Ligand-receptor Interactions Research Articles

Transcriptomic and functional characterization of megakaryocytic-derived platelet-like particles: impaired aggregation and prominent anti-tumor effects

Platelet-like particles (PLPs), derived from megakaryocytic cell lines MEG-01 and K-562, are widely used as a surrogate to study platelet formation and function. We demonstrate by RNA-Seq that PLPs are transcriptionally distinct from platelets. Expression of key genes in signaling pathways promoting platelet activation/aggregation, such as the PI3K/AKT, protein kinase A, phospholipase C, and α-adrenergic and GP6 receptor pathways, was missing or under-expressed in PLPs. Functionally, PLPs do not aggregate following epinephrine, collagen, or ADP stimulation. While PLPs aggregated in response to thrombin, they did not display enhanced expression of surface markers P-selectin and activated α2bβ3, in contrast to platelets. We have previously demonstrated that platelets physically couple to MDA-PCa-2b and RC77T/E prostate cancer (PCa) cells via specific ligand-receptor interactions, leading to platelet-stimulated cell invasiveness and apoptotic resistance, and reciprocal cell-induced platelet aggregation. In contrast, PLP interactions with PCa cells inhibited both cell invasion and apoptotic resistance while failing to promote PLP aggregation. Moreover, PLPs reduced platelet-PCa cell interactions and antagonized platelet-stimulated oncogenic effects in PCa cells. RNA-Seq analysis identified candidate ligand-transmembrane protein combinations involved in anti-tumorigenic signaling of PLPs to PCa cells. Antibody neutralization of the TIMP3-MMP15 and VEGFB-FGFR1 signaling axes reversed PLP-mediated anti-invasion and apoptotic sensitization, respectively. In summary, PLPs lack many transcriptomic, molecular and functional features of platelets and possess novel anti-tumorigenic properties. These findings indicate that PLPs may have a potential therapeutic role in targeting and disrupting the oncogenic signaling between platelets and cancer cells, offering a new avenue for anti-cancer strategies.

TSPOAP1-AS1: A Novel Biomarker for the Prognosis and Therapeutic Target in Cervical Cancer.

TSPOAP1 antisense RNA 1 (TSPOAP1-AS1) is a long non-coding RNA (lncRNA) that has received widespread attention in oncology research in recent years. Its role and mechanism in some cancers have gradually been revealed. However, it is not clear what role TSPOAP1-AS1 plays in cervical cancer (CESC). In this study, bioinformatic analysis and experimental validation were carried out to investigate the relationship between TSPOAP1-AS1 and CESC. The relationships between clinical characteristics in patients with CESC, TSPOAP1-AS1 expression, prognostic factors, regulation network, and immune infiltration of TSPOAP1-AS1 were evaluated using statistics and The Cancer Genome Atlas database. Real-Time Quantitative Reverse Transcription PCR was used to test TSPOAP1-AS1, miR-17-5p, and AGFG2 expression in CESC cell lines. CESC patients exhibited markedly reduced expression of TSPOAP1-AS1. There was a significant correlation between low expression of TSPOAP1-AS1 in CESC patients and the clinical stage (p < 0.05), weight (p < 0.05), and BMI (p < 0.05). Lower expression of TSPOAP1-AS1 in patients with CESC was associated with poorer overall survival (OS) (p = 0.014) and disease-specific survival (DSS) (p = 0.030). There was also an independent correlation between high expression of TSPOAP1- AS1 (p = 0.036) and DSS in patients with CESC. TSPOAP1-AS1 was involved in the ribosome, oxidative phosphorylation, antigen processing and presentation, cell adhesion molecules (CAMs), the chemokine signaling pathway, neuroactive ligand-receptor interaction, and primary immunodeficiencies. The infiltration of immune cells and the expression of TSPOAP1-AS1 were found to be correlated. A ceRNA network of TSPOAP1-AS1/miR-17-5p/AGFG2 was constructed in CESC. In CESC, a ceRNA network involving TSPOAP1-AS1/miR-17-5p/AGFG2 was successfully established. When comparing CESC cell lines with HcerEpic, the expression of TSPOAP1-AS1 and AGFG2 decreased significantly, and the expression of miR-17-5p increased significantly. In CESC patients, low expression of TSPOAP1-AS1 was associated with poor survival and immune infiltration. It may be effective to use TSPOAP1-AS1 as a biomarker of prognosis and therapeutic target in CESC.

