IL23R Gene Research Articles

Lack of association between polymorphisms in interleukin (IL)-12, IL-12R, IL-23, IL-23R genes and Takayasu arteritis in a Chinese population.

The goal of this study was to investigate the relationship between polymorphisms in interleukin (IL)-12, IL-12R, IL-23, and IL-23R genes and Takayasu arteritis (TA) in a Chinese population. A case-control study was performed to investigate the associations of 19 single nucleotide polymorphisms (SNPs) mapping to IL12A, IL12B, IL12RB1, IL12RB2 and IL23R with susceptibility to TA in 145 Chinese TA patients and 300 healthy controls. Genotype identification was performed with the MassARRAY system from Sequenom. The statistical analysis was conducted by Chi square test and unconditional logistic regression with plink. No significant differences were found for the distribution of allele and genotype frequencies of these SNPs between TA patients and healthy controls. However, a trend for IL12A rs582054 and IL23R rs1004819 in association with the TA phenotype was detected. TA patients carrying the rs582054/rs568408 haplotype (P'=0.019) appeared less likely to progress to a more severe form of disease. And the C allele (P'=0.082) of IL23R rs1004819 appeared to be a protective factor to refractory disease. These findings suggest that the polymorphisms of IL12A, IL12B, IL12RB1, IL12RB2 and IL23R might make no contribution to the susceptibility of TA in the Chinese population.

Generation and Characterization of Mice Expressing a Conditional Allele of the Interleukin-1 Receptor Type 1

The cytokines IL-1α and IL-1β exert powerful pro-inflammatory actions throughout the body, mediated primarily by the intracellular signaling capacity of the interleukin-1 receptor (IL-1R1). Although Il1r1 knockout mice have been informative with respect to a requirement for IL-1R1 signaling in inflammatory events, the constitutive nature of gene elimination has limited their utility in the assessment of temporal and spatial patterns of cytokine action. To pursue such questions, we have generated C57Bl/6J mice containing a floxed Il1r1 gene (Il1r1loxP/loxP), with loxP sites positioned to flank exons 3 and 4 and thereby the ability to spatially and temporally eliminate Il1r1 expression and signaling. We found that Il1r1loxP/loxP mice breed normally and exhibit no gross physical or behavioral phenotypes. Moreover, Il1r1loxP/loxP mice exhibit normal IL-1R1 receptor expression in brain and spleen, as well as normal IL-1R1-dependent increases in serum IL-6 following IL-1α injections. Breeding of Il1r1loxP/loxP mice to animals expressing a cytomegalovirus (CMV)-driven Cre recombinase afforded efficient excision at the Il1r1 locus. The Il1r1loxP/loxP line should be a valuable tool for the assessment of contributions made by IL-1R1 signaling in diverse cell types across development.

Ancestry informative markers and selected single nucleotide polymorphisms in immunoregulatory genes on preterm labor and preterm premature rupture of membranes: a case control study.

BackgroundA genetic predisposition to Preterm Labor (PTL) and Preterm Premature Rupture of Membranes (PPROM) has been suggested; however the relevance of polymorphisms and ancestry to susceptibility to PTL and PPROM in different populations remains unclear. The aim of this study was to evaluate the contribution of maternal and fetal SNPs in the IL1B, IL6, IL6R, TNFA, TNFR, IL10, TLR2, TLR4, MMP9, TIMP1 and TIMP2 genes and the influence of ancestry background in the susceptibility to PTL or PPROM in Brazilian women.MethodsCase–control study conducted at a tertiary hospital in São Paulo State, Brazil. We included women with PTL or PPROM and their babies (PTL: 136 women and 88 babies; PPROM: 65 women and 44 babies). Control group included 402 mother-babies pairs of term deliveries. Oral swabs were collected for identification of AIMs by fragment analysis and SNPs by Taqman® SNP Genotyping Assays and PCR. Linkage Disequilibrium and Hardy-Weinberg proportions were evaluated using Genepop 3.4. Haplotypes were inferred using the PHASE algorithm. Allele, genotype and haplotype frequencies were compared by Fisher’s exact test or χ2 and Odds Ratio. Logistic regression was performed. Clinical and sociodemographic data were analyzed by Fisher’s exact test and Mann–Whitney.ResultsPTL was associated with European ancestry and smoking while African ancestry was protective. The fetal alleles IL10-592C (rs800872) and IL10-819C (rs1800871) were also associated with PTL and the maternal haplotype TNFA-308G-238A was protective. Maternal presence of IL10-1082G (rs1800896) and TLR2A (rs4696480) alleles increased the risk for PPROM while TNFA-238A (rs361525) was protective. Family history of PTL/PPROM was higher in cases, and time to delivery was influenced by IL1B-31T (rs1143627) and TLR4-299G (rs4986790).ConclusionThere is an association between European ancestry and smoking and PTL in our Brazilian population sample. The presence of maternal or fetal alleles that modify the inflammatory response increase the susceptibility to PTL and PPROM. The family history of PTL/PPROM reinforces a role for genetic polymorphisms in susceptibility to these outcomes.

