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NASAL EPITHELIUM GENE EXPRESSION PROFILING IN CHILD RESPIRATORY ALLERGIC DISEASE
YOUNG INVESTIGATOR’S PRESENTATIONS
while pretreatment with steroids did not suppress IL-8 upregulation. Pretreatment with GSH-OEt prior to ozone and steroid exposure suppressed changes in intracellular oxidative state, as well as the increases in IL-8 and also IL-6. Ozone exposure increased the transcription factor, NF-xB; this change was suppressed by steroids but suppressed further by steroid plus GSH-OEt pretreatment.
CONCLUSION Steroids suppress NF-xB and, in turn, pro-inflammation cytokines. These agents also further disturb the intracellular glutathione redox state in ozone-exposed airway epithelial cells activating pro-inflammatory signaling via NF-xB. Giving antioxidants to steroid related oxidative stress could induce more effectiveness of oral steroid therapy in airway diseases.
S271
D17S959. This critical region spans approximately 6 megabases of genomic DNA. Work is in progress to refine the critical region by identification of allelic association. One of the 20 known genes in this region, Tektin 3 encodes the filamental protein tektin known to be an integral component of cilia and sperm flagella. Tektin 2 (another protein from same gene family) knock out mice model exibits phenotype similar to that of human PCD. The gene is expressed in the lungs and testes and has been shown to play a role in lung morphogenesis. Tektin 3 therefore represents an excellent candidate gene for PCD. Its genomic characterisation will be undertaken before performing mutational analysis in patients. Correspondance to Dr R Chodhari, Wellcome Trust Reserch Fellow. [email protected] DOI: 10.1016/j.prrv.2006.04.151
DOI: 10.1016/j.prrv.2006.04.139
F15/125 – Identification of a novel locus for primary ciliary dyskinesia (PCD) with inner and outer dyenin arm defects in the Pakistani population R. Chodhari1, C. O’Callaghan2, E. Moya3, J. Dankert-Roelse4, G. Pals4, R.M. Gardiner1, E.M.K. Chung1 and H.M. Mitchison1 1 University College London, Department of Paediatrics and Child Health, University College London, London, UK; 2 University of Leicester, Division of Child Health, Leicester, UK; 3Bradford Royal Infirmary Hospital, Bradford, UK; 4Vrije Universiteit Medical Centre, Department of Paediatrics, Amesterdam, UK PCD is an autosomal recessive disorder characterised by respiratory tract infections, sinusitis, bronchiectasis and subfertility. It affects 1:20,000 live births. The clinical phenotype results from dysmotility of the cilia, which is associated with a variety of ultrastructural abnormalities. The core or axoneme of cilia comprises a bundle of microtubules and associated proteins including dyneins, nexin links and radial spokes. About 50% of patients exhibit laterality defects, commonly situs inversus, an association classed as Kartagener syndrome. The relationship between ciliary defects and abnormalities of left-right axis determination remains unknown. We have studied five families with a ciliary defect of absent inner and outer dynein arms from the Mirpuri population of northern Pakistan. All the parents are first cousins and there are a total of nine affected (eight of whom have situs inversus) and twentynine unaffected individuals. A 400 marker microsatellite genome wide scan identified a region consistent with linkage in three of the five families on chromosome 17p12. Using GENEHUNTER, a maximum multipoint lod score of 3.9 was obtained between D17S799 and
F16/302 – Nasal epithelium gene expression profiling in child respiratory allergic disease L. Giovannini-Chami1,2,3, P. Rostagno2,3, V. Virolle2,3, L. Castillo1, J.F. Michiels4, T. Bourrier1, D. Crenesse5, M. Albertini1 and P. Barbry2,3 1 Department of Pediatrics, Unit of Pneumology and Allergology, Hopital de l’Archet, Nice, France; 2CNRS, Institut de Pharmacologie Mole´culaire et Cellulaire, UMR 6097, 660, routes des Lucioles F-06560, Sophia-Antipolis, France; 3 University of Nice Sophia Antipolis, Institut de Pharmacologie Mole´culaire et Cellulaire, UMR 6097, 660, routes des Lucioles F-06560, Sophia-Antipolis, France; 4Department of Anatomopathology, Hopital de l’Archet, Nice, France; 5 Department of Pediatric Functional Explorations, Hopital de l’Archet, Nice, France
INTRODUCTION Asthma is the most frequent chronic disease in childhood. Allergic rhinitis has been described as a risk factor to develop asthma. The objective of the study was to evaluate the contribution of the respiratory epithelium to development of allergic rhinitis and asthma and to identify the molecular mechanisms driving rhinitis toward asthma.