Mechanism of Ligusticum wallichii-Borneol in the Treatment of Cerebral Ischemic Stroke in Rats Based On Network Pharmacology, Molecular Docking, and Experimental Verification.

The pharmacodynamics, molecular biology, network pharmacology, and molecular docking mechanisms of the Ligusticum wallichii and borneol medication pair (CXBP) were investigated for ischemic stroke treatment. Effective chemical components and targets of CXBP were identified using TCMSP, ETCM, and SymMap databases, whereas ischemic stroke targets were sourced from OMIM, GeneCards, TTD, PubMed, Web of Science, CNKI, Wanfang Data, and VIP databases. Protein-protein interaction (PPI) networks were generated using the STRING database, and GO and KEGG enrichment analyses were conducted using Metascape. A "disease-pathway-target-component-drug" network was created in Cytoscape, and molecular docking was confirmed with PyMOL and AutoDock tools. Rat models of MCAO were established to evaluate neurological scores, triphenyltetrazolium chloride (TTC) staining, and Nissl staining. Key components were validated through enzyme-linked immunosorbent assay (ELISA), real-time polymerase chain reaction (PCR), and immunohistochemistry. Thirty-three active ingredients and 419 potential targets for CXBP, with key compounds including Z-6,8',7,3'-diligustilide, cedrene, (+)-alpha-funebrene, POL, dipterocarpol, oleanolic acid, 1-acetyl-beta-carboline, and erythrodiol. Critical targets included ESR1, PRKCA, and PTPN6. KEGG pathway analysis revealed 179 signaling pathways, primarily neuroactive ligand-receptor interactions, whereas GO enrichment analysis identified 2911 biological processes, 398 molecular activities, and 203 cellular components. The neurological function scores and TTC staining of the infarcted brain regions were significantly improved following CXBP intervention compared to the MCAO group. These findings were supported by Nissl staining, which demonstrated better preserved cellular morphology and a significantly higher number of Nissl vesicles in the CXBP group. ELISA analysis revealed substantial modulation in gene expression: levels of PRKCA, PTPN6, ESR1, and TNF-α changed significantly, whereas IL-1β, IL-6, and TNF-α were notably downregulated compared to the MCAO group. PCR results corroborated these findings, showing a significant decrease in PRKCA expression and marked downregulation of IL-1β, IL-6, and TNF-α, whereas ESR1 and PTPN6 levels increased significantly. Immunohistochemical analysis further confirmed these results, with the CXBP and nimodipine groups exhibiting higher ESR1 and PTPN6 expression and lower PRKCA expression compared to the MCAO group. To improve cerebral ischemia and reperfusion injury, CXBP improves ischemic stroke outcomes through key active ingredients (e.g., Z-6,8',7,3'-diligustilide, cedrene, and oleanolic acid) and targets ESR1, PRKCA, and PTPN6, modulating multiple signaling pathways to alleviate cerebral ischemia-reperfusion injury.

Specific plasma metabolite profile in intestinal Behçet’s syndrome

BackgroundIntestinal Behçet’s syndrome (IBS) has high morbidity and mortality rates with serious complications. However, there are few specific biomarkers for IBS. The purposes of this study were to investigate the distinctive metabolic changes in plasma samples between IBS patients and healthy people, active IBS and inactive IBS patients, and to identify candidate metabolic biomarkers which would be useful for diagnosing and predicting IBS.MethodsIn this study, we performed a global untargeted metabolomics approach in plasma samples from 30 IBS patients and 20 healthy subjects. P value < 0.05 and variable importance projection (VIP) values > 1 were considered to be statistically significant metabolites. Univariate receiver operating characteristic (ROC) curve analysis was plotted as a measure for assessing the clinical performance of metabolites, and area under curve (AUC) were assessed.ResultsA total of 147 differentially abundant metabolites (DAMs) were identified between IBS patients and normal control (NC) group. The potential pathways involved in the pathogenesis of IBS include linoleic acid metabolism; GABAergic synapse; biosynthesis of unsaturated fatty acids; valine, leucine and isoleucine biosynthesis; ovarian steroidogenesis; and others. In addition, a total of 103 significant metabolites were selected to distinguish active IBS from inactive IBS patients. Tyrosine metabolism, dopaminergic synapse and neuroactive ligand-receptor interaction were found to be closely related to the disease activity of IBS. Furthermore, three potential metabolites including quinate, stearidonic acid (SDA) and capric acid (CA) could significantly differ IBS patients from NC group. On the other hand, 1-methyladenosine (m1A), genipin, methylmalonic acid (MMA) and ascorbate could significantly differentiated active IBS from inactive IBS patients.ConclusionIn conclusion, this study demonstrated the characteristic plasma metabolic profiles between IBS group and NC group, as well as between active and inactive IBS patients by using an untargeted LC/MS metabolomics profiling approach. In this study, quinate, SDA and CA were identified as potential diagnostic biomarkers for IBS. Additionally, m1A, genipin, MMA and ascorbate could serve as potential biomarkers for evaluating IBS activity. These findings might provide potential valuable insights for developing therapeutic strategies to manage IBS in the future.

