1. Predisposition to asthma among the Utah population Craig Teerlink University of Utah Department of Biomedical Informatics Asthma Genomics Conference Utah Department of Health Asthma Program June 7, 2007

2. Introduction Asthma is a common disorder Effects 7% of US population 1 Effects 7% of US population 1 Increased global incidence observed in the last few decades 2 Increased global incidence observed in the last few decades 2 Complicated etiology Both environmental and genetic factors are recognized 3 Both environmental and genetic factors are recognized 3 Heritable nature of asthma Heritable nature of asthma Discordance observed between MZ and DZ twins 4 Familial aggregation studies and risk factor analyses provide evidence that asthma clusters in families 5,6 A better understanding of predisposing factors may help improve treatment outcomes

3. Introduction to familial analysis study Such studies have been restricted to first degree relatives It is difficult to distinguish between evidence for common genetic factors and common environmental factors among close relatives since close relatives often share their immediate environment In contrast, using a unique Utah resource, we were able to observe increased risk to distant relatives for a severe asthma phenotype definition 7

4. The Utah Population Database Computerized genealogy records 2.2 million Utah pioneers and their descendents Some genealogies have up to 10 generations Has been linked to 440,000+ death certificates from Utah The combined resource allows us to identify individuals who died from asthma (cases) and investigate their ‘relatedness’

5. Benefits of genealogical approach to familiality Well-established methods The resource has previously been used to provide evidence for a heritable component in other disease settings The resource has previously been used to provide evidence for a heritable component in other disease settings Can extend analyses to distant relatives (i.e., 2 nd or 3 rd degree relatives), providing potentially more meaningful results

6. Two types of analysis Relative risk If asthma mortality is familial, a higher risk of asthma mortality will be found among relatives of individuals who died from asthma than would be found for random controls If asthma mortality is familial, a higher risk of asthma mortality will be found among relatives of individuals who died from asthma than would be found for random controls Average relatedness If asthma mortality is familial, more relationships between cases will be found than would be found for random controls If asthma mortality is familial, more relationships between cases will be found than would be found for random controls

7. Relative risk analysis Method Compare the rate of asthma death in relatives of asthma death cases with the rate of asthma death in the population (UPDB) Compare the rate of asthma death in relatives of asthma death cases with the rate of asthma death in the population (UPDB) Results for 1,553 asthma deaths No. of relatives ObservedExpectedRRP-value 1 st degree relatives 7,9365230.71.69<0.001 2 nd degree relatives 19,31910074.81.340.003 3 rd degree relatives 28,601129112.21.150.065

8. Average relatedness analysis Method Calculate the genetic distance between every pair of cases (i.e., degree of ‘relatedness’) Calculate the genetic distance between every pair of cases (i.e., degree of ‘relatedness’) Calculate the average relatedness of all cases (GIF statistic) Calculate the average relatedness of all cases (GIF statistic) Repeat for 1000 sets of matched controls Repeat for 1000 sets of matched controls Results for 1,553 asthma deaths All cases Ignoring 1 st and 2 nd degree relatives Case GIF 3.161.95 Control GIF 2.421.73 P-Value<0.00010.026

9. Contribution to the GIF statistic Contribution to the GIF statistic by genetic distance between pairs of individuals for asthma mortality 1,553 cases and 1000 sets of matched controls

10. Summary of familial investigation Used a population based genealogy linked to death certificates Observed significantly increased risk to relatives of individuals who died from asthma Cases are significantly more related than expected by chance Both analyses were significant in close and distant relatives

11. Implications Implications vary according to interest… Genetic epidemiologist: Highly specific phenotype definition and significant results among distant relatives suggests heritable factor Highly specific phenotype definition and significant results among distant relatives suggests heritable factor Department of health: Risk estimates are on a population basis, so apply well to an entire population Risk estimates are on a population basis, so apply well to an entire population An individual: Asthma mortality is rare Asthma mortality is rare Increased risk is low and not likely to apply at the individual level Increased risk is low and not likely to apply at the individual level

12. Next step Use of clinical data (instead of mortality) to distinguish asthma cases within the genealogy database may produce more meaningful risk estimates to clinicians, public health practitioners, and individuals. Utah Asthma Program community mini- grant may help to perform the next step

13. Acknowledgements, 1 People Lisa Cannon-Albright Lisa Cannon-Albright Matt Hegewald Matt HegewaldInstitutions Resource for Genetic and Epidemiologic Research (Utah Population Database) Resource for Genetic and Epidemiologic Research (Utah Population Database) Utah Department of Health Asthma Program Utah Department of Health Asthma Program

14. Introduction to linkage analysis study Linkage analysis Attempts to identify disease predisposition loci in the genome Attempts to identify disease predisposition loci in the genome Based on the phenomenon of chromosome recombination that occur during meioses Based on the phenomenon of chromosome recombination that occur during meioses Utilizes inheritance information gathered in disease pedigrees Utilizes inheritance information gathered in disease pedigrees Previous genome-wide scans for asthma have implicated almost every chromosome 22 study populations thus far 8 22 study populations thus far 8 > 30 suggestive or significant regions in the genome 8 > 30 suggestive or significant regions in the genome 8 Several genes have been identified/hypothesized in association studies 9 Replication is needed for these genes Replication is needed for these genes Results are likely to be population-specific Results are likely to be population-specific

