IL-6 Gene Research Articles

Probiotic intervention and exercise mitigate inflammation and histopathological alterations in the liver of wistar rats on a high-fat diet.

Controlling intestinal risk factors by consuming probiotics and modifying lifestyle with exercise modulates dietary damage. The aim of the present study was to investigate the effect of 6weeks of aerobic exercise training and probiotic consumption on the expression of inflammatory genes and histopathological changes in the liver of rats with a high-fat diet model. In this study, 40 male Wistar rats were divided into 5 groups: healthy control, high-fat diet (HFD), HFD with exercise (HFD + Exe), HFD with probiotic consumption (HFD + Prob), and HFD + Exe + Prob. Animals in the HFD group were first exposed to a special diet and after confirming liver tissue damage, they entered the main protocol. Animals in the exercise group performed aerobic exercise on a rodent treadmill for 6weeks, 5days a week. Animals in the probiotic group also received Lactobacillus bifidus by oral gavage after exercise. Finally, intestinal and liver tissue were removed and examined for histological and cellular examination. Based on the results, HFD caused tissue damage and fat infiltration in both intestinal and liver tissue. Also, inflammatory factors (IL-6 and IL-1β genes) in the liver tissue of this group increased significantly compared to the control group (p < 0.05). In contrast, probiotic intervention and aerobic exercise caused a significant decrease in IL-6 and IL-1β genes compared to the HFD group (p < 0.05). The use of probiotic Lactobacillus bifidus along with exercise can neutralize inflammatory damage caused by a high-fat diet in liver tissue. However, further studies are needed in this field.

Ezh2 Shapes T Cell Plasticity to Drive Atherosclerosis.

The activation and polarization of T cells play a crucial role in atherosclerosis and dictate athero-inflammation. The epigenetic enzyme EZH2 (enhancer of zeste homolog 2) mediates the H3K27me3 (trimethylation of histone H3 lysine 27) and is pivotal in controlling T cell responses. To detail the role of T cell EZH2 in atherosclerosis, we used human carotid endarterectomy specimens to reveal plaque expression and geography of EZH2. Atherosclerosis-prone Apoe (apolipoprotein E)-deficient mice with CD (cluster of differentiation) 4+ or CD8+ T cell-specific Ezh2 deletion (Ezh2cd4-knockout [KO], Ezh2cd8-KO) were analyzed to unravel the role of T cell Ezh2 in atherosclerosis and T cell-associated immune status. EZH2 expression is elevated in advanced human atherosclerotic plaques and primarily expressed in the T cell nucleus, suggesting the importance of canonical EZH2 function in atherosclerosis. Ezh2cd4-KO, but not Ezh2cd8-KO, mice showed reduced atherosclerosis with fewer advanced plaques, which contained less collagen and macrophages, indicating that Ezh2 in CD4+ T cells drives atherosclerosis. In-depth analysis of CD4+ T cells of Ezh2cd4-KO mice revealed that absence of Ezh2 results in a type 2 immune response with increased Il-4 (interleukin 4) gene and protein expression in the aorta and lymphoid organs. In vitro, Ezh2-deficient T cells polarized macrophages toward an anti-inflammatory phenotype. Single-cell RNA-sequencing of splenic T cells revealed that Ezh2 deficiency reduced naive, Ccl5+ (C-C motif chemokine ligand 5) and regulatory T cell populations and increased the frequencies of memory T cells and invariant natural killer T (iNKT) cells. Flow cytometric analysis identified a shift toward Th2 (type 2 T helper) effector CD4+ T cells in Ezh2cd4-KO mice and confirmed a profound increase in splenic iNKT cells with increased expression of Plzf (promyelocytic leukemia zinc finger), which is the characteristic marker of the iNKT2 subset. Likewise, Zbtb16 ([zinc finger and BTB domain containing 16], a Plzf-encoding gene) transcripts were elevated in the aorta of Ezh2cd4-KO mice, suggesting an accumulation of iNKT2 cells in the plaque. H3K27me3-chromatin immunoprecipitation followed by quantitative polymerase chain reaction showed that T cell-Ezh2 regulates the transcription of the Il-4 and Zbtb16 genes. Our study uncovers the importance of T cell EZH2 in human and mouse atherosclerosis. Inhibition of Ezh2 in CD4+ T cells drives type 2 immune responses, resulting in an accumulation of iNKT2 and Th2 cells, memory T cells and anti-inflammatory macrophages that limit the progression of atherosclerosis.