Characterization of cell membrane fragments containing muscle type nAChR from Tetronarce californica after preparation using high pressure homogenization.

The nicotinic acetylcholine receptor (nAChR) is a pentameric ligand-gated ion channel (pLGIC) commonly used as a model for receptors belonging to the Cys-loop superfamily. Members of pLGICs are standardly used in numerous toxicological investigations e.g., GABA and nAChR in the context of nerve agent poisoning. Organophosphorus compounds inhibit AChE, leading to accumulation of acetylcholine in the synaptic cleft and subsequently to a cholinergic crisis, in part through desensitization of nAChR. Due to the limitations of standard therapy, studies concerning functional ligand-receptor interactions of therapeutically active substances are of high importance. Therefore, we developed a novel method to obtain muscle type nAChR-containing membrane fragments from native tissue using high-pressure homogenization. The obtained microsomal fragments were characterized using Dynamic Light Scattering, laser Doppler electrophoresis and protein concentration. The microsomal membrane fragments were further purified, and the plasma membrane fraction was enriched using different density gradients. KD and BMax values were determined using a scintillation proximity assay (SPA) with [3H]epibatidine as reporter ligand. Measurement data showed that the ideal conditions to obtain microsomal membrane fragments with high pressure homogenization were four runs at 400bar. For density gradient centrifugation the under layering of the microsomal membrane fragments (bottom-up method) is to be preferred for further purification. Sucrose seems to be more efficient compared to xylitol or iodixanol density gradients. The nAChR-containing plasma membrane fractions resulting from the developed purification protocol achieve a high degree of quality and reproducibility, making them suitable to model physiological conditions. This system has the potential to be used in both bead- and filtration-based assays probing affinity parameters for ligand binding or functional experiments. The protocol can be easily modified for other LGICs or transmembrane proteins, allowing for further expansion of its use.

Zhenwu Decoction Modulates PDE3A to Alleviate Inflammation in Diabetic Nephropathy

Background: Diabetic nephropathy, a complex microvascular complication of diabetes, poses substantial health and economic challenges globally. ZWD, known for its multi-component composition, has demonstrated potential therapeutic effects in DN, yet its molecular mechanisms require further elucidation to support its clinical application. Aim: This study aimed to comprehensively investigate the active components, target molecules, and molecular mechanisms of Zhenwu Decoction (ZWD), a Traditional Chinese Medicine (TCM) formula, in the treatment of diabetic nephropathy (DN), with a focus on its anti-inflammatory effects through the modulation of the PDE3A-mediated cAMP-PKA-NFκB axis. Objective: The primary objective was to identify the active components of ZWD, their targets, and the underlying molecular pathways through which ZWD exerts its therapeutic effects on DN, specifically focusing on the modulation of inflammation in renal cells under high glucose conditions. Method: A combined approach of network pharmacology, single-cell transcriptomics, and molecular docking analyses was employed to investigate the active components, target molecules, and underlying mechanisms of ZWD in treating DN. A high-glucose-induced HK-2 cell model was established to simulate in vitro DN conditions. Cell viability was assessed using the CCK-8 assay, while quantitative real-time PCR (qPCR) and Western blotting were employed to quantify gene expression and protein levels, respectively. Results: Our analysis uncovered 141 active components in ZWD, which targeted 171 proteins involved in pathways related to hypoxia response, neuroactive ligand-receptor interaction, urotensin II signaling, and other pathways implicated in diabetes and vascular diseases. Among these, 75 overlapping genes were pinpointed as potential therapeutic targets of ZWD for DN. Protein-protein interaction networks revealed three functional clusters of targets potentially associated with oxidative stress responses, lipid metabolism, and hormonal regulation. Cell-type specific analyses showed differential expression of these targets in DN, particularly in proximal tubular epithelial cells. Notably, through molecular docking, we discovered a strong binding affinity between PDE3A and betasitosterol. In vitro experiments confirmed that ZWD extract modulated PDE3A expression, leading to the suppression of the PKA/AMPK/NF-κB axis and attenuation of high glucose-induced inflammation in HK-2 cells. Conclusion: This study identified that Zhenwu Decoction (ZWD) suppressed high glucose-induced inflammation in renal cells by inhibiting the PDE3A-mediated cAMP-PKA-NFκB axis, highlighting the potential of ZWD as an effective adjunct therapy for diabetic nephropathy and paving the way for innovative anti-inflammatory treatments.