INVESTIGATION OF CANDIDATE GENE POLYMORPHISMS IN AN IMMUNE RESPONSE AS MARKERS FOR THE RISK OF DEVELOPING RHEUMATOID ARTHRITIS AND PRODUCING AUTOANTIBODIES

Objective: to investigate the distribution of the genotypes and alleles of the PTPN22, TNFAIP3, CTLA4, TNFA, IL6, IL6R, IL10, MCP1, and ICAM1 genes in patients with rheumatoid arthritis (RA) and in the control group of healthy individuals, to estimate their significance as molecular genetic markers for predisposition to RA; and to analyze the correlation between the gene polymorphisms included in the study and the production of anti-cyclic citrullinated peptide antibodies (ACCPA) and IgM rheumatoid factor (RF). Subjects and methods. The investigation was conducted within the framework of the «Early arthritis: Diagnosis, outcome, criteria, active treatment program». The prospective follow-up study included 122 patients with RA fulfilling the 1987 American College of Rheumatology (ACR) criteria; with disease duration of ≤ 2 years. 73 (59.8%) patients were included during the first 6 months after the onset of the disease. 74 (60.7%) and 81 (66.5%) patients were found to be positive for ACCPA and IgM RF, respectively. 314 healthy blood donors served as a control group. A real-time polymerase chain reaction was used in the patients and control individuals to study the distribution of the polymorphic variants of PTPN22 (+1858 C >T, rs2476601), TNFAIP3 (rs675520, rs6920220, rs10499194), CTLA4 (+49A>G, rs231775 ), TNFА (-308A>G, rs1800629), IL6 (-174G>C, rs1800795), IL6R (+358A>C, rs8192284), IL10 (-592A>C, rs1800872, -1082 A>G, rs1800896), MCP1/CCL2 (+2518A>G, rs1024611), and ICAM1 (721G>A, rs1799969) genes. Results and discussion. This analysis revealed an association of PTPN22 (+1858 C >T, rs2476601) and TNFAIP3 (rs675520, rs10499194) polymorphisms with the risk of RA (odds ratio (OR), 1.5; 95% confidence interval (CI), 1.0–2.3; p = 0.05; OR, 1.5; 95% CI, 1.1–2.0; p = 0.02; OR, 0.5; 95% CI, 0.4–0.8; p = 0.01, respectively. Further, there was a tendency towards a positive association of TNFAIP3 (rs6920220) and IL6R (rs8192284) polymorphisms with a predisposition to RA (p = 0.056). IL6 (rs1800795), IL10 (rs1800872, rs1800896), MCP1/CCL2 (rs1024611), and ICAM1 (rs1799969) polymorphisms were not associated with the risk of RA. An analysis of the findings after patient stratification by ACCPA and IgM RF (a binary variable) showed that none of the polymorphisms in question was associated with RF state. At the same time, PTPN22 (rs2476601), TNFAIP3 (rs675520), TNFAIP3 (rs10499194), and TNFА (rs1800629) polymorphisms were found to be significantly related to ACCPA state (a binary variable). The level of ACCPA as a quantitative variable was statistically significantly associated with CTLA4 (rs231775) and TNFА (rs1800629) polymorphisms in a dose-dependent fashion (р = 0.025 and р = 0.015, respectively). There was a marked tendency towards an association of ACCPA levels and IL6R gene polymorphism (p = 0.07). IL6 (rs1800795), IL10 (rs1800872, rs1800896), MCP1/CCL2 (rs1024611), and ICAM1 (rs1799969) polymorphisms were not correlated with ACCPA state (binary and quantitative variables). Conclusion. The findings suggest that a number of genes are implicated in the pathogenesis of RA and that they are involved in the development of ACCPA-positive and ACCPA-negative RA subtypes. No relationship was found between the production of IgM RF and the polymorphisms of the genes under study. The findings suggest that there appears to be different mechanisms for the formation of autoantibodies (ACCPA and IgM RF) in RA.

Assessment of the Effects of IL9, IL9R, IL17A, and IL17F Gene Polymorphisms on Women with Allergic Rhinitis in Shahrekord, Iran.