Common pathogenetic mechanisms of inflammatory bowel diseases and atherosclerosis: a focus on cytokines

Objective: To analyze literature data and compare the common pathways of inflammation in inflammatory bowel diseases (IBD) and atherosclerosis with focus on the effects of proinflammatory cytokines on both pathologies, as well as from the perspective of potential role of gene polymorphism of proinflammatory cytokines in the pathophysiology of atherosclerosis in IBD patients. Main provisions: In IBD, a serum cytokine profile initiates and supports the chronic inflammation. The main immunological mechanisms in both IBD and atherosclerosis are mediated by hyperproduction of proinflammatory cytokines, such as tumor necrosis factor α (TNF-α), interleukins (IL) IL-1β, IL-6, IL-8, IL-12, IL-23, IL-17, and relative insufficiency of anti-inflammatory IL-4 and IL-10, with significant increase in pro-inflammatory cytokines. An association has been established between the polymorphisms of TNF-α (rs1800629), IL-8 (rs117518778, rs8057084), IL-10 (rs3024505, rs1800896), IL-12 (rs6887695, rs10045431), IL-23 (rs11209026A) candidate genes and the development of ulcerative colitis (UC). The polymorphisms of the TNF-α (rs1800629), IL-8 (rs117518778, rs8057084), IL-1β (rs16944), IL-17A (rs2275913), IL-4 (rs2243250), IL-23 (rs6682925T/C) genes are associated with a high risk of atherosclerosis. Protein tyrosine phosphatase, receptor type, C (PTPRC) was identified as the hub crosstalk gene for the comorbidity of UC and atherosclerosis. The effects of cytokine genes polymorphisms as key targets of the pathogenetically oriented IBD therapy on the development of atherosclerosis in these patients remain a poorly investigated question. Conclusion: IBD and atherosclerosis are mediated by the shared mechanisms of enhanced synthesis of proinflammatory cytokines, as well as polymorphisms of the candidate genes. Studies on the polymorphism of proinflammatory cytokines genes and small molecules in patients with UC, as well as the association of these polymorphisms with the development of atherosclerosis would open up new possibilities for prediction of cardiovascular diseases in these patients, development of preventive measures, and for repositioning of biological therapy for the prevention and treatment of atherosclerosis.

Individual Genomic Loci, Transcript Level and Serum Profile of Immune, Antioxidant and Hormonal Markers Associated with Sheep Arthritis

Arthritis is a leading cause of economic loss in livestock farming including sheep. This study examined the changes in gene expression, antioxidants, pro-inflammatory cytokines, acute-phase proteins (APPs), hormonal assays and iron profiles linked to sheep arthritis, as well as the diagnostic utility of these markers. Blood samples were obtained from 30 apparently healthy rams and 30 rams with arthritis for gene expression and biochemical analyses. Gene expression intensities were much higher in the arthritis-affected rams than in the healthy ones for the genes IL-1α, IL-1β, IL-6, IL-10, TNFα, NCF4, NFKB, TMED, FCAMR, iNOS and COX18. The SOD3, CAT, GPX and ATOX1 genes were expressed at substantially lower levels in arthritis-affected rams. Disparities in the nucleotide sequence variants for the amplified DNA bases linked to arthritis for the studied genes were found in the PCR-DNA sequence verdicts of the affected and healthy rams. Immunological, acute-phase protein (APP), antioxidant, hormonal and iron profiles were estimated in both groups and statistically analyzed. The arthritic group in relation to the healthy one showed a significant (p &lt; 0.05) increase in pro-inflammatory cytokines, APPs, free radicals, immunoglobulins, cortisol, GH, TSH, ferritin, TIBC and UIBC and a significant (p ˂ 0.05) decrease in anti-inflammatory cytokines, antioxidants, complements, insulin, T3, T4, SI, and Tf and Tf sat.% serum levels. The estimated pro-inflammatory cytokines and APPs achieved high values of sensitivity and specificity, positive predictive values (PPVs), negative predictive values (NPVs), a high accuracy rate and a moderate likelihood ratio (LR). The study concluded that ovine arthritis stimulates innate and humeral immunity, resulting in prominent alterations in gene expression, pro-inflammatory cytokines, APP assays and antioxidant profiles, which could be valuable indicators of sheep arthritis.

Unveiling the role of ACTL6A in uveal melanoma metastasis and immune microenvironment.