METHODS Using a human pangenomic microarray, we established expression profiles of nasal epithelial cells, collected by brushing of patients belonging to one of four distinct groups: (a) allergic rhinitis to dust mite (AR) isolated (n = 12), (b) AR associated with bronchial hyperreactivity (n = 12), (c) AR associated with asthma (n = 14), (d) control (n = 14).
S272
YOUNG INVESTIGATOR’S PRESENTATIONS
RESULTS
CONCLUSION
Among 188 genes statistically modulated in the nasal epithelium between asthmatics and controls, 42 are located on asthma loci of susceptibility and 33 have already been linked to asthma in literature. A supervised analysis determined a set of 16 genes (CST4, ALOX15, CLCA1. . .) allowing the correct classification of 11 asthmatics out of 14 and 13 controls out of 14. However, when this classifier was used to predict cases of rhinitis without asthma, a bimodal distribution was observed, with subjects approaching asthmatic patients phenotype and others approaching control phenotype. This classification was independent of the status of bronchial hyperreactivity.
The technique developed here allows pangenomic analysis of a small number of cells (500 000). It is well adapted for minimally invasive investigations on patients with wellcharacterized clinical profiles. This analysis confirms the central place of respiratory epithelium in asthmatic airway remodeling and identifies an epithelial phenotype close to asthmatic ones in patients who only have a rhinitis, independently of their bronchial hyperreactivity status. An ongoing clinical follow-up will evaluate the capacity of our classifier to predict evolution toward asthma. DOI: 10.1016/j.prrv.2006.04.152
while pretreatment with steroids did not suppress IL-8 upregulation. Pretreatment with GSH-OEt prior to ozone and steroid exposure suppressed changes in intracellular oxidative state, as well as the increases in IL-8 and also IL-6. Ozone exposure increased the transcription factor, NF-xB; this change was suppressed by steroids but suppressed further by steroid plus GSH-OEt pretreatment.
CONCLUSION Steroids suppress NF-xB and, in turn, pro-inflammation cytokines. These agents also further disturb the intracellular glutathione redox state in ozone-exposed airway epithelial cells activating pro-inflammatory signaling via NF-xB. Giving antioxidants to steroid related oxidative stress could induce more effectiveness of oral steroid therapy in airway diseases.