Background:The genes encoding IL9, IL9R, IL17A, and IL17F have recently been implicated in the genetic basis of rhinitis and allergy.Aim:The purpose of this study was to assess the association of the single nucleotide polymorphisms (SNPs) of IL9, IL9R, IL17A, and IL17F and potential interaction of these genes with the determination of IgE levels in women with allergic rhinitis (AR) in Shahrekord, Iran.Subjects and Methods:In a case–control study, SNPs from the IL9, IL9R, IL17A, and IL17F were genotyped in 394 random samples including 195 AR patients and 199 normal controls. Enzyme-linked immunosorbent assay was performed for the determination of serum total IgE levels. The Student's t-test was used to compare the differences. The Chi-square test was performed to compare proportions of cases with different clinical features among cases with different genotypes. The genotype and allele frequencies were obtained by direct counting. Hardy–Weinberg equilibrium was tested between cases and controls separately. The relative risk associated with rare alleles was estimated as an odds ratio with 95% confidence interval. P ≤ 0.05 was considered statistically significant.Results:The rs731476 SNP in the IL9R was significantly associated with the AR phenotype in women. No association was found between any of the other SNPs in IL9, IL17A, and IL17F genes and AR. In the gene–gene interaction analysis, we found that IL9R/IL9 genotype rs731476 T-/rs2069885 G conferred a higher risk for AR phenotype development. We also did not find a significant association in terms of IgE levels between cases and controls.Conclusion:Our result suggests that the rs731476 SNP located in the IL9R is associated with an increased susceptibility to AR in females. In a subsequent gene–gene interaction analysis, the rs731476 T-/rs2069885 G-genotype combination (IL9R/IL9) has significantly been associated with the development of the AR phenotype.

Transcriptome Analysis of Erythroid Cells Cultured from Diamond Blackfan Anemia Patients with Ribosomal and GATA1 Mutations Reveals Dysregulation of Inflammatory Response Genes

Diamond Blackfan anemia (DBA) is a rare, congenital bone marrow failure syndrome characterized by red cell aplasia, usually without perturbation of other hematopoietic lineages. DBA patients are generally diagnosed during infancy or early childhood, have a high frequency of congenital anomalies, and a predisposition to cancer. Approximately 65% of DBA patients have identifiable heterozygous gene mutations or deletions in ribosomal protein genes. Additionally, mutations in GATA1, a key transcription factor in erythropoiesis, have been demonstrated in DBA patients (Sankaran VG et al. JCI. 122: 2439-43, 2012; Ludwig LS et al. Nat Med 20(7):748-53, 2014; Klar J et al. Br J Haem 166(6): 949-51, 2014). Despite our knowledge of the molecular pathology, the mechanism underlying the erythroid failure in DBA is not well understood, largely due to the inherent difficulties in studying primary erythroid cells from DBA patients. Furthermore, DBA patients with GATA1 mutations have not been well characterized and it remains unclear whether the pathogenic mechanisms of GATA1 DBA are similar to that of ribosomal DBA.We designed a two-step, 14 day in vitro culture system to generate erythroid cells from CD34+ stem/progenitor cells isolated from <10ml of peripheral blood (PB) collected from patients enrolled in the DBA Registry of North America. Patients with RPL5, RPS17(n=2), RPS24 and RPL35a, GATA1 mutations (n=2), and patients without identifiable mutations (n=3) were studied and compared to healthy control PB CD34+ cells. At day 14, we routinely obtain at least 10x fewer CD235+ erythroid cells (proerythroblasts and basophilic erythroblasts) in DBA cultures vs. controls (1x106 vs. 1x107 from 1x104 CD34+ cells). Further, a 2-7 day delay in the acquisition of CD235 was observed.Gene expression analyses was performed by Affymetrix GeneChip Human Gene ST Arrays and RNASeq to analyze protein coding and long non-coding RNA transcripts in cells from day 14 of erythroid cultures. Ingenuity pathway analysis (IPA) of the dysregulated genes between patients with ribosomal mutations, GATA1 mutations and controls revealed that many of the genes dysregulated in DBA were involved in multiple leukocyte migration and inflammatory signaling pathways, including the IL8, IL1R1, CXCR4, ICAM3, MPO, TNFSF10, and TLR4 genes with IL6, TNF, and lipopolysaccharide as top upstream regulators. Notably, the dysregulated genes in GATA1 patient cells largely overlapped that of the DBA patients with ribosomal mutations, including disruption of the leukocyte migration and inflammatory response genes. Patients with GATA1 and ribosomal protein mutations shared a number of dysregulated erythroid genes including AHSP, FAM132B, HEMGN, and TRIM10, however GATA1 patient cells showed GATA1 as the top upstream regulator and additionally showed dysregulation of heme biosynthesis pathway genes, including the ALAS2, FECH, CPOX, PPOX, and UROS.Few studies have looked at the pathogenesis of DBA in patients with GATA1 mutations. The DBA-associated splice donor mutation in GATA1 results in the exclusive expression of the short form of the GATA1 protein (GATA1s), which lacks the transactivation domain. Western blot analysis of control erythroid cells showed expression of both full length GATA1 and GATA1s, with the full length protein predominating. In cells from the patients with the GATA1 mutation, only GATA1s protein was expressed and the level of GATA1s exceeded the combined level of both GATA1 isoforms in control cells. Erythroid cells from a patient with an RPL5 mutation showed GATA1 protein levels comparable to controls, indicating the DBA phenotype of the RPL5 patient is not due to altered translation of full length GATA1. Northern blot analysis demonstrated that the GATA1 mutation did not affect ribosomal RNA processing.In summary, our transcriptome analyses have revealed novel insights into the molecular pathogenesis of ribosomal and GATA1-mutated DBA. Significant dysregulation of inflammatory gene pathways in DBA patients with both ribosomal protein and GATA1 mutations were observed, suggesting a shared pathological mechanism. Furthermore, the heme biosynthesis pathway was uniquely disrupted in patients with GATA1 mutations. Further investigating the inflammatory pathways in DBA may reveal novel targets for therapeutic development. DisclosuresNo relevant conflicts of interest to declare.