Effects of colloids with different compositions on benzophenone-3 biotoxicity in zebrafish embryos.

The fate of the pollutants in aquatic environment is closely related to colloids, and the carrier effect of colloids on pollutants not only affects their bioaccumulation, but may also affect their toxicity. In this study, the effects of natural colloid with different components on the biological toxicity of benzophenone-3 (BP3) to zebrafish larvae (Diano rerio) were studied. BP3 caused oxidative stress damage, thyroid system disorders and neurotoxicity in zebrafish larvae. And in the co-exposure groups, the organic and black carbon mineral (BCM) colloids enhanced the organism's antioxidant system by regulating the activities of superoxide dismutase (SOD) and glutathione peroxidase (GPx), reducing the lipid peroxidation damage in larvae. BCM colloids caused the thyroid system disorders in organisms, while organic colloids exacerbated the thyroid toxicity by transporting more BP3 into organisms, inducing severe abnormal heartbeats. The BCM and organic colloids regulated the acetylcholinesterase (AChE) activity and/or 5-hydroxytryptamine (5-ht) contents by affecting the neuroactive ligand receptor interaction pathway in zebrafish larvae, significantly increasing their swimming speed in co-exposure groups under the light condition. In addition, the effects of colloid-bound and freely dissolved BP3 absorbed by organisms on their physiological and biochemical activities were different. By analyzing the relative expression of the significant differential metabolites affected by BP3 in all experimental groups, it was found that colloid-bound and freely dissolved BP3 had a synergistic effect on most of these metabolites and pathways. However, the freely dissolved BP3 interfered with the purine metabolic pathway by mediating 2-(amidino)-n1-(5-phospho-d-ribosyl)acetamidine, and the tyrosine metabolic pathway by mediating choline and uranylacetic acid, while the colloid-bound BP3 has no or inverse regulatory effects on these three metabolites. This study provided a new perspective for the biotoxicity study of the pollutants in aquatic environment, necessitating a reconsideration of the real ecological risks of emerging pollutants in the presence of natural colloids.

Exploring the potential anticancer targets and mechanistic pathways of Elsholtzia densa essential oil based on network pharmacology

This study aimed to assess the composition of Elsholtzia densa essential oil (EBE) and identify potential targets for inhibiting human hepatocellular carcinoma cell proliferation. The plants were collected from four regions: Jiuzhi, Qinghai; Ruoergai, Sichuan; Aba, Sichuan; and Jiulong, Sichuan. Four EBEs (named No. 1 to No. 4) were analyzed by gas chromatograph-mass spectrometer. EBEs significantly inhibited human hepatocellular carcinoma cells. The EBE collected from Jiuzhi exhibited the most potent inhibitory effect. Core targets identified included MAPK3, EGFR, ESR1, CASP3, PTGS2, BCL2L1, and MAPK14. Notably, the four EBEs prevented hepatocellular carcinoma cell proliferation via neuroactive ligand-receptor interactions and apoptosis pathways.

FcγR1-Expressing Cell Membrane-Coated Nanoparticle (FcγR1-CMNP) for T-Cell-Engaging Bispecific Nanoantibody Construction.

