Gene – environment treatment and asthma

printing supported by . Visit Chiesi at Stand B2.10 SUNDAY, SEPTEMBER 2ND 2012 Conclusions: TSLP variants are significantly associated with bronchial asthma. TSLP might be a new therapeutic target molecule for asthma. P471 Association of TGFB1 and IL4RnG2 gene polymorphisms with asthma Pedro J. Romero Palacios1, Juan Carlos Alavrez Merino2, Carolina Cisneros Serrano3, Pablo De Diego Fernández4, Francisco J. González Barcala5, José María Ignacio García6, Antonio Pereira Vega7, Luis Pérez de LLano8, Isabel Urrutia Landa9. 1Medicina, Facultad de Medicina. Universidad de Granada, Granada, Andalucía, Spain; 2Medicina Legal, Toxicología y Psiquiatría, Facultad de Medicina. Universidad de Granada, Granada, Andalucía, Spain; 3Servicio de Neumología., Hospital Universitario de la Princesa, Madrid, Spain; 4Servicio de Pediatría, Hospital Universitario Virgen de las Nieves, Granada, Spain; 5Servicio de Neumología, Hospital de Loureiro Crespo, Pontevedra, Galicia, Spain; 6Servicio de Neumología, Hospital Serranía de Ronda, Málaga., Andalucía, Spain; 7Servicio de Neumología, Hospital Juan Ramón Jiménez, Huelva, Andalucía, Spain; 8Servicio de Neumología, Hospital Lucus Augusti, Lugo, Galicia, Spain; 9Servicio de Neumología, Hospital de Galdakao, Bixkaia, Pais Vasco, Spain There are components of the asthma phenotype that appear to be strongly heritable. We have studied certain SNP variations in a set of genes related to the pathogenesis and evolution of asthma in patients with severe asthma and their immediate relatives, as well as in a control group of non-asthmatics patients. Aims of this study: 1. To determine the existence of common genetic characteristics in individuals suffering from severe asthma. 2. To study whether there are common genetic patterns in relatives of patients with severe asthma. Method: We selected patients diagnosed of severe persistent asthma according to the criteria proposed by the ATS Consensus for Definition of Severe/Refractory Asthma and their fist-line relatives, symptomatic or non-symptomatic according to their responses to the European Community Respiratory Health Survey Questionnaire. A control group of non-asthmatic patients was also included. We obtained saliva samples to study 10 different SNPs located on different genes related to asthma (CHI3L1: rs4950928; ADRB2: rs1042713, rs1042714; DENND1B: rs1775456; ORMDL3: rs7216389; TMCO6: rs2569190; ADAM33: rs2280091; IL4: rs2243250; TGFB1: rs1800469; IL4R: rs1801275), by TaqMan SNP Genotyping Assay. Results: We analyzed samples from 150 patients diagnosed of severe persistent asthma, 49 controls, 69 asymptomatic relatives and 83 symptomatic relatives. We found significant differences (p <0.05) between the group of patients and relatives with respect to controls in two pairs of the alleles studied: TGFB1 rs1800469; IL4RnG2: rs1801275. Conclusions: We found a higher prevalence of TGFB1 rs1800469 (AA and AG) in patients with severe asthma and their relatives; and IL4RnG2: rs1801275 (AA) in asthmatics. P472 Investigating the role of IREB2 genetic variants in susceptibility to COPD Noor Kalsheker1, Sally Chappell1, Tamar Guetta-Baranes1, Aiman Alsaegh1,2. 1Molecular Medical Sciences, The University of Nottingham, United Kingdom; 2Faculty of Applied Medical Science, The Uinversity of Umm Alqura, Makkah, Saudi Arabia The IREB2 gene encodes the iron-binding protein 2, which is a major regulator of iron homeostasis. Several studies have shown that the IREB2 locus is a contributor to COPD susceptibility. Previously, we observed significant associations of seven IREB2 genetic variants with increased risk of COPD in a large case-control study. Subsequent in-silico analysis showed that six of these SNPs were in tight linkage disequilibrium with two variants that lie within the promoter (rs2656070) and the 3’UTR (rs12899351) of IREB2 gene. The promoter SNP is predicted to disrupt the binding of two transcription factors while the 3’UTR SNP is located in a region that is predicted to be a target site of mir-1285 and mir-5096. The aims of this study were to evaluate the functional effect of these variants on IREB2 expression. To test the effect of the promoter SNP, two fragments (one for each allele) of the 5’region of IREB2 were inserted upstream of the luciferase gene in the pGL3-Basic vector and then transfected into the A549 cells. Our results show that there was no difference in luciferase expression from cells transfected with rs2656070 wild type construct compared with the risk allele under basal conditions. Further analysis will be undertaken to examine the effect of the rs2656070 under different stimulatory conditions. For the 3’UTR SNP, two fragments (for both alleles) spanning the potential miRNA target site was cloned downstream of the luciferase gene in pmirGLO vector and then transfected into HepG2 cells that are known to express mir-1285. Luciferase assay showed that mir-1285 did not recognize the cloned sequence. Additional investigation will consider the regulatory role of mir-5096 on IREB2 expression. P473 Case-control association analysis of candidate genes in asthma, rhinitis and COPD: A preliminary report Cristina Bombieri1, Anna Rita Lo Presti1, Francesca Belpinati1, Simone Accordini2, Alessandro Baldan1, Marcello Ferrari3 , Giovanni Malerba1, Elisabetta Zanolin2, Pier Franco Pignatti1 , Roberto de Marco2. 