Immunophenotyping with CD135 and CD117 predicts the FLT3, IL-7R and TLX3 gene mutations in childhood T-cell acute leukemia

With the combination of immunophenotyping and molecular tests, it is still a challenge to identify the characteristics of T cell acute lymphoblastic leukemia (T-ALL) associated with distinct outcomes. This study tests the possible correlation of cellular expression of CD135 and CD117 with somatic gene mutations in T-ALL. One hundred sixty-two samples were tested, including 143 at diagnosis, 15 from T-lymphoblastic lymphoma at relapse, and four relapse samples from sequential follow-up of T-ALL. CD135 and CD117 monoclonal antibodies were included in the T-ALL panel of flow cytometry. The percentage of cells positivity and the median fluorescence intensity were correlated with gene mutational status. STIL-TAL1, TLX3, FLT3 and IL7R mutations were tested using standard techniques.STIL-TAL1 was found in 24.8%, TLX3 in 12%, IL7R in 10% and FLT3-ITD in 5% of cases. FLT3 and IL7R mutations were mutually exclusive, as were FLT3-ITD and STIL-TAL1. Associations of CD135high (p<0.01), CD117intermediate/high (p=0.02) and FLT3-ITD, CD117low with IL7Rmutated (p<0.01) and CD135high with TLX3pos were observed.We conclude that the addition of CD135 and CD117 to the diagnosis can predict molecular aberrations in T-ALL settings, mainly segregating patients with FLT3-ITD, who would benefit from treatment with inhibitors of tyrosine.

Epistasis analysis links immune cascades and cerebral amyloidosis.

BackgroundSeveral lines of evidence suggest the involvement of neuroinflammatory changes in Alzheimer’s disease (AD) pathophysiology such as amyloidosis and neurodegeneration. In fact, genome-wide association studies (GWAS) have shown a link between genes involved in neuroinflammation and AD. In order to further investigate whether interactions between candidate genetic variances coding for neuroinflammatory molecules are associated with brain amyloid β (Aβ) fibrillary accumulation, we conducted an epistasis analysis on a pool of genes associated with molecular mediators of inflammation.Methods[18F]Florbetapir positron emission tomography (PET) imaging was employed to assess brain Aβ levels in 417 participants from ADNI-GO/2 and posteriorly 174 from ADNI-1. IL-1β, IL4, IL6, IL6r, IL10, IL12, IL18, C5, and C9 genes were chosen based on previous studies conducted in AD patients. Using the [18F]florbetapir standardized uptake value ratio (SUVR) as a quantitative measure of fibrillary Aβ, epistasis analyses were performed between two sets of markers of immune-related genes using gender, diagnosis, and apolipoprotein E (APOE) as covariates. Voxel-based analyses were also conducted. The results were corrected for multiple comparison tests. Cerebrospinal fluid (CSF) Aβ1-42/phosphorylated tau (p-tau) ratio concentrations were used to confirm such associations.ResultsEpistasis analysis unveiled two significant single nucleotide polymorphism (SNP)-SNP interactions (false discovery rate (FDR) threshold 0.1), both interactions between C9 gene (rs261752) and IL6r gene (rs4240872, rs7514452). In a combined sample, the interactions were confirmed (p ≤ 10–5) and associated with amyloid accumulation within cognitively normal and AD spectrum groups. Voxel-based analysis corroborated initial findings. CSF biomarker (Aβ1-42/p-tau) confirmed the genetic interaction. Additionally, rs4240872 and rs7514452 SNPs were shown to be associated with CSF and plasma concentrations of IL6r protein.ConclusionsCertain allele combinations involving IL6r and C9 genes are associated with Aβ burden in the brain. Hypothesis-driven search for epistasis is a valuable strategy for investigating imaging endophenotypes in complex neurodegenerative diseases.Electronic supplementary materialThe online version of this article (doi:10.1186/s12974-015-0436-z) contains supplementary material, which is available to authorized users.

Atrial Fibrillation in Patients With Chronic Obstructive Pulmonary Disease Is Associated With Polymorphism of Interleukin 6 Gene

It can be suggested that development of atrial fibrillation (AF) in patients with chronic obstructive pulmonary disease (COPD) is directly related to the system of inflammation. Genetic polymorphism of factors of this system can be one of components of mechanism of AF in COPD. Aim: to elucidate polymorphic markers of genes of factors of the system of inflammation associated with AF in patients with COPD. Material and methods. We examined 208 patients with COPD (52 with and 156 without AF). Examination included spirometry, echocardiography, and study of frequencies of polymorphic markers G(-238)A, G(-308)A of tumor necrosis factor (TNF) gene, C(-819)T of interleukin (IL) 10 gene, G(-174)C of IL-6 gene, rs2228145(AC) of IL-6R gene, and rs2069762(A/C) of IL-2 gene. RESULTS. Factors associated with AF were left atrial volume (odds ratio [OR] 1.021, 95% confidence interval [ClI] 1.004-1.043, p = 0,027), right atrial volume (OR 1.02, 95% CI 1.001-1.040, p=0.021), and carriage of C allele of polymorphic marker G(-174)C of IL-6 gene (OR 6.02, 95% Cl 1.87-19.38, p = 0.003). C allele of polymorphic marker G(-174)C of IL-6 gene can be considered to be independently associated with development of AF in patients with COPD.