T-cell-engaging bispecific antibodies (BiTEs), which can simultaneously bind to antigens on tumor cells and T cells, show good potential in cancer immunotherapy. A practical and feasible approach for emulating BiTEs involves immobilizing two types of monoclonal antibodies (mAbs) onto a single nanoparticle; however, this approach involves complex immobilization processes and chemical reactions. To overcome these challenges, we achieved gentle antibody immobilization through receptor-ligand interactions by constructing a mAb delivery system known as Fcγ receptor 1 (FcγR1)-expressing cell membrane-coated nanoparticles (abbreviated as FcγR1-CMNPs). To validate the functionality and feasibility of this approach, we immobilized αCD3 and αCD20 onto the CMNP surface and generated a bispecific nanoantibody (termed CMNP@CD3 × CD20) for treating B-cell lymphoma. We demonstrated that CMNP@CD3 × CD20 significantly promoted the interaction between T cells and tumor cells and resulted in the potent killing of tumor cells in vitro and in vivo. This innovative antibody delivery system offers a straightforward and gentler technique for antibody delivery, potentially facilitating the development of trispecific or multispecific antibody-based immunotherapy.

Towards melanoma in situ vaccination with multiple ultra-narrow X-ray beams.

Despite the recent progress, current treatment modalities are not able to eradicate cancer. We show that Microbeam Radiotherapy (MRT), an innovative type of Spatially Fractionated Radiotherapy, can control murine melanoma by activating the host's own immune system. The beneficial effects are very pronounced in comparison to uniform radiotherapy traditionally employed in the clinic. Our results show that MRT increased antigen presentation, activating Cytotoxic T Lymphocytes (CTLs) which are essential to MRT's treatment efficacy in melanoma. Depletion of CTLs abrogated treatment response. Multiplex nucleic acid hybridization technology revealed key features of lymphocyte populations such as proliferation, differentiation, and ligand-receptor interactions. In addition, CTLs were shown to be essential for locoregional metastatic control and systemic abscopal effects confirmed by activation of antigen presenting cells and CTL trafficking in the tumour-draining lymph nodes. MRT also showed a synergistic effect with immunotherapy. Overall, MRT induces a robust antitumour immune response, acting like an in situ vaccination, which could be exploited to treat a variety of treatment-resistant malignancies.

Involvement of Tim-3 in Maternal-fetal Tolerance: A Review of Current Understanding.

As the first T cell immunoglobulin mucin (Tim) family member to be identified, Tim-3 is a powerful immune checkpoint that functions in immunoregulation and induction of tolerance. Conventionally, Tim-3 is considered to play a role in adaptive immunity, especially in helper T cell-mediated immune responses. As researches progress, Tim-3 has been detected in a wider range of cell types, modulating cell function through ligand-receptor interactions and other pathways. Strikingly, Tim-3 plays a pivotal role in maternal-fetal tolerance by regulating immune cell functions and orchestrating the maternal-fetal cross-talk. In this review, we elaborate on the involvement of Tim-3 in immunology, with a focus on its participation in maternal-fetal tolerance to provide new insights into immunoregulation during pregnancy. Our work will be helpful in further understanding the pathogenesis of pregnancy-related diseases and will inspire new strategies for their diagnosis and treatment.

Effects of cage size on the natural behavior, serum parameters, production performance and hypothalamic mRNA expression profiles of laying hens

The objective of this study was to evaluate the effects of cage size on the natural behavior, serum biochemistry, production performance and hypothalamic transcriptome profiles of laying hens. A total of 360 79-week-old hens were selected and randomly assigned to three groups (with five replicates each) with different cage sizes: large cages (LCs), medium cages (MCs), and small cages (SCs). The stocking density remained consistent across all groups throughout the experimental period. Behavioral expression was evaluated through observation of only six birds per group on a weekly basis. Compared to the SC group, the average frequencies of walking, wing-flapping, feather-pecking and drinking significantly increased in the LC and MC groups (P <0.05), whereas the average frequency of stereotypic behavior significantly decreased (P <0.05), indicating that cage size has positive effects on natural behavior. The average daily step count (DSC) was in the order LC > MC > SC (P <0.05), which suggested an improvement in exercise ability in larger cages. The serum levels of MDA, CORT, AST and T-CH were significantly lower in the LC and MC groups than in the SC group (P <0.05), whereas the serum activities of SOD and GSH-Px were significantly greater (P <0.05). Additionally, compared to the SC group, the laying rates were increased by 1.61% and 2.20% (P <0.05), and the feed conversion ratio (FCR) were improved by 1.35% and 2.25%, respectively (P <0.05) in the LC and MC groups, respectively. In total, 437 and 81 differentially expressed genes (DEGs) were identified in the hypothalamus of birds between the LC and SC groups and the MC and SC groups, respectively. A majority of these DEGs were involved primarily in neuroactive ligand‒receptor interactions, focal adhesion and calcium signaling pathways, which are associated with the regulation of behavioral patterns, the stress response and follicular development. Our results suggest that an appropriate increase in cage size is beneficial to the natural behavior expression, health status and production performance of laying hens.