To predict and evaluate the possible mechanisms and clinical value of ACTL6A in the prognosis and development of UM. Bioinformatics analyze the relationship between ACTL6A and immunity in UM, which derived from TCGA, Gene Expression Omnibus (GEO) databases. Tumor-infltrated immune cells were demonstrated using QUANTISEQ and MCP-counter. Furthermore, scRNA-seq was used to detect ACTL6A expression, distribution, immune infiltration and revealing the gene expression profile of UM. The expression of ACTL6A was lower in UM compared with pantumor in TCGA databases. Kaplan-Meier analysis revealed that downregulated ACTL6A was associated with poor OS, and ACTL6A was associated with cancer stem cells (CSCs) and immune infiltration. Moreover, ACTL6A might act as a chemotherapy resistance gene and closely relate- to epithelial-mesenchymal transition. Analysis in 8 GSE databases showed that IL13, TPTE, IL17B and CCL22 genes were significantly overexpressed in metastatic UM. Furthermore, the single-cell transcriptomic profling identified a new cell cluster - as a unique type of immune cell, which associating with malignant cell heterogeneity and complexity, and further revealing that the metastasis of UM is mainly associated with CD4 Tconv, B , CD8 Tex, and Plasma cells. Downregulated ACTL6A acts as a risk factor for poor prognosis in UM, which implies as an potential prognostic marker for independent targeted immunotherapy.

WCN25-2101 Association of IL-10 gene promoter polymorphisms (-592A/C; -819C/T) with diabetic nephropathy in type 2 diabetes mellitus among the South Indian population

WCN25-2101 Association of IL-10 gene promoter polymorphisms (-592A/C; -819C/T) with diabetic nephropathy in type 2 diabetes mellitus among the South Indian population

RBT-1, a "preconditioning" agent, mitigates syndecan-1 shedding in patients undergoing "on pump" cardiac surgery and following experimental AKI.

During systemic stress, syndecan-1 (SDC-1) shedding into plasma results, implying endothelial damage. RBT-1, a "preconditioning" agent, has been shown to mitigate postoperative complications following cardiac surgeries. This study assessed whether RBT-1 preconditioning attenuated SDC-1 shedding in these patients, implying a vascular protective effect. Patients (n, 112) were randomized to receive low-dose RBT-1, high-dose RBT-1, or placebo 24-48 h prior to surgery. Plasma samples were obtained before and 2 days postsurgery and assayed for SDC-1 (ELISA). To gain further insights, male CD-1 mice were subjected to acute renal injuries, and RBT-1's impact on plasma SDC-1 increases, vascular/aortic stress responses (NGAL/KIM-1/IL-6 gene induction), and two vascular cytoprotective pathways (Nrf2; ferritin) were assessed. Baseline plasma SDC-1 levels did not differ between patient groups. The placebo group developed an approximate 50% plasma SDC-1 (ng/mL) increase (p, 0.012). Conversely, no significant SDC-1 increases were seen in the RBT-1 treatment groups. Experimental injury markedly increased plasma SDC-1 concentrations, and these were significantly blunted by RBT-1 preconditioning. RBT-1 also mitigated AKI-induced aortic NGAL/KIM-1/IL-6 mRNA increases, activated aortic Nrf2, and increased vascular ferritin levels. RBT-1 preconditioning diminishes SDC-1 release and upregulates vascular ferritin and Nrf2. Hence, RBT-1 preconditioning can confer select vasoprotective effects.

Association Between Active DNA Demethylation and Liver Fibrosis in Individuals with Metabolic-Associated Steatotic Liver Disease (MASLD)

Metabolic-associated steatotic liver disease (MASLD) represents the most common chronic hepatopathy worldwide and an independent risk factor for cardiovascular disease and mortality, particularly when liver fibrosis occurs. Epigenetic alterations, such as DNA methylation, may influence MASLD susceptibility and progression; yet mechanisms underlying this process are limited. This study aimed to investigate whether active DNA demethylation in peripheral blood mononuclear cells (PBMCs) from individuals with MASLD, alongside the methylation and mRNA levels of inflammation- and fibrosis-related candidate genes, is associated with liver fibrosis. For this study, global demethylation intermediates (5-hydroxymethylcytosine [5hmC], 5-formylcytosine [5fC]) were quantified in PBMCs from 89 individuals with/without MASLD using ELISA. Site-specific DNA methylation of SOCS3, SREBF1, and TXNIP was analyzed by mass spectrometry-based bisulfite sequencing; mRNA expression was assessed via RT-PCR. Individuals with MASLD and moderate-to-high fibrosis risk (estimated by the fibrosis non-alcoholic steatohepatitis (NASH) index, FNI) progressively exhibited greater global 5hmC and 5fC levels. Higher FNI was associated with reduced methylation of the SOCS3 gene and increased mRNA expression of the SOCS3, TXNIP, IL-6, and MCP-1 genes. In conclusion, elevated fibrosis risk in MASLD is associated with active global DNA demethylation, as well as differential methylation and expression patterns of genes, which are key regulators of inflammation and fibrosis. These epigenetic alterations in PBMCs may mirror DNA methylation changes in the liver, which may potentially contribute to liver fibrogenesis and represent novel biomarkers for MASLD progression toward fibrosis.

