1. AllerGen Workshop, Vancouver March 2009 - FK -1 The Egea study Francine Kauffmann Respiratory and Environmental Epidemiology Inserm, U780 - Epidemiology and Biostatistics Villejuif, France Genes and the Environment: The Genesis of Asthma and Allergy Workshop March 1-3, 2009 - Vancouver

2. AllerGen Workshop, Vancouver March 2009 - FK -2 General objectives 1)To determine genetic factors of asthma and related traits (atopy and bronchial responsiveness) 2)To determine environmental factors and the interactions of these 2 types of factors 3) To clarify the heterogenetiy of asthma

3. AllerGen Workshop, Vancouver March 2009 - FK -3 Question => Design Genetics Environment Heterogeneity Families of asthmatic cases Segregation Linkage Association Detailed data on phenotypes => Asthmas Large nb of « usual » asthma nuclear families Prevalence => hospital recruitment Detailed data on environment (personal, familial, general, childhood, adulthood) Controls. Further, interest to estimate the allelic frequency of genetic markers and for association studies Clin Experim Allergy 1995

4. AllerGen Workshop, Vancouver March 2009 - FK -4 Egea - A multicentric study

5. AllerGen Workshop, Vancouver March 2009 - FK -5 Case-control study - 5 cities in France + Family study of asthmatic probands + families with affected sibs =>In total 2105 subjects

6. AllerGen Workshop, Vancouver March 2009 - FK -6 A longitudinal study Survey12New at Egea2 Brief self-completed Q (step 1) and full questionnaire by an interviewer (step 2) Respiratory and allergic symptoms (ECRHS- like + complements) ++New aspects ECRHS 2 Drug consumption++3 and 12 last months; dose of steroids Quality of life (SF36, AQLQ)-+ Skin prick tests 11 ag (ECRHS+2)+++ cypress LFT, methacholine challenge (4mg), WBC, IgE++ Exhaled NO, Exhaled breath condensate-+FeNO (3 centers), EBC Blood (serum, plasma, DNA, cell lines)++Strict conditions + RNA + erythrocytes Now DNA for 97% - RNA for 66% Environment Smoking, occupational exposures, indoor exposures, pets, air pollution ++Complete ccupational and residential histories, cleaning products, window of exposures for pets Country living+Detailed egea2 Diet, physical activity,-+Silhouettes, FFQ (118 items) Hormone-related events-+ECRHS questionnaire

7. AllerGen Workshop, Vancouver March 2009 - FK -7 First survey Egea1 N = 2047 Dead N = 45 (2.2%) Only short questionnaire (step1) N = 302 (15.1%) Long questionnaire (step 2) N = 1543 (77.1 %) Alive N = 2002 Refusals and lost N= 157 (7.8%)Overall follow-up N = 1845 (92.2%)..with a good follow-up

8. AllerGen Workshop, Vancouver March 2009 - FK -8 In summary Case-control + family - 2105 subjects (388 asthmatic families + 415 controls), 7- 70 years in 1992, French multicentric Longitudinal - Egea1 1992/5 - Egea2 2003/7 (77% with exam, 92% with simple info) Health data (respiratory and allergy) (ECRHS-like) asthma - rhinitis - eczema - lung function - allergic tests ~1500 var x 2 Environment : smoking, occupational and residential histories, indoor exposure, fam animals and pets, job exposure matrix/ expert, air pollution (FP7 Escape ; PI B Brunekreef), diet, physical activity Genetic program (DNA 97%) 400 µsat + 3500 snps + GWA 550K snps (FD coord, CNG ; FP6 Gabriel PI B Cookson) Biological collection : RNA 1400 (egea2) + aliquotes serum, plasma, erythrocytes or exhaled breath condensate 70 000 (1/5 egea1, 4/5 egea2) Egea collection 2105 Founders/offspring 897/1207 Asthma,ever 2104 [99.9%] N (%) 902 (43%) DNA 2034 [97%] RNA 1398 [66%] Serum Egea1 1953 [93%] Serum Egea2 1427 [68%] E xhaled B reath C ondensate Egea2 1336 [63%]

9. AllerGen Workshop, Vancouver March 2009 - FK -9 Among potential areas of international collaborations Environment Genetics

10. AllerGen Workshop, Vancouver March 2009 - FK -10 Occupation - Egea Environmental factor usually not shared by family members Complete occupational history - All jobs coded Description of tasks, used to improve evaluation of exposures by the Asthma Job exposure matrix* to asthmogens through expert step Detailed information regarding exposures for specific jobs (cleaning, nurses,..) with interest in cleaning and disinfecting agents Information on health-related changes in jobs *Kennedy et al Asthma JEM (OEM), described in Egea in 2000, now internationally used.