Integrated Network Pharmacology and In-silico Approaches to Decipher the Pharmacological Mechanism of Dioscorea septemloba Thunb in Treating Gout and Its Complications

Background: Dioscorea septemloba Thunb. (DST) has demonstrated therapeutic potential in the treatment of gout and its associated complications. However, the underlying mechanisms of DST’s pharmacological activity remain unclear. This study aims to investigate the pharmacological substances and network regulatory mechanisms of DST in treating gout and its complications using network pharmacology. Methods: According to ultra-high performance liquid chromatography coupled with hybrid quadrupole-Orbitrap mass spectrometry (UPLC-Q-Exactive Orbitrap-MS) data and Lipinski’s rule of five, 24 bioactive phytochemicals from DST were identified. The targets of gout were retrieved from Gene Expression Omnibus (GEO), GeneCards, and DisGeNET databases, followed by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes pathway (KEGG pathway) enrichment analysis. The Cytoscape network analysis was used to identify the primary pathological pathways and key targets. Finally, LeDock was used for molecular docking to verify the active components of DST and their core target proteins. Results: DST contains several core active ingredients, such as tetrahydroimidazo[1,2-a]pyridine- 2,5-dione, diosgenin, beta-sitosterol, dioscorol B, montroumarin and 9,10-dihydro-5,7- dimethoxy-3,4-phenanthrenediol. Moreover, these active components were found to strongly bind to the key targets for treating gout and its complications, including HSP90AA1, STAT3, PTGS2, PPARG, MTOR, HIF1A, MMP9, ESR1, and TLR4. As a result, DST alleviates gout and its complications by inhibiting xanthine dehydrogenase (XDH) to reduce uric acid levels and regulating the HIF-1α, EZH2/STAT3, and COX-2/PPAR-γ pathways to reduce inflammation. Additionally, it also plays an analgesic role by regulating the neuroactive ligand-receptor interaction pathway and calcium ion signaling pathway. Conclusion: This study has provided insights into the underlying mechanisms of DST in the treatment of gout and its complications, which could serve as a scientific foundation for its clinical translation.

Unprecedented carbonic anhydrase inhibition mechanism: Targeting histidine 64 side chain through a halogen bond.

2,2'-Thio-bis(4,6-dichlorophenol), namely bithionol, is a small molecule endowed with a multifaceted bioactivity. Its peculiar polychlorinated phenolic structure makes it a suitable candidate to explore its potentialities in establishing interaction patterns with enzymes of MedChem interest, such as the human carbonic anhydrase (hCA) metalloenzymes. Herein, bithionol was tested on a panel of specific hCAs through the stopped-flow technique, showing a promising micromolar inhibitory activity for the hCA II isoform. X-ray crystallographic studies revealed an unprecedented halogen-bond interaction between one chlorine of bithionol and the N3(ε) atom of the hCA II catalytically active histidine residue, His64. Then, quantum mechanics calculations based on the fragment molecular orbital method allowed us to estimate the strength of this bond (~2.9 kcal/mol) and highlighted the contribution of a rich hydrophobic interaction network within the isoenzyme. Interestingly, the compound inactivity against the hCA III isoform, characterized by His64Lys and Leu198Phe mutations, supported the key role played by halogen bonding in the enzyme affinity. This finding might pave the way for the development of a new class of hCA inhibitors characterized by such chemical features, with the halogen bond being a key ligand-receptor interaction.

Diagnostic and Prognostic Potential of FBXO8 Expression in Kidney Renal Clear Cell Carcinoma and Its Regulation of Renal Adenocarcinoma Cells