Polymorphism of genes of the cytokine system in pathogenesis of the vibration disease among workers of mining and processing enterprises

Introduction. Vibration disease (VD) is an occupational disease that develops with prolonged exposure to industrial vibration of different frequencies. VD is one of the most common occupational pathologies. This disease is characterized by the complexity of pathogenetic mechanisms, polymorphism of symptoms, chronic course, therapeutic resistance, and often leads to inability to work and disability in patients. As a rule, VD is observed to be associated with changes in the cardiovascular, nervous, and musculoskeletal system and metabolic processes. Materials and Methods. During the study, seventy three people with various forms of VD were examined. The control group consisted of 73 people who never exposed to vibration in their occupational activities. Results. The distribution of polymorphic loci rs16944, rs361525, rs1800795 was studied. Statistical analysis was performed using IBM SPSS Statistica v.21 and Microsoft Excel software packages. rs16944 is not associated with the development of VD, but is associated with the development of cerebrovascular disease in VD patients. rs1800795 of the IL6 gene also did not show an association with the development of VD, but an association of the *C/*C genotype and the *C allele with peripheral angiodystonic syndrome was demonstrated. The rs361525 locus of the TNFA gene also showed an association with peripheral angiodystonic syndrome and is also associated with the development of hypertension in VD patients. The limitations of this work include the small sample size and the fact that the study participants from the control sample work at another enterprise, which does not allow both excluding the influence of other environmental factors and considering them as a reference population. Conclusion. Thus, we were unable to identify a direct association of the studied genotypes with VD, but the association with peripheral angiodystonic syndrome suggests that these polymorphisms to influence on the pathogenesis of VD.

Tumor cells upregulate neurotransmitter GABA in the choroid plexus through STAT6-Bestrophin1 signaling, promoting leptomeningeal dissemination.

Leptomeningeal dissemination (LMD) occurs when tumor cells interact with choroid plexus epithelium (CPE) to gain access to cerebrospinal fluid (CSF) in the brain's meninges and ventricular system. This disease is particularly devastating for patients due to our limited understanding and few therapeutic options. The leptomeningeal CSF is a nutritionally deprived microenvironment for tumor cells. Despite this, LMD tumor cells survive by taking up and metabolizing the neurotransmitter GABA from the CSF. However, we currently lack evidence on how CSF-GABA levels are altered and how tumor cells communicate with the choroid plexus epithelium to increase GABA levels in the LMD microenvironment. Herein, we examined the interactions between CPEs and tumor cells that make CSF more hospitable to LMD growth. We utilized primary choroid plexus, breast cancer cell and patient-derived breast and lung to brain metastatic cells along with in-vivo modeling along with STAT6 inhibitor and IL13 gene knockdown. We show breast and lung cancer cells derived-IL13 activates STAT6 signaling in choroid plexus. Subsequently, choroid plexus upregulates GABA-synthesizing enzyme GAD67 and GABA-permeable channel Bestrophin-1, all leading to elevated neurotransmitter GABA levels in the cerebrospinal fluid. Moreover, we show a significant reduction of Bestrophin1 in choroid plexus when tumor-derived IL13 is knocked down or when treated with brain permeable STAT6 specific inhibitor AS1517499, leading to increased survival. Overall, these findings reveal a novel STAT6-Bestrophin1-GABA axis in choroid plexus and its therapeutic targeting can lead to favorable outcomes in leptomeningeal disease.

IL-33 signaling is dispensable for the IL-10-induced enhancement of mast cell responses during food allergy

BackgroundThe IL-33/ST2 axis plays a pivotal role in the development of IgE-mediated mast cell (MC) responses during food allergy. We recently demonstrated that the pleiotropic cytokine, IL-10, not only exerts proinflammatory effects on IgE-mediated MC activation, but also promotes IL-33-induced MC responses. However, whether IL-33 is necessary for IL-10’s proinflammatory effects has not been examined.MethodsTo therefore determine the role of the IL-33/ST2 axis in this pathway, we assessed the effects of IL-10 on IgE-mediated MC activation and food allergy development in wild-type (WT) and ST2-/- mice.ResultsIL-10 stimulation significantly enhanced IL-33 gene expression, ST2 receptor expression, cytokine production, mMCP-1 secretion, and proliferation in IgE and antigen-activated bone marrow-derived MCs (BMMCs) from WT mice. ST2-/- BMMCs exhibited reduced cytokine secretion in response to IgE-dependent activation. However, IL-10 enhanced cytokine production, mMCP-1 secretion, and proliferation in these cells as well. To further assess the role of IL-10, food allergy was induced in WT and ST2-/- mice subjected to antibody-mediated IL-10 depletion. IL-10-depleted WT mice exhibited a significant attenuation in MC-mediated responses to OVA challenge. While ST2-/- mice also exhibited a profound suppression of MC responses, IL-10 depletion had no additional effects. However, ST2-/-/IL-10-/- mice exhibited further decreases in OVA-IgE and antigen-specific MC activation compared to ST2-/- mice.ConclusionOur data demonstrates that IL-10 can enhance MC responses in both WT and ST2-/- mice, further corroborating its proinflammatory effects on MCs and suggesting that they are not regulated by IL-33 signaling.