S271
D17S959. This critical region spans approximately 6 megabases of genomic DNA. Work is in progress to refine the critical region by identification of allelic association. One of the 20 known genes in this region, Tektin 3 encodes the filamental protein tektin known to be an integral component of cilia and sperm flagella. Tektin 2 (another protein from same gene family) knock out mice model exibits phenotype similar to that of human PCD. The gene is expressed in the lungs and testes and has been shown to play a role in lung morphogenesis. Tektin 3 therefore represents an excellent candidate gene for PCD. Its genomic characterisation will be undertaken before performing mutational analysis in patients. Correspondance to Dr R Chodhari, Wellcome Trust Reserch Fellow. [email protected] DOI: 10.1016/j.prrv.2006.04.151
DOI: 10.1016/j.prrv.2006.04.139
F15/125 – Identification of a novel locus for primary ciliary dyskinesia (PCD) with inner and outer dyenin arm defects in the Pakistani population R. Chodhari1, C. O’Callaghan2, E. Moya3, J. Dankert-Roelse4, G. Pals4, R.M. Gardiner1, E.M.K. Chung1 and H.M. Mitchison1 1 University College London, Department of Paediatrics and Child Health, University College London, London, UK; 2 University of Leicester, Division of Child Health, Leicester, UK; 3Bradford Royal Infirmary Hospital, Bradford, UK; 4Vrije Universiteit Medical Centre, Department of Paediatrics, Amesterdam, UK PCD is an autosomal recessive disorder characterised by respiratory tract infections, sinusitis, bronchiectasis and subfertility. It affects 1:20,000 live births. The clinical phenotype results from dysmotility of the cilia, which is associated with a variety of ultrastructural abnormalities. The core or axoneme of cilia comprises a bundle of microtubules and associated proteins including dyneins, nexin links and radial spokes. About 50% of patients exhibit laterality defects, commonly situs inversus, an association classed as Kartagener syndrome. The relationship between ciliary defects and abnormalities of left-right axis determination remains unknown. We have studied five families with a ciliary defect of absent inner and outer dynein arms from the Mirpuri population of northern Pakistan. All the parents are first cousins and there are a total of nine affected (eight of whom have situs inversus) and twentynine unaffected individuals. A 400 marker microsatellite genome wide scan identified a region consistent with linkage in three of the five families on chromosome 17p12. Using GENEHUNTER, a maximum multipoint lod score of 3.9 was obtained between D17S799 and
F16/302 – Nasal epithelium gene expression profiling in child respiratory allergic disease L. Giovannini-Chami1,2,3, P. Rostagno2,3, V. Virolle2,3, L. Castillo1, J.F. Michiels4, T. Bourrier1, D. Crenesse5, M. Albertini1 and P. Barbry2,3 1 Department of Pediatrics, Unit of Pneumology and Allergology, Hopital de l’Archet, Nice, France; 2CNRS, Institut de Pharmacologie Mole´culaire et Cellulaire, UMR 6097, 660, routes des Lucioles F-06560, Sophia-Antipolis, France; 3 University of Nice Sophia Antipolis, Institut de Pharmacologie Mole´culaire et Cellulaire, UMR 6097, 660, routes des Lucioles F-06560, Sophia-Antipolis, France; 4Department of Anatomopathology, Hopital de l’Archet, Nice, France; 5 Department of Pediatric Functional Explorations, Hopital de l’Archet, Nice, France
INTRODUCTION Asthma is the most frequent chronic disease in childhood. Allergic rhinitis has been described as a risk factor to develop asthma. The objective of the study was to evaluate the contribution of the respiratory epithelium to development of allergic rhinitis and asthma and to identify the molecular mechanisms driving rhinitis toward asthma.
METHODS Using a human pangenomic microarray, we established expression profiles of nasal epithelial cells, collected by brushing of patients belonging to one of four distinct groups: (a) allergic rhinitis to dust mite (AR) isolated (n = 12), (b) AR associated with bronchial hyperreactivity (n = 12), (c) AR associated with asthma (n = 14), (d) control (n = 14).
S272
YOUNG INVESTIGATOR’S PRESENTATIONS
RESULTS
CONCLUSION
Among 188 genes statistically modulated in the nasal epithelium between asthmatics and controls, 42 are located on asthma loci of susceptibility and 33 have already been linked to asthma in literature. A supervised analysis determined a set of 16 genes (CST4, ALOX15, CLCA1. . .) allowing the correct classification of 11 asthmatics out of 14 and 13 controls out of 14. However, when this classifier was used to predict cases of rhinitis without asthma, a bimodal distribution was observed, with subjects approaching asthmatic patients phenotype and others approaching control phenotype. This classification was independent of the status of bronchial hyperreactivity.
The technique developed here allows pangenomic analysis of a small number of cells (500 000). It is well adapted for minimally invasive investigations on patients with wellcharacterized clinical profiles. This analysis confirms the central place of respiratory epithelium in asthmatic airway remodeling and identifies an epithelial phenotype close to asthmatic ones in patients who only have a rhinitis, independently of their bronchial hyperreactivity status. An ongoing clinical follow-up will evaluate the capacity of our classifier to predict evolution toward asthma. DOI: 10.1016/j.prrv.2006.04.152