BackgroundThe F-box protein 8 Gene (FBXO8) has been shown to suppress invasion and metastasis in various cancer types. Recurrence and drug resistance pose significant challenges in renal cell carcinoma (RCC). Identifying novel biomarkers is crucial for addressing these issues. MethodsData on RNA sequencing and patient survival for KIRC was obtained from The Cancer Genome Atlas (TCGA), UALCAN, and Gene Expression Omnibus (GEO) databases. We confirmed FBXO8 gene expression and its impact on survival. Clinical characteristics were classified, and FBXO8 expression differences among various categories were observed. We conducted biofunctional predictions and analyzed the tumor microenvironment (TME), immune cell infiltration, and immune checkpoints in relation to FBXO8 expression. FBXO8 was overexpressed using a plasmid, and we assessed Kidney renal clear cell carcinoma (KIRC) cell proliferation, migration, and apoptosis through CCK8, wound healing tests, and western blot analysis. ResultsOur findings revealed decreased FBXO8 expression in KIRC, with patients exhibiting low FBXO8 expression experiencing shorter survival times. The low expression group showed elevated TME immune and estimate scores. Biofunctional analyses indicated that FBXO8 expression was notably linked to drug metabolism cytochrome P450, nutrition disease, receptor-ligand activity, and neuroactive ligand-receptor interaction. Furthermore, we discovered significant correlations between FBXO8 expression and immune cell infiltration, as well as checkpoints such as CD274. Overexpression (OE) of FBXO8 led to a marked reduction in cell proliferation and migration, along with increased apoptosis, as evidenced by apoptosis-related protein expression. ConclusionThis study demonstrates that FBXO8 serves as a biomarker for KIRC and plays a role in regulating cell proliferation, migration, and apoptosis.

NLRP3 Inhibitor Alleviates Glycemic Variability-Induced Cognitive Impairment in Aged Rats with Type 2 Diabetes Mellitus

Glycemic variability (GV) markedly exacerbates cognitive impairment in elderly patients with type 2 diabetes mellitus (T2DM), in part through chronic inflammation. This study investigated the therapeutic efficacy of the NLRP3 inflammasome inhibitor MCC950 in mitigating GV-induced cognitive impairment in an aged rat model of T2DM. Aged Sprague-Dawley rats with induced T2DM were subjected to GV conditions, and the effects of MCC950 were evaluated through measurement of body weight, blood glucose, lipid profiles, insulin level, inflammatory markers, and cognitive function. Transcriptomic analysis was performed on the hippocampus and prefrontal cortex. Treatment with MCC950 significantly alleviated weight loss and hyperglycemia in the GV group compared with the control group. MCC950 also reduced the levels of cholesterol, triglycerides, and pro-inflammatory markers (interleukin-1β (IL-1β) and interleukin-18 (IL-18)). Most notably, MCC950 improved spatial learning and memory retention in the GV group. Immunohistochemical analysis indicated a reduction in inflammasome activation and an increase in the expression level of the neuronal marker NeuN in the hippocampus. Transcriptomic analysis revealed that MCC950 altered neuroactive ligand-receptor interaction pathways in the hippocampus and influenced receptor binding and cell adhesion processes in the prefrontal cortex. These findings validated the efficacy of NLRP3 inhibitor in mitigating GV-induced cognitive impairment in elderly rats with T2DM and provided the basis for subsequent clinical studies exploring the broader potential of NLRP3-targeted interventions in addressing diabetes-associated cognitive impairment.

Effects of early weaning on the development of gastrointestinal tract, rumen microbiota and host gene regulation in goat kids

Early weaning (EW) could shorten ewes’ reproductive cycles and promote rumen development in goat kids. During the transition from breastfeeding to a solid diet, significant changes occur in rumen development. However, few studies have focused on goat kids, which are more sensitive to stress than lambs. Here, we assessed the effects of EW on hematological parameters, rumen microbiota, and host gene expression in Chengdu Brown goat kids. The EW kids (weaned at day 42) did not display differences in growth performance and blood biochemical parameters compared to the control group (Control group, CON). However, these younger kids had higher levels of butyric acid than the CON group. EW altered the composition of rumen microbiota and host gene expression, with genes such as RNF183, SCD, SAMD8, MME, and TULP1 being up-regulated, all of which are related to protein generation or binding. Conversely, ADRA1A, GLI1, GRIA4, and CACNA1B were down-regulated, involving neuroactive ligand-receptor interaction, MAPK, and cAMP signaling pathways. Furthermore, Christensenellaceae_R_7_group and Prevotella showed significant correlations with the expressions of differently expressed genes DMGDH and FABP. These results could be beneficial to understanding the interactions among indicators in goat kids around weaning and facilitating management of weaning practices in intensive production systems.

Cell-type-specific regulators landscape and regulatory mechanisms underlying pyroptosis in uterine corpus endometrial carcinoma.