11. AllerGen Workshop, Vancouver March 2009 - FK -11 Air pollution - Egea Environmental factor usually shared by family members Complete residential history : address egea1 et ege2 (all geocoded), commune all life (birth included) Exposure modelled estimates available for France –Géovariances (French map 4 km 2 NO 2, SO 2, O 3 and PM 10 (models - cokriging, i.e interpolation between fixed stations improved by considering local land characteristics) - Publications Rage et al.2008/9 –APMoSHERE (European map based on 1 km 2 D Briggs) 2001 NO 2 and O 3 European collaboration - B Jacquemin ECRHS –Futur Escape (EU FP7) - Land use regression for specific locations covering most of Egea - Evaluations available in 2011

12. AllerGen Workshop, Vancouver March 2009 - FK -12 Genetics - Egea A program which started in 1994 - Microsatellites and SNPS - Genome wide linkage analyses - Candidate genes - Numerous publications Part of Gabriel Interest in candidate gene studies, in genome wide approaches, with methodological developments, in gene environment interactions, in case-control comparisons, in familial data analyses,..

13. AllerGen Workshop, Vancouver March 2009 - FK -13 p<10 -5 Exposed Unexposed Risk of asthma before age 4 Age of Onset of Asthma & Env Tobacco Smoke Exposure Bouzigon et al, New Engl J Med, 2008  risk of early-onset asthma with ETS exposure 17q21 locus associated with early-onset asthma (  4 yrs) Asthma onset  4 yrs  Exploration of genetic heterogeneity according to age of onset of asthma in regions detected by meta-analysis of asthma GWAS  Genome-wide association analysis of age of onset of asthma  Investigation of G x Exposure to passive smoking in this context

14. AllerGen Workshop, Vancouver March 2009 - FK -14 PARENT-OF-ORIGIN EFFECT / IMPRINTING Pooled genome-wide linkage analysis of French, Italian and UK Family data has investigated parent-of-origin effects for asthma, skin tests, IgE Malerba, Bouzigon et al, GABRIEL exchange programme 3 teams: France (F Demenais), Italy (P Pignatti), UK (W Cookson) Several regions show parent-of-origin effects for asthma, SPT, IgE Proposal: GWAS to investigate imprinting in pooled family data sets

15. AllerGen Workshop, Vancouver March 2009 - FK -15 GENES WITH PLEIOTROPIC EFFECTS Previous EGEA linkage scans have characterized regions of linkage common to several asthma-related phenotypes, suggesting pleoitropic effect of genes. Some of these linkages are sex-specific & show Gxsmoking interactions (Bouzigon et al, Hum Mol Genet, 2004, Bouzigon et al, Hum Genet, 2007; Aschard et al, submitted) Proposal: GWAS for identifying pleiotropic genes influencing asthma-related phenotypes (eg asthma onset, FEV1, Skin tests) possibly taking into account GXE ( smoking, gender…)

16. AllerGen Workshop, Vancouver March 2009 - FK -16 Strengths - Egea Families and population-based controls Phenotypic characterisation (detailed and longitudinal) Environment Biological collection

17. AllerGen Workshop, Vancouver March 2009 - FK -17 Limitations - Egea Complexity of design - numerous strategies (internal replication, but complex) need to learn how to work with the study - post doc Sample size - 2000 not 20 000, but 902 asthmatics, precision (power) Limited biological data evaluating environment - but validated measures are few Funding insufficient for maintaining data sets, web site,.. (infrastructure), statistical analyses,… and post doc

18. AllerGen Workshop, Vancouver March 2009 - FK -18 Coordination : F Kauffmann; F Demenais (genetics); I Pin (clinical aspects). Respiratory epidemiology : Inserm U 700, Paris M Korobaeff (Egea1), F Neukirch (Egea1); Inserm U 707, Paris : I Annesi-Maesano ; Inserm U 780, Villejuif : F Kauffmann, N Le Moual, R Nadif, MP Oryszczyn ; Inserm U 823, Grenoble : V Siroux Genetics : Inserm U 393, Paris : J Feingold ; Inserm U 535, Villejuif : MH Dizier ; Inserm U 946, Paris : E Bouzigon, F Demenais ; CNG, Evry : I Gut, M Lathrop. Clinical centers : Grenoble : I Pin, C Pison; Lyon : D Ecochard (Egea1), F Gormand, Y Pacheco ; Marseille : D Charpin (Egea1), D Vervloet ; Montpellier : J Bousquet ; Paris Cochin : A Lockhart (Egea1), R Matran (now in Lille) ; Paris Necker : E Paty, P Scheinmann ; Paris-Trousseau : A Grimfeld, J Just. Data and quality management : Inserm ex-U155 (Egea1) : J Hochez ; Inserm U 780, Villejuif : N Le Moual, C Ravault ; Inserm ex-U 794 : N Chateigner ; Grenoble : J Ferran Egea cooperative group

19. AllerGen Workshop, Vancouver March 2009 - FK -19 For further information,.. Web site of the study (design, questionnaires, list of publications and communications) http://ifr69.vjf.inserm.fr/~egeanet/ As Canada is a bilingual country, please note that All abstracts of papers are in English and French on egeanet Articles de synthèse en français (données Egea1) Siroux V, et coll. Rev Mal Respir 2007 ; 24 : 599-608 (soon in English,..) Kauffmann F, et coll. Rev Mal Respir 2002 ;19:63-72