Distinct single cell transcriptional profile in CD4+ T-lymphocytes among obese children with asthma.

Introduction: Obesity is a risk factor for asthma morbidity, associated with less responsiveness to inhaled corticosteroids. CD4+ T-cells are central to the immunology of asthma and may contribute to the unique obese asthma phenotype. We sought to characterize the single cell CD4+ Transcriptional profile differences in obese children with asthma compared to normal weight children with asthma. Methods: Eight normal weight and obese participants with asthma were clinically phenotyped and matched based on asthma control. Peripheral blood (PB) CD4+ T-cells were sorted, and single cell RNA sequencing was conducted. Cell clusters were identified by canonical gene expression, differential gene expression and reactome pathway analysis was applied. The obese PB bulk transcriptomic signature from the U-BIOPRED pediatric cohort was assessed in our cohort as well. Results: Obese children with asthma have a distinct CD4+ transcriptional profile with differential gene expression. There were more activated protein tyrosine phosphate receptor type C (PTPRC)high cells and less PTPRClow in the obese children. Obese children had higher enrichment of the neutrophil degranulation, interleukin-7 (IL-7) receptor and IL-7-related janus kinase-signal transducer and activator of transcription signaling pathways. Genes previously associated with more severe asthma, IL-32, FKBP5 gene expression, IL-6 and Rho transcriptional signaling, were also enriched in obese children with asthma. Discussion: Our findings shed insight into the molecular mechanisms underpinning more severe and steroid-resistant asthma among obese children. Further investigation is needed to identify potential new therapeutic targets for this group.

Rhanterium Epapposum Essential Oil and Its Primary Compounds Control Infection, Inflammation, and Serum Electrolyte Imbalance in Mice with Giardiasis.

The present experimental study seeks to evaluate the in vitro and in vivo effects, as well as the potential mechanisms of action, of Rhanterium epapposum essential oil (REE) and its main constituents against Giardia lamblia infection. The analysis of REE was performed using the Gas Chromatography-Mass Spectrometry (GC-MS) detector. The in vitro effects of REE and its main constituents on viability of G. lamblia cysts and the human intestinal epithelial cell line, as well as their effects on plasma membrane permeability, the reactive oxygen species (ROS) generation, and trophozoite adherence were assessed by MTT assay. Effects of oral administration of REE and its main constituents at doses 10 and 20mg/kg on the number of Giardia cysts, the serum level of electrolytes of sodium (Na+) and potassium (K+), as well as the gene expression of inflammatory genes of TNF-α, IL-1β, IL-10, NF-κB p65, and Toll-like receptor 4 (TLR4)) were assessed. The 50% inhibitory concentrations (IC50) value of REE, MC, CP, LN and MTZ against G. lamblia cysts was 26.7, 32.1, 35.3, 45.1, and 42.1µg/mL, respectively; indicated that REE, MC, and CP displayed high antigiardial effects in comparison with MTZ. The obtained 50% cytotoxic concentration (CC50) value of REE, MC, CP, LN, and MTZ on HIEC-6 cells was 279.1, 363.4, 394.2, 478.3, and 422.1µg/mL, respectively. REE and its main compounds significantly (p < 0.001) increased the plasma membrane permeability and ROS generation in Giardia trophozoites, while, reduced the Giardia trophozoites adherence. In vivo, REE, MC, CP, and LN mainly at the dose of 20mg/kg considerably reduced the number and viability of Giardia cysts, modulated the serum level of electrolytes Na and K in the infected mice compared with the mice treated with MTZ (P < 0.01); whereas inhibited the inflammation cytokines in the infected mice compared with the mice treated with MTZ (p < 0.001). The current investigation showed that R. epapposum essential oil and its main compounds controlled the Giardia infection in mice by inhibiting inflammation, and serum electrolyte imbalance. After confirmation in further studies, these compounds may be considered for development as a novel therapeutic option for the treatment of giardiasis.