Association study of functional polymorphisms in interleukins and interleukin receptors genes: IL1A, IL1B, IL1RN, IL6, IL6R, IL10, IL10RA and TGFB1 in schizophrenia in Polish population

Schizophrenia has been associated with a large range of autoimmune diseases, with a history of any autoimmune disease being associated with a 45% increase in risk for the illness. The inflammatory system may trigger or modulate the course of schizophrenia through complex mechanisms influencing neurodevelopment, neuroplasticity and neurotransmission. In particular, increases or imbalance in cytokine before birth or during the early stages of life may affect neurodevelopment and produce vulnerability to the disease. A total of 27 polymorphisms of IL1N gene: rs1800587, rs17561; IL1B gene: rs1143634, rs1143643, rs16944, rs4848306, rs1143623, rs1143633, rs1143627; IL1RN gene: rs419598, rs315952, rs9005, rs4251961; IL6 gene: rs1800795, rs1800797; IL6R gene: rs4537545, rs4845617, rs2228145, IL10 gene: rs1800896, rs1800871, rs1800872, rs1800890, rs6676671; IL10RA gene: rs2229113, rs3135932; TGF1B gene: rs1800469, rs1800470; each selected on the basis of molecular evidence for functionality, were investigated in this study. Analysis was performed on a group of 621 patients with diagnosis of schizophrenia and 531 healthy controls in Polish population. An association of rs4848306 in IL1B gene, rs4251961 in IL1RN gene, rs2228145 and rs4537545 in IL6R with schizophrenia have been observed. rs6676671 in IL10 was associated with early age of onset. Strong linkage disequilibrium was observed between analyzed polymorphisms in each gene, except of IL10RA. We observed that haplotypes composed of rs4537545 and rs2228145 in IL6R gene were associated with schizophrenia. Analyses with family history of schizophrenia, other psychiatric disorders and alcohol abuse/dependence did not show any positive findings. Further studies on larger groups along with correlation with circulating protein levels are needed.

Investigation of association of the IL-12B and IL-23R genetic variations with psoriatic risk in a South Indian Tamil cohort

BackgroundPsoriasis is a T-cell mediated chronic systemic inflammatory skin disease. Emerging evidences suggest the interleukin (IL)-12B and IL-23R genes encoding the common p40 subunit of IL-12 and IL-23 are the key cytokines in T-helper (Th)1 and Th17 differentiation and function. Certain allelic variants of these genes significantly influence the risk of psoriasis. Hence we undertook to study the association of IL-12B and IL-23R gene polymorphisms with disease susceptibility in South Indian Tamil patients with psoriasis. Methods360 psoriatics and 360 healthy controls were included in this case control study. IL-12B gene (rs3212227) and IL-23R gene (rs2201841, rs10889677 and rs11805303) polymorphisms were typed by using TaqMan 5′allele discrimination assay and cytokine levels were assayed by ELISA. ResultsWe observed that the patients carrying the risk genotypes of IL-12B (rs3212227) and IL-23R (rs2201841) conferred an increased susceptibility to psoriasis. We did not find any significant association between IL-23R (rs10889677 and rs11805303) gene polymorphisms and psoriasis risk in South Indian Tamil population. We did not observe any significant difference in haplotypes between the psoriasis cases and controls. We observed a significant increase in the mean IL-23 levels in psoriatics and the higher levels of IL-23 were found in the minor variant genotype CC when compared with that of heterozygous CT and major variant TT genotypes of rs2201841. Individual genotypes of rs10889677 and rs11805303 and IL-12 (rs3212227) were not significantly associated with their plasma levels. ConclusionOur results suggest that IL-12B (rs3212227) and IL-23R (rs2201841) polymorphisms confer increased risk of psoriasis in our ethnic South Indian Tamils.

DNA methylation signature of interleukin 1 receptor type II in asthma.

Interleukin 1 and its receptors are associated with allergic diseases such as asthma. In the present study, we measured DNA methylation at the IL1R1 and IL1R2 gene loci and assessed for associations with asthma-related phenotypes and gene expressions. We found that asthmatic and atopic individuals have higher IL1R2 promoter DNA methylation than control subjects. Additionally, we observed a negative correlation between DNA methylation at the IL1R2 promoter and IL1R2 mRNA expression. These results suggest for the first time that IL1R2 promoter DNA methylation is associated with its gene repression in allergic diseases such as asthma.Electronic supplementary materialThe online version of this article (doi:10.1186/s13148-015-0114-0) contains supplementary material, which is available to authorized users.

