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Common Polymorphisms in the SERPINI2 Gene Are Associated with Refractive Error in the 1958 British Birth Cohort

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Common Polymorphisms in the SERPINI2 Gene Are Associated with Refractive Error in the 1958 British Birth Cohort Genetics Common Polymorphisms in the SERPINI2 Gene Are Associated with Refractive Error in the 1958 British Birth Cohort Pirro G. Hysi,1,2,3 Claire L. Simpson,1,3 Yvonne K. Y. Fok,4 Dianne Gerrelli,4 Andrew R. Webster,5 Shomi S. Bhattacharya,5 Christopher J. Hammond,2 Pak C. Sham,6 and Jugnoo S. Rahi1 PURPOSE. To identify heritable genetic factors altering suscep- physiological pathways controlling eye growth and develop- tibility to refractive error in the general population. ment and those controlling glucose metabolism. The findings METHODS. This was a genetic association study of refractive indicate that SERPINI2 is a promising candidate for further error investigating genetic polymorphisms in regions previ- investigations of the genetic susceptibility to myopia. (Invest ously reported through linkage. Two study panels were drawn Ophthalmol Vis Sci. 2012;53:440–447) DOI:10.1167/iovs.10- from the British 1958 Birth Cohort, composed of 2211 persons 5640 44 years of age at the time of visit. Two main outcomes were considered: refractive error as a continuous outcome (spheri- major challenge for the 21st century is to find the genetic cal equivalent) and myopia as a diagnosis (defined as spherical Afactors that influence susceptibility to common complex equivalent equal to or worse thanϪ1.00 diopter). Genotyping diseases and traits. Refractive error is an archetypal complex was initially performed in 1188 subjects from the outer tertiles quantitative trait, with myopia at one end of the distribution of the population distribution, using customized arrays of sin- and hypermetropia at the other. Refractive error is the com- gle nucleotide polymorphisms (SNPs) saturating regions of monest cause of visual impairment globally and has major previously reported highly significant linkage. In a second socioeconomic costs for the affected person and for society as stage, SNPs most significantly associated were validated in a whole; for example North Americans are reported to spend at 1023 more persons. Findings were investigated further least $12 billion annually on glasses, contact lenses, and refrac- 1 through human fetal expression studies. tive surgery. Myopia affects at least 25% of white North Americans and RESULTS. Polymorphisms within the SERPINI2 gene were asso- Europeans2 and up to 60% of Asians but only 5% of Africans.3,4 ciated with refractive error in two different European sub- The prevalence and severity of myopia have been increasing in groups from the 1958 British Birth Cohort (meta-analysis P ϭ recent years, especially in Southeast Asia.3,4 This suggests that, 7.4E-05 for rs9810473). Association was also significant for against a background of genetic susceptibility, changing envi- myopia (best association: OR ϭ 0.80; 95% CI, 0.69–0.93; P ϭ ronmental factors, possibly related to rapid urbanization, play a 0.003 for rs10936538). Expression profiling of SERPINI2 re- role in the development of refractive error. vealed that the gene is expressed in the retina and in other eye Family aggregation studies have shown a correlation be- and CNS tissues. tween parental refractive error and that of their children,5 CONCLUSIONS. The novel association of SERPINI2 with refrac- and twin studies have reported high heritability of 60% to tive error and myopia is suggestive of a possible link between 90% in the general population.6 However, until very re- cently, genetic research has focused on linkage analysis of cohorts of families, often with “syndromic” or “pathologic” From the 1MRC Centre of Epidemiology for Child Health, Institute myopia, whose similarities with common, population-wide of Child Health, University College London, London, United Kingdom; myopia are uncertain. The first linkage study of refractive 2Department of Twin Research and Genetic Epidemiology, Kings Col- error based on dizygotic twins reported four linkage peaks 4 lege London, St. Thomas’ Hospital, London, United Kingdom; Neural on 11p13, 8p23, 3q26, and 4q12.7 We investigated candi- Development Unit, UCL Institute of Child Health, London, United date genes in the top three linkage peaks from that study in Kingdom; 5Institute of Ophthalmology, University College London, London, United Kingdom; and 6Genome Research Center, The Univer- a total of 2211 subjects from the 1958 British Birth Cohort sity of Hong Kong, Hong Kong, SAR, China. and report findings from this candidate gene study and 3These authors contributed equally to the work presented here subsequent expression studies. and should therefore be regarded as equivalent authors. Supported by the Medical Research Council Component Project Grant G0301069 and Health of the Public Project Grant G0000934. METHODS PGH is funded by the EU MyEuropia Marie Curie Research Training Network. Candidate Gene Association Study Submitted for publication April 3, 2010; revised July 13 and The 1958 British Birth Cohort originally consisted of 17,000 persons August 30, 2010, and March 10, July 