Exploring the therapeutic effect of human recombinant IL11 on lesioned OA human osteochondral explants

ObjectiveTo explore IL11 co-expression profiles in our previously reported RNA-sequencing dataset of OA articular cartilage, in interaction with IL6, and to investigate the effects of hrIL11 administration as potential therapeutic strategy for OA articular cartilage using our biomimetic aged human osteochondral explant model of OA.MethodsWe used RNA-sequencing datasets of macroscopically preserved and lesioned OA articular cartilage (N = 35 patients). Spearman correlations were calculated between IL11 and IL6 expression levels and genes expressed in cartilage (N = 20048 genes). Osteochondral explants were isolated from macroscopically preserved and lesioned areas of the joint and were kept in culture for two weeks, with or without exposure to 200ng/ml hrIL11.ResultsWe found no overlap in correlating genes between IL11 and IL6, indicating their distinct roles in articular cartilage. Moreover, we identified more genes being correlated to IL11 in the lesioned compared to preserved articular cartilage (N = 203 and 106 genes, respectively). Upon treatment of ex vivo OA articular cartilage with hrIL11, we overall observed unbeneficial effects on chondrocyte phenotype, as illustrated by upregulation of MMP13, EPAS1, RUNX2, and POSTN. We did not observe significant differences in Mankin scores upon addition of hrIL11.ConclusionThe current study showed that treatment of OA articular cartilage with hrIL11 is unlikely to be beneficial despite previous indications of hrIL11 as potential druggable target. These findings underscore the importance of functionally investigating OA risk genes. Better understanding of IL11 signaling and the underlying pathways is necessary towards the development of OA treatment strategy.

PARP inhibitors differentially regulate immune responses in distinct genetic backgrounds of high-grade serous tubo-ovarian carcinoma models.

Immune checkpoint inhibitors (ICIs) have revolutionized treatment for several tumor indications without demonstrated benefit for ovarian cancer patients. To improve the therapeutic ratio of ICIs in ovarian cancer patients, several different clinical trials are testing combinations with poly (ADP-ribose) polymerase (PARP) inhibitors. Comparing the immunomodulatory effects of clinically advanced PARP inhibitors may help to identify the best partner to combine with ICIs. We examined the treatment effect of talazoparib (a PARP trapper) and veliparib (a solely PARP enzymatic inhibitor) in homologous recombination deficient (HRD) and homologous recombination proficient (HRP) high-grade serous tubo-ovarian carcinoma (HGSC) cell lines on immune-related gene expression. We discovered and validated that CXCL8, IL-6, and TNF gene expression were upregulated after talazoparib treatment in both OVCAR3 (HRD) and CAOV3 (HRP) HGSC cell lines. In contrast, veliparib treatment slightly elevated similar genes exclusively in a HRD HGSC cell line model. We expanded these studies to include olaparib, a PARP trapper less potent than talazoparib, and found effects specific to COV361 (BRCA1 mutant) and OVCAR8 (BRCA1 methylated) HGSC cells but not all HRD HGSC cell lines. Our studies also identified differences among PARP trappers versus veliparib on augmenting CXCL10 expression. Finally, we show that talazoparib modulates the CXCL10 response in cGAS-defective cell lines, independent of the cGAS-STING pathway. These mechanistic studies advance our understanding of how different PARP inhibitors affect the immune system in various genetic backgrounds.

Skin Barrier in Normal and Allergic Horses: What Do We Know?

Information on skin barrier in horses is limited. A study on the epidermal ultrastructure of normal and allergic horses documented disorganized amorphous intercellular lipids in the stratum corneum of allergic samples. These findings are similar to atopic canine and human skin. Currently, there is no published study comparing skin barrier function parameters between normal and allergic horses; thus, the functional implications of the ultrastructural changes are unknown. In normal horses, body location, gender, breed, and ambient conditions affect skin barrier parameters, such as Transepidermal Water Loss. Skin microbiome studies on normal horses have highlighted the importance of season and environmental conditions, since horses housed together share similar microbiomes. Skin dysbiosis and predominance of staphylococcus have been described in horses with pastern dermatitis. Transcriptomic studies of the epidermis of normal and allergic horses have found that lesional allergic skin has substantial transcriptomic differences when compared with healthy skin, namely downregulation of genes of tight junctions, keratins, and upregulation of serine proteases and IL-13. Keratinocytes harvested from horses with insect bite hypersensitivity show upregulation of IL-31 gene expression under stimulation. While more research is clearly needed, preliminary results seem to support skin barrier differences between normal and allergic horses.