Abstract 3530: Gene expression profile of OTX015, a BET bromodomain inhibitor, in preclinical models of non-small-cell lung cancer (NSCLC) and small-cell lung cancer (SCLC) models

Abstract Background. BET bromodomain proteins recognize chromatin modifications and act as epigenetic readers contributing to gene transcription. OTX015, a novel BET bromodomain inhibitor currently in clinical Phase Ib studies in hematologic malignancies and solid tumors, including lung cancer. We have shown in vitro and in vivo activity of OTX015 in NSCLC and SCLC models (Riveiro et al; EORTC 2014), however the mechanism of action and relevant affected genes are not fully characterized. This study aimed to elucidate pathways and genes affecting response/resistance to OTX015 in lung models.Methods. OTX015 (OncoEthix SA, Switzerland), GI50 values were determined with the MTT assay after 72h exposure in 5 NSCLC cell lines (H2228, H3122, A549, HOP62, HOP92) and 4 SCLC cell lines (H82, H69, DMS79, DMS114). For cell cycle analysis, cells were stained with propidium iodide and analyzed with a FACScan flow cytometer. RNA was isolated after treatment with DMSO or OTX015 (500 nM) for 2, 4, 8 or 12 and 24h and gene expression profiles (GEP) were obtained in all NSCLC and 2 SCLC (H82 and DMS114) cell lines with Illumina HumanHT-12 Expression BeadChips. Results. OTX015 displayed antiproliferative effects in 4 of the 5 NSCLC cell lines with GI50 values from 110 to 940 nM, whereas A549 cells (harboring mutations in KRAS and LKB1 genes) were resistant to OTX015. Among SLCL models, DMS114 cells showed dose-dependent sensitivity to OTX015 [GI50 = 120nM], while H82, H69, and DMS79 cells were resistant [GI50 &amp;gt;6 μM], despite overexpressing CMYC and/or NMYC. In sensitive-NSCLC models we observed an increase in cells in the S phase after OTX015 treatment, whereas an increase in the G1 phase was observed in the sensitive DMS114 cell line. GEP showed that changes in expression levels of EFR3B and genes related to RNA biogenesis were higher in SCLC cells with reduced sensitivity to OTX015 than in sensitive cells. On the other hand, genes related to histones and chromatin structure were the most differentially expressed after OTX015 exposure in both sensitive and resistant cell lines. In NSCLC models, EFR3B and FOS gene up-regulation, IL7R and IL6-related gene down-regulation were associated with higher OTX015 sensitivity Expression changes more prominent in OTX015-resistant cells affected the transcription factor FOXD1 and genes related to RNA processing. Conclusions. Our data contribute to elucidate the mechanisms associated with OTX015 sensitivity in several lung cancer in vitro models, supporting the rationale for the ongoing clinical Phase Ib study in solid tumors including NSCLC patients (KRAS+; ALK+). Citation Format: Maria Eugenia Riveiro, Ivo Kwee, Lucile Astorgues-Xerri, Mohamed Bekradda, Ramiro Vazquez, Andrea Rinaldi, Esteban Cvitkovic, Francesco Bertoni. Gene expression profile of OTX015, a BET bromodomain inhibitor, in preclinical models of non-small-cell lung cancer (NSCLC) and small-cell lung cancer (SCLC) models. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3530. doi:10.1158/1538-7445.AM2015-3530

Interleukin-23 Receptor Gene Polymorphism May Enhance Expression of the IL-23 Receptor, IL-17, TNF-α and IL-6 in Behcet's Disease.

PurposeRecent studies identified an association between Behcet’s disease (BD) and the IL-23R gene polymorphism (rs17375018) in different populations. This study examined whether this IL-23R gene polymorphism is associated with enhanced inflammatory responses.MethodsWe recruited 27 BD patients and 32 controls with three genotypes. Peripheral blood mononuclear cells (PBMCs) were seeded with or without anti-CD3 and CD28. Cells were incubated for 24 hours, and then supernatants were collected and stored at −20◦C until analyzed. Levels of interferon (IFN)-γ, tissue necrosis factor (TNF)-α, interleukin (IL)-17 and IL-6 were detected by ELISA. IL-23R expression was assessed by quantitative real-time polymerase chain reaction (RT-PCR).ResultsThe expression of IL-23R was significantly higher in both BD patients and healthy controls with the GG genotype compared to the AG and AA genotype with anti-CD3 and CD28 stimulation (all P-value < 0.05). Among the PBMCs cultured with anti-CD3 and CD28 stimulation, there was an elevated secretion of TNF-α, IL-6 and IL-17 in BD patients and healthy controls with the GG genotype. However, there was no significant change in secretion of IFN- γ in BD patients and healthy controls among the genotype of this IL-23R gene polymorphism.ConclusionsThe results suggest that the GG genotype of the rs17375018 variant in the IL-23R gene enhances pro-inflammatory cytokine responses.