29, September 30, and October who were born during one week in March 1958 and were subse- 30, 2011; accepted November 1, 2011. quently followed up at intervals8 with physical examination and Disclosure: P.G. Hysi, None; C.L. Simpson, None; Y.K.Y. Fok, collection of data on environmental, social, and lifestyle factors. In None; D. Gerrelli, None; A.R. Webster, None; S.S. Bhattacharya, None; C.J. Hammond, None; P.C. Sham, None; J.S. Rahi, None 2002 to 2003, when the participants were aged 44 to 45 years, a Corresponding author: Jugnoo S. Rahi, MRC Centre of Epidemiol- biomedical survey was undertaken of the 9377 individuals still ogy for Child Health, Institute of Child Health, UCL, 30 Guilford Street, actively participating in the study. Of all the subjects, 2485 (27%) London WC1N 1EH, UK; [email protected]. were selected randomly and underwent noncycloplegic autorefrac- Investigative Ophthalmology & Visual Science, January 2012, Vol. 53, No. 1 440 Copyright 2012 The Association for Research in Vision and Ophthalmology, Inc. Downloaded from jov.arvojournals.org on 09/28/2021 IOVS, January 2012, Vol. 53, No. 1 Polymorphisms in SERPINI2 and Refractive Error 441 tion with an autorefractor (Retinomax 2; Nikon, Tokyo, Japan)9 Ͼ80% calculated from loci genotyped at the first stage was interpreted (equipment costs precluded autorefraction of all subjects). After as proof of identity, and the 644 samples concerned were removed exclusion of 251 subjects for whom no biological material was from the replication panel. The subjects’ Caucasian ancestry in the available and 23 subjects with high anisometropia, this study was replication stage was confirmed by eigenvector analysis.14 The 1023 conducted in two stages. In 1188 subjects in a discovery stage, subjects ultimately included in the replication stage had been geno- fine-mapping was performed of the regions of interest through typed using several platforms, including some of very high density custom genotyping. In a replication stage of another 1023 subjects, (Affymetrix 5 and 6; Illumina Human1M-Duo and Omni 660W-Quad not overlapping with those participating in the first stage, associa- and Illumina HumanHap550 SNP chips). tions were examined in subjects using genomewide data. Autore- fractor measurements in this age group provided a reliable objective Statistical Analysis measure of refractive error at the point at which primary myopia became manifest but before myopia secondary to other ocular After genotyping, SNPs were included in subsequent analytical stages Ͼ disorders occurred.9 This avoided misclassification and ensured if they were in Hardy-Weinberg equilibrium (P 0.01), polymorphic phenotypic stability. Blood was taken for the creation of immortal- (minor allele frequency at least) in the population sample, and above a Ͼ ized cell lines to provide a continuous source of DNA.8 threshold genotyping rate across the sample ( 95%). Refractive error was quantified using the summary measure of We considered two phenotypic outcomes: the spherical equivalent spherical equivalent of refraction (SphE) in diopters (D). We used as transformed using rank-based inverse normal transformation to over- autorefraction readings to calculate SphE for each eye of every subject come nonnormality of phenotype distribution (Fig. 1) and myopia as a in the conventional way (SphE ϭ S ϩ C/2), where S is the sphere and categorical disease status using the criterion of a spherical equivalent of Ϫ1.00 D or more extreme for myopia, with all subjects with0Dor C is the cylinder). Subjects with highly discordant SphE for both eyes 3,15 (within the worst 5% of the sample) were excluded.10,11 Inclusion higher classified as nonmyopes, as in previously published studies. Association was tested using linear and logistic regression models; criteria for the first stage (linkage fine-mapping and SNP association 14 discovery stage) were the mean spherical equivalent within either of PLINK and Stata 10 (Stata Corp, College Station, TX) were used for the analysis. Correction for multiple testing was made either through the outer tertiles of the distribution for this trait (half the participants the Bonferroni method or through phenotype-swapping 1 million in each tertile). The lower tertile spanned the interval Ϫ12.875D to permutations to control for multiple testing where linkage disequilib- Ϫ1.3125D, whereas the upper tertile spanned the interval Ϫ0.3125D rium was so high that the information made available from multiple to ϩ6.50D. As a categorical trait, myopia was determined as a mean SNP loci became highly redundant. spherical equivalent of Ϫ1D or less and nonmyopia as 0D or above. All genotypes obtained at the various stages of the work were Subjects whose spherical equivalent was between 0 and Ϫ1D were not converted with their positive strand alleles as reference. Information included in the qualitative analysis to remove uncertainties of myopia on variation at loci that were not directly genotyped in our assays at classification inherent in any arbitrary dichotomization of quantitative both stages were imputed using the program MACH16 using the traits.12 All participants gave informed written consent to participate in HapMap Phase 2 CEU population as a reference.
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