Inhibition of BRD4 attenuated IFNγ-induced apoptosis in colorectal cancer organoids

BackgroundThis study aimed to analyze the functional role of Brd4 in colorectal cancer (CRC) organoids. Brd4 was identified as a CRC-related gene by our previous Sleeping Beauty mutagenesis transposon screening in mice. Brd4 is a transcriptional regulator that recognizes acetylated histones and is known to be involved in inflammatory responses. The role of Brd4 in CRC development remains largely unknown.MethodsWe knocked out Brd4 in tumor organoids carrying mutations in Apc and Kras to generate Brd4KO organoids, and performed RNA-seq. The response of Brd4KO organoids to IFNγ was analyzed via a cell viability assay, an apoptosis assay, and RNAseq. The results were validated by pharmacological inhibition experiments with JQ1 in human CRC organoids.ResultsIn Brd4KO organoids, the IFNγ signaling genes Il33 and Myc target genes were downregulated. The addition of IFNγ to the colon organoids induced apoptosis, but IFNγ-induced apoptosis was attenuated in the Brd4KO organoids compared with the control organoids (two-sided t-test, P < 0.05). Similar results were obtained from pharmacological inhibition with JQ1 in human CRC organoids; IL33 expression was decreased, and IFNγ-induced apoptosis was attenuated in the presence of JQ1.ConclusionsOur results showed that the inhibition of Brd4 suppressed IFNγ-induced cytotoxicity by modulating the Jak-Stat pathway. These data suggested that the inhibition of Brd4 could increase cell viability in the cancer microenvironment where IFNγ is abundant, revealing a new aspect of the molecular mechanism of CRC development. Our results may help in evaluating the application of Bet inhibitors in treating CRC. Additionally, our RNA-seq data sets will be helpful for clarifying the relationship between Brd4 and immunomodulators, such as Il33, or for studying the responses of colonic epithelial cells to IFNγ.

Associations between IL-33 gene and IL1RL1 gene variants in periodontitis

Background: Periodontitis is initiated by a dysbiosis in the subgingival microbial biofilm and can be related to host genetic factors. This study investigated association between periodontitis and single nucleotide variants (SNVs) in the IL-33 and IL1RL1 genes. Methods: This cross-sectional study involved 359 individuals from a public health service in Brazil. Structured questionnaire was used to collect health status and socioeconomic, demographic and behavioral characteristics. Periodontitis was diagnosed by clinical periodontal examination. Subgingival biofilm was collected at the deepest site of each sextant, and biofilm bacterial DNA was amplified by real time polymerase chain reaction (qPCR) to determine relative quantification of pathogens. Peripheral blood was collected for genomic DNA extraction and SNV genotyping was performed by qPCR. Logistic regression model was used to obtain association measures (95% confidence interval), by ussing the additive model. Results: The C allele variant of IL33 (rs2381416) was inversely associated with periodontitis, even after adjusting for the confounding covariates (p &lt; 0.01 ORadjusted: 0.45; CI: 0.24-0.84), and with the presence of the Aggregatibacter actinomycetemcomitans (Aa) pathogen, adjusting for the same covariates (p &lt; 0.01; ORadjusted: 0.46; CI: 0.27-0.76). Inverse association between this SNV and periodontitis was observed (p = 0.02; ORadjusted: 0.46; CI: 0.28-0.76) using additive genotypic model. Conclusions: Frequency of C allele variant of IL-33 (rs2381416) was lower in individuals with periodontitis and in individuals with relatively higher levels of Aa. Investigations of this variant as a potential predictor of the protective phenotype in the context of periodontitis are needed. This study will contribute to the training of health professionals involved in the treatment of periodontitis.

DOP072 The IL1-dependent transcription factor MEOX1 as a driver of differentiation programs in ACKR1+ endothelial cells of inflammatory bowel disease granulation tissue