Involvement of IL17A, IL17F and IL23R Polymorphisms in Colorectal Cancer Therapy.

IL23/IL17 pathway plays an important role in the development of inflammatory bowel diseases (IBD). In general, the genes encoding the cytokines are genetically polymorphic and polymorphisms in genes IL23R and IL17 have been proved to be associated with its susceptibility to inflammatory diseases as well as cancer including colorectal cancer. Moreover, it has been shown that these interleukins are involved in anti-tumor or pro-tumor effects of various cancers. Previously, we showed that there is a significant association between IL17A, IL17F and IL23R polymorphisms as well as the occurrence of colorectal cancer and the clinical features of the disease. The purpose of the present work is to investigate an association between IL17A, IL17F and IL23R polymorphisms in 102 Tunisian patients with colorectal cancer treatment. The association was analyzed by statistical tools. We found that patients with mutated genotypes of IL17A G197A SNP could be a risk factor for the inefficiency of chemotherapy and radiotherapy. Unlike IL17F variant, patients with wild type genotypes require surgery and adjuvant chemotherapy. On the one hand, we found no evidence that supports a significant association between IL23R polymorphism and the combined genotypes of these three genes and the colorectal cancer treatment. On the other hand, we showed that there is an important interaction between IL17A/IL17F polymorphisms and the stage of the disease as well as its treatment. Finally, patients with IL17F wild type genotype highlighted that there is a valid longer OS without all treatments and with radiotherapy and a neoadjuvant chemotherapy. In contrast, we observed that there are no relationships between IL17A, IL23R and the survival of these patients neither with nor without the treatment. Our results suggest that polymorphisms in IL17A and IL17F genes may be a predictive source of colorectal cancer therapy type. Therefore, IL17F may serve as an independent prognostic factor for overall survival in patients with colorectal cancer.

T helper cell‑related cytokine gene polymorphisms and vitamin D pathway gene polymorphisms as predictors of survival probability in patients on renal replacement therapy.

Negative outcomes in patients on renal replacement therapy (RRT) may have a source in T helper (Th)-cell imbalance or vitamin D deficiency. We examined the association of genes encoding cytokines related to Th1 and Th2 cells and vitamin D pathway genes with survival probability of patients on RRT. The study included 1253 patients on hemodialysis. IL13, IL4R, IL18, IL12A, IL12B, IL28B, MCP1, GC, VDR, and RXRA gene polymorphisms were tested. The Kaplan-Meier method with the log‑rank test was used to estimate significance of survival probabilities. Patients carrying the IL13 minor T allele had an increased risk of death compared with CC subjects (log‑rank test, P = 0.005; hazard ratio [HR], 1.40; 95% confidence interval [CI], 1.11-1.76; P = 0.005). IL28B rs8099 917 GG patients had higher mortality rates compared with IL28B GT+TT carriers (log‑rank test, P = 0.04; HR, 1.82; 95% CI, 1.10-3.01; P =0.02). IL28B rs12979860 TT carriers had an increased risk of death compared with CC+CT carriers (log‑rank test, P = 0.02; HR, 1.62; 95% CI, 1.09-2.42; P = 0.02) only when they were negative for hepatitis B virus (HBV) or hepatitis C virus (HCV) infection. The prevalence of coronary artery disease differed significantly among patients with the IL28B rs12979860 TT genotype compared with CC+CT carriers (odds ratio, 1.87; 95% CI, 1.14-3.09; P = 0.01). There was no association between the GC, VDR, and RXRA nucleotide variants and survival. The IL13 rs20541 T allele and IL28B rs8099917 GG genotype are negative predictors of survival in patients on RRT, while the IL28B rs12979860 TT genotype increases the risk of death only in patients negative for HBV or HCV infection.

Investigation of interleukin-12, interleukin-17 and interleukin-23 receptor gene polymorphisms in alopecia areata.

To investigate the distribution of interleukin (IL)-12 (IL12; 1188A/C), IL17 (A7488G) and IL-23 receptor (IL23R; +2199A/C) gene polymorphisms in patients with alopecia areata. Patients with alopecia areata and healthy controls were enrolled in this case-control study. Genotyping of the IL12 (1188A/C), IL17 (A7488G) and IL23R (+2199A/C) polymorphisms was undertaken. Genotype frequencies were compared between the two groups. The study enrolled 100 patients with alopecia areata and 71 control subjects. No significant differences were found in the frequencies for the IL12 and IL23R gene polymorphisms between the patient and control groups. The IL17 GG genotype was significantly more common and the IL17 GA genotype was significantly less common in patients with alopecia areata compared with controls, but only 10% of patients had the GG genotype. The IL17 GG genotype was associated with susceptibility for alopecia areata, but this genotype was only present in a small number of patients. The IL12 and IL23R gene polymorphisms were not found to have a significant association with alopecia areata.