Background: Endometrial cancer (UCEC) has a significant detrimental effect on patient quality of life. Although pyroptosis-related genes have been reported to contribute to tumor pathogenesis, the specific mechanism of pyroptosis in patients with UCEC remains elusive. Methods: We provide an overview of the landscape of pyroptosis-related genes in UCEC tissues through single-cell RNA sequencing (scRNA-Seq) datasets from the tissues of UCEC of 6089 cells. In addition, pyroptosis-related gene expression pattern was verified based on the RNA-Seq datasets, and observation of abnormal pathological characteristics of UCEC tissue. Results: The pyroptosis-related gene IL-6 is specifically upregulated in epithelial cells and dysregulates cell population proliferation and enhances apoptosis. The upregulation of BAX and TNF expression in macrophages induces infiltration of aberrantly activated macrophages, which display dysfunctional differentiation in tumor tissues, altered immune responses, and activation of the tumor necrosis factor (TNF) pathway in UCEC macrophages. In addition, dysregulation of pyroptosis-related genes induces aberrant cell-cell communication in tumor tissues and mediates ligand-receptor interactions between various cell types in UCEC via the TNF signaling pathway to promote cancer progression. Quantitative real-time (PCR) and immunohistochemistry were used for the in vitro experimental validation. Conclusion: Pyroptosis-related genes can serve as biomarkers for UCEC, playing a role in early disease diagnosis, helping to predict patient prognosis, and guiding the selection of personalized treatment options.

Comprehensive analysis of heterogeneity and cell-cell interactions in Crohn's disease reveals novel location-specific insights.

In Crohn's disease (CD), lesions are mainly distributed in a segmental manner, with the primary sites of involvement being the ileum and colon. Heterogeneity in colon and ileum results in location-specific clinical presentations and therapeutic responses. Mucosal healing tends to be more readily and quickly achieved in the colon than in the ileum, where lesions are more likely to develop into complex behaviors. The heterogeneity of colon and ileum in CD, which is essential for tailored therapeutic approaches, has not yet been systematically illustrated. CD presents with unique intestinal lesions, mainly impacting the terminal ileum and colon. It is essential to comprehend the diversity in pathogenesis and treatment response among various segments. We conducted comparative single-cell RNA sequencing analysis in treatment-naïve CD patients, concentrating on the colon and ileum. A novel subset of epithelial cells expressing high levels of DUOX2 and DUOXA2 (DUOX2-epi) was discovered. This DUOX2-epi subcluster predominantly distributed in the tip epithelium of the inflamed colon, potentially in response to microbial infection, as evidenced by the significant enrichment of inflammatory and microbial response pathways. The colonic and ileal DUOX2-epi subsets trigger inflammatory responses through distinct mechanisms. The colonic DUOX2-epi primarily affects monocytes via the SAA1-FPR2 ligand-receptor interaction, whereas the ileal DUOX2-epi directly interacts with regulate T cells through the CXCL16-CXCR6 ligand-receptor pair. Moreover, the cell-cell communication networks involving DUOX2-epi in the colon and ileum can help predict the location-specific effects of biological therapies. This study delves into the heterogeneity within the ileum and colon of Crohn's disease at the single-cell level, identifying a new epithelial subset DUOX2-epi. Predictive gene modules tailored to different locations for biological therapies are developed as well, based on the cell-cell communication network modulated by DUOX2-epi.

Probing the Histamine H1 Receptor Binding Site to Explore Ligand Binding Kinetics.

Analysis of structure-kinetic relationships (SKR) can contribute to an improved understanding of receptor-ligand interactions. Here, fragment 1 (4-(2-benzylphenoxy)-1-methylpiperidine) was used in different fragment growing approaches to mimic the putative binding mode of the long residence time (RT) ligands olopatadine, acrivastine, and levocetirizine at the histamine H1 receptor (H1R). SKR analyses reveal that introduction of a carboxylic acid moiety can increase RT at H1R up to 11-fold. Ligand efficiency (LE) decreases upon the introduction of the negatively charged group, whereas kinetic efficiency (KE) increases up to 8.5-fold. The olopatadine/acrivastine mimics give up to 15-fold differences in the RT, while the levocetirizine mimics afford similar RTs with only a 3-fold difference. Therefore, the levocetirizine mimics are less sensitive to structural changes. This study illustrates that for H1R, there are several ways to increase RT but the different strategies differ significantly in SKR.