15. 1 2 3 4 5 6 7 8 9101112 13 14 15 16 17 18 1920 2122 X Previous results from genome-wide scans for asthma 8 previously published regions

16. A unique data resource for asthma linkage 81 extended pedigrees ascertained for asthma between 1996 and 2000 3 to 6 generations per pedigree 3 to 6 generations per pedigree 6 to 97 individuals per pedigree 6 to 97 individuals per pedigree 2 to 40 affected individuals per pedigree 2 to 40 affected individuals per pedigree 1880 individuals included in analysis 744 affected (93% genotyped) 744 affected (93% genotyped) 628 unaffected 628 unaffected 508 undetermined phenotype status 508 undetermined phenotype statusGenotyping Subjects were genotyped on 540 florescent dye-labeled microsatellite markers across the genome Subjects were genotyped on 540 florescent dye-labeled microsatellite markers across the genome Genotyping was performed by Myriad genetics Genotyping was performed by Myriad genetics Average spacing of 6 cM between markers Average spacing of 6 cM between markers

17. Methods Phenotype definition Physician confirmed presence or absence of asthma Physician confirmed presence or absence of asthma Based on spirometry measures, medical records and questionnaire Based on spirometry measures, medical records and questionnaire Parametric analyses Mode of inheritance is not well-characterized Mode of inheritance is not well-characterized general dominant and recessive model general dominant and recessive model Disease allele frequency of 0.005 (dom) and 0.05 (rec) Both models assumed penetrance of 50% for disease allele carriers and 0.5% for non-disease carriers

18. Genome-wide results

19. Genome-wide results, cont. A significant 10 result occurred on chromosome 5 LOD = 3.75 LOD = 3.75 ~ 5600:1 odds in favor of linkage ~ 5600:1 odds in favor of linkage Evidence from recessive model Evidence from recessive model Not reported in other genome-wide scans for asthma Not reported in other genome-wide scans for asthma A nearly suggestive result occurred on chromosome 6 LOD = 2.08 LOD = 2.08 ~ 120:1 odds in favor of linkage ~ 120:1 odds in favor of linkage Evidence from dominant model Evidence from dominant model Reported in several other genome-wide scans 11 Reported in several other genome-wide scans 11

20. 1 2 3 4 5 6 7 8 9101112 13 14 15 16 17 18 1920 2122 X Our results in perspective to other published results previously published regions

21. Conclusions Our analysis of extended pedigrees identified a novel asthma susceptibility locus at chromosome 5q21 Our analysis confirmed another region of interest (with nearly suggestive evidence) for an asthma susceptibility locus at 6p21. Inclusion of fine mapping markers in regions of interest will improve localization Future linkage analysis in this resource should address phenotypic heterogeneity of asthma A better understanding of genetic factors for asthma may improve disease outcomes

22. Acknowledgements, 2 People: Alun Thomas Lisa Cannon-Albright Nicola Camp Matt Hegewald Marlene Egger Jim Farnham Steven Backus Institutions: The National Library of Medicine Intermountain Healthcare Myriad Genetics Bayer Pharmaceuticals

23. References 1.American Lung Association, Epidemiology and Statistics Unit, Research and Program Services. Trends in asthma morbidity and mortality. May 2005. 2.Braman SS. The global burden of asthma. Chest. 2006 Jul;130(1 Supp):4S-12S. 3.Wechsler ME, Israel E. The genetics of asthma. Semin Respir Crit Care Med. 2002 Aug;23(4):331- 338. 4.Clark JR, Jenkins MA, Hopper JL, et.al. Evidence for genetic associations between asthma, atopy and bronchial hyperresponsiveness: a study of 8- to 18-year old twins. Am J Respir Crit Care Med. 2000;162(6):2188-2193. 5.Burke W, Fesinmeyer M, Reed K, Hampson L. Family history as a predictor of asthma risk. Am J Prev Med 2003;24:160-169. 6.Hao K, Chen C, Wang B, Yang J, Fang Z, Xu X. Familial aggregation of airway responsiveness: a community-based study. Ann Epidemiol 2005;15:737-743. 7.Teerlink CC, Hegewald M, Cannon-Albright. A genealogical assessment of predisposition to asthma mortality. In press. 8.Ferreira MAR, O'Gorman L, Le Souef P, et al. Robust estimation of experiment-wise P values applied to a genome scan on multiple asthma traits identifies a new region of significant linkage on chromosome 20q13. Am J Hum Genet 2005;77:1075-1085. 9.Contopoulos-Ioannidis DG, Kouri IN, Ioannidis JPA. Genetic predisposition to asthma and atopy. Respiration 2007;74:8-12. 10.Lander E, Kruglyak L. Genetic dissection of complex traits: guidelines for interpreting and reporting linkage results. Nat Genet 1995;11(3):241-247. 11.Nicolae D, Cox NJ, Lester LA, et.al. Fine mapping and positional candidate studies identify HLA-G as an asthma susceptibility gene on chromosome 6p21. Am J Hum Genet. 2005;76:349-357.