Polymorphisms of IL-23 receptor gene are associated with susceptibility to pulmonary tuberculosis and drug-resistant pulmonary tuberculosis

To explore the associations of interleukin-23 (IL-23) receptor gene polymorphisms, susceptibility to pulmonary tuberculosis and drug-resistant pulmonary tuberculosis (TB). For this paired case-control study, 13 single nucleotide polymorphisms (SNPs) in IL-23R gene were genotyped by multiplex SNaPshot with samples from 250 pairs of culture positive pulmonary TB patients and healthy controls. And the relationship between genotype, allele, haplotype and susceptibility to pulmonary tuberculosis was analyzed. Two SNPs of IL-23R gene (rs10889677 and rs7518660) (OR = 1.53, 95% CI: 1.01-2.31; OR = 6.25, 95% CI: 3.85-10.00) were significantly associated with susceptibility to pulmonary TB and one SNP (rs11465802) (OR = 2.90; 95% CI: 1.69-4.98) was associated with drug-resistant pulmonary TB in the Chinese Uygur population. Two SNPs of IL-23R gene, rs10889677 and rs7518660, may be the risk factors of pulmonary tuberculosis. And rs11465802 may be a risk factor of drug-resistant pulmonary tuberculosis in Chinese Uygur population.

Polymorphisms in the promoter of interleukin-12β2 and interleukin-23 receptor genes influence milk production traits in Chinese Holstein cows

Interleukin-12 ( IL-12 ) and interleukin-23 ( IL-23 ) are proinflammatory cytokines produced by macrophages and dendritic cells in response to infection with intracellular pathogens. Given the importance of IL-12 and IL-23 for modulating inflammation and the host immune response, the IL-12 and IL-23 receptor genes may be suitable candidate genes for studying disease resistance in dairy cattle. Twenty Chinese Holstein cows were selected randomly for PCR amplification and sequencing, and used for SNP discovery in the bovine IL-12Rβ2 and IL-23R promoter region. One SNP( c.-246G>T ) in IL-12Rβ2 gene and 2 SNPs( c.-856A>G and c.-207T>C ) in IL-23R gene were identified. Chinese Holstein cows ( n =866) were then genotyped using Sequenom MassARRAY (Sequenom Inc., San Diego, CA) based on the 3 identified SNPs, and the associations between SNPs or haplotype of the genes and milk production traits, SCS were analyzed by the least squares method in the GLM procedure of SAS. The IL-23R c.-856A>G and IL-23R c.-207T>C showed close linkage disequilibrium ( r 2 =0.89). No association was found with SCS, but associations were found between 3 of these SNP with milk protein content and lactose content. The software MatInspector revealed that these SNPs were located within several potential transcription factor binding sites, and may alter gene expression, but further investigation will be required to elucidate the biological and practical relevance of these SNP. • One new SNP( c.-246G>T ) in the promoter of IL-12Rβ2 gene was identified. • The potential TFBS were analyzed for the promoter of IL-12Rβ2 and IL-23R genes. • The IL-23R c.-856A>G and IL-23R c.-207T>C showed close LD ( r 2 =0.89). • No association was found between 3 SNPs and SCS.

Prenatal Nicotinic Exposure (PNE) Promotes Pulmonary IL‐1β and 5‐HT and Gene/Protein Expression of IL1R1, NK1R, ADRA1, and TRPV1 in N/J ganglia Neurons via Acting on α7nAChRs in Rat Pups

We have shown that PNE sensitizes bronchopulmonary C‐fibers (PCFs) in rat pups. Here we asked if PNE was able to increase the pulmonary mediators known to sensitize/active PCFs, and their corresponding receptors expressed in nodose/jugular (N/J) ganglion neurons in P11‐14 pups. If so, we further asked if the evoked changes depended on nicotinic acetylcholine receptors (nAChRs), particularly α7nAChR. We compared IL‐1β, substance P (SP), serotonin (5‐HT), and adenosine (AD) in BALF and IL‐1 type I receptor (IL1R1), NK1R, TRPV1, and adenosine A1 receptor (ADRA1) in the N/J ganglia in P11‐14 rat pups prenatally exposed to saline (Ctrl) and PNE alone or coupled with mecamylamine (MM), a non‐selective blockade of nAChRs and methyllycaconitine (MLA), a selective blockade of α7nAChR. We found that PNE significantly augmented IL‐1β and 5‐HT but not SP and AD in BALF and gene and protein expressions of IL1R1, NK1R, TRPV1, and ADRA1 in the N/J ganglia. Both MM and MLA abolished the responses of IL‐1β and 5‐HT in BALF. MM eliminated the gene and protein responses of N/J ganglion TRPV1, NK1R, IL1R1, and ADRA1 to PNE except little effect on IL1R1 gene, while MLA eliminated all of the gene responses. Our data suggest that PNE is able to elevate IL1R1, NK1R, ADRA1, and TRPV1 in the N/J ganglia, which, along with increased IL‐1β and 5‐HT releases in the lungs, likely contributes to sensitize/activate PCFs. These PNE‐induced modulations are greatly dependent on the activation of α7nAChRs (Supported by HL‐107462).