Abstract Background Activated endothelial cells (ECs) are expanded in IBD tissue and express ACKR11, a chemokine receptor known to mark venular endothelium and enable immune cell trafficking. ACKR1+ ECs have been reported in diseases associated with inflammation and/or fibrosis2, although their role in stromal differentiation is unclear. We investigated the relationship between IL1 and inflammation in IBD and identified an IL1-dependent transcription factor (TF), MEOX1, known to regulate fibrosis in cardiac ischemia3 that is associated with IBD ulcers and expressed in ACKR1+ ECs. Methods We assessed IL-1 bioactivity and performed matched RNA-seq on a cohort of Crohn’s disease (n=23), ulcerative colitis (n=18) and non-IBD (n=17) patients. We performed scRNA-seq of n=208 very early onset (VEO)IBD and control intestinal patient samples. RNA-scope was performed in IBD tissue with/without ulcers. Primary colonic ECs and human umbilical vein (HUV)ECs were stimulated with IL1B +/- TGFB +/- JQ1 (a BRD4 inhibitor that knocked down IL1/MEOX1 driven cardiac fibrosis) followed by qPCR, bulk/scRNA-seq. Results In the paired IL1 proteomic and RNA-seq analyses of IBD tissue, we identified an IL1 and ulcer-specific gene signature which included the TF MEOX1. In scRNA seq analysis, MEOX1 was expressed exclusively in ECs (most robustly in ACKR1+ ECs, which co-expressed the IL1 receptor IL1R1) (Fig 1A). ACKR1+MEOX1+IL1R1+ ECs were significantly expanded in inflamed VEOIBD, especially in those with deep ulcers and IL10R deficiency, and exhibited hallmarks of cell cycle arrest, e.g. expression of CDKN1A (Fig 1B and data not shown). RNA-scope revealed abundant ACKR1+MEOX1+IL1R1+ EC structures specifically in granulation tissue (Fig 1C). In primary colonic ECs, IL1B stimulation induced CDKN1A, unlike TGFB which induced a proliferation/apoptosis phenotype (Fig 1D). In HUVECs, IL1B induced a spindle-shape/fibroblastic EC morphology and ACKR1/MEOX1 expression (data not shown). scRNA-seq revealed an IL1-induced cluster defined by ACKR1, CDKN1A and chemotactic/collagen-producing gene programs, eliminated upon addition of JQ1 (Fig 1E; data not shown). Conclusion ACKR1+ ECs define IBD granulation tissue and co-express IL1R1 as well as the TF MEOX1, which may prove to be a driver of differentiation programs downstream of IL1 signaling at the EC niche that may lead to fibrosis. In silico and in vitro data suggest IL1-driven EC programs are characterized by cell cycle arrest signatures. We hypothesize that the IL1/MEOX1 pathway in ACKR1+ ECs may represent a novel molecular mechanism related to ulcer biology and wound repair in IBD with potential therapeutic implications.

P0172 In silico drug repurposing approach for the discovery of therapeutics for intestinal fibrosis in crohn’s disease

Abstract Background Intestinal fibrosis in Crohn’s disease (CD) can lead to stricture formation and other disease-related complications.1,2 Given the lack of anti-fibrotic drugs, current treatment strategies are limited to anti-inflammatory therapies, endoscopic balloon dilation and surgery.1 Our aim was to identify compounds and druggable targets that can reverse the gene expression signature in mucosal fibroblasts associated with intestinal fibrosis. Methods A signature associated with fibrotic CD was derived from three publicly available transcriptomic datasets of fibroblasts isolated from fibrostenotic strictures of CD patients. We conducted a meta-regression analysis across the transcriptomic datasets to identify significant differentially expressed genes (DEGs) associated with fibrostenotic CD. Three drug repurposing platforms (iLINCS, L1000 CDS2 and CLUE io), connected to the NIH LINCS database,3,4 were used to identify compounds that could reverse the fibrotic signature, ranked by their anti-fibrotic potential using the CoDReS (Composite Drug Reranking Scoring) tool. Transcription factors and miRNAs targeting the fibrostenotic signature were identified via the TRRUST and miRWalk databases. A gene interaction network was constructed, incorporating transcription factors (TRRUST TF), miRNAs and DEGs. Drugs targeting key network components were identified using the DGiDb database. Results Via the analysis of combined transcriptomic datasets from fibrostenotic- versus non-stenosis-associated mucosal fibroblasts, fibroblast activation protein (FAP), IL7 receptor and transcription factor AP-2 gamma, emerged as the most significantly upregulated genes in fibrostenotic CD (Figure 1). Out of 6,783 compounds, PI3K/Akt/mTOR inhibitors, Src kinase family inhibitors and histone deacetylase inhibitors were predicted as being most effective in reversing this pathologic gene signature. The gene interaction network revealed 15 hub genes as central components with the most interconnections (Table 1). Among these top 15 key gene regulators, IL6, PPARγ and angiotensin receptor type 1 (AGTR1) were the highest ranked druggable targets, based on the drug-gene interaction analysis. Conclusion Our in silico drug repurposing approach identified several potential anti-fibrotic compounds and druggable targets for the treatment of CD-associated intestinal fibrosis. These findings open novel avenues for the development of anti-fibrotic drugs and provide a foundation for future research on cell and pathway-specific mechanisms driving fibrosis in CD.