion, Sirt1 is a key player in mouse testicular development and spermatogenesis. The discovery that miR-9-5p negatively regulates Sirt1 suggests a critical regulatory axis that may govern these processes, providing novel insights into male fertility and potential targets for therapeutic intervention.