1Dpt. of Life and Reproduction Sciences, Section of Biology and Genetics, University of Verona, Italy; 2Dpt. of Medicine and Public Health, Section of Epidemiology and Medical Statistics, University of Verona, Italy; 3Dpt. of Medicine, Section of Internal Medicine D, University of Verona, Italy This study aims to determine the genetic involvement in the susceptibility to asthma, rhinitis and COPD, by candidate gene association analysis, in a large and accurately defined series of Italian subjects, even considering exposure to some environmental contexts and life-styles. The study population included 1075 subjects (aged 20-66 years) from the general population, enrolled in the frame of the Gene Environment Interactions in Respiratory Diseases (GEIRD) study between 2007 and 2010. Cases and controls were diagnosed during a clinical examination that included a detailed interview, pre/post bronchodilator spirometry, methacoline challenge, skin prick tests. A panel of 384 Single Nucleotide Polymorphisms (Tag-SNP), representative of 63 candidate genes with a previous indication of possible association to the studied diseases, was genotyped by a customized GoldenGate Genotyping assay. Presently, genotyping of 725/1075 subjects are completed. A preliminary association study of candidate gene polymorphisms was conducted on these data, for the susceptibility to one or more of the studied phenotypes, by basic association test based on allele frequency comparison. Presence of association (unadjusted p<0.005) was observed between GSTP1 and non-atopic rhinitis, PDE4D and ever asthma with atopy, IL13 and past-asthma, TNS1 and chronic bronchitis. Moreover, a possible association (unadjusted p<0.02) was also found for IL1RL2 with ever asthma, chronic bronchitis, atopic rhinitis and non atopic rhinitis. The analysis is going on to complete the genotyping of all the enrolled subjects and to perform haplotype analysis, to confirm the involvement of these genes in the studied diseases. P474 Association analysis of β2-adrenergic receptor gene polymorphisms (Arg16Gly and Gln27Glu) with asthma in the Volga-Ural region of Russia Yuliya Fedorova1, Alexandra Karunas1, Galiya Gimalova1, Nailya Ramazanova2, Regina Murzina2, Lilya Muchtarova3, Elena Galimova3, Radik Gatiyatullin2, Shamil Zagidullin3, Esfir Etkina2, Elza Khusnutdinova1. 1Department of Human Molecular Genetics, Institute of Biochemistry and Genetics, Ufa Scientific Centre of RAS, Ufa, Russian Federation; 2Department of Pediatrics, Bashkir State Medical University, Ufa, Russian Federation; 3Department of Propaedeutics of Internal Diseases, Bashkir State Medical University, Ufa, Russian Federation The β2-adrenergic agonists are the most potent bronchodilators for the treatment of asthma. Genetic variation in the ADRB2 gene has been hypothesized to have a role in differential response to beta-agonist (BA) therapy in asthma. Two polymorphic variants rs1042713 (Gly16Arg) and rs1042714 (Gln27Glu) were genotyped in 618 patients with physician-diagnosed asthma, aged 2-60 years (192 Russians, 139 Tatars, 82 Bashkirs and 205 mixed origins), and 366 nonasthmatic individuals (123 Russians, 91 Tatars, 51 Bashkirs and 101 mixed origins) from the Volga-Ural region of Russia. Genotypes were determined by the PCR-RFLP method. Data were analyzed using the chi-square test with Haploview software. We found significant association of Gln27Glu polymorphism with mild decrease FEV1 (60-80% of the predicted value) in Russian patients. The frequencies of genotype ADRB2*27Gln/27Gln and allele ADRB2*27Gln were increased in asthmatics of this group compared to controls (p=0,03, OR=2,25 (95%CI 1,05-4,82) and p=0,02, OR=1,97 (95%CI 1,11-3,50), accordingly). The analysis of Arg16Gly polymorphism showed significant association with moderate asthma in Tatars. The higher frequency of the ADRB2*16Gly/16Arg heterozygotes was revealed in the patients compared with controls (p=0,02, OR=2,25 (95%CI 1,15-4,03)). In summary, these results suggest an important role for polymorphisms of gene ADRB2 in the development of asthma in the Volga-Ural region of Russia. 65s Poster Discussion Room C7 10:45 12:45 Abstract printing supported by . Visit Chiesi at Stand B2.10printing supported by . Visit Chiesi at Stand B2.10 SUNDAY, SEPTEMBER 2ND 2012 P475 Gene polymorphisms, gene expression and inflammatory markers in preschool children with and without wheeze Ester Klaassen1, Kim van de Kant1, Quirijn Jobsis1, John Penders2, Frederik Jan van Schooten3, Marieke Quaak4, Gertjan den Hartog3, Gerard Koppelman5, Constant van Schayck4, Guillaume van Eys6, Edw