IMI : Myopia Genetics Report

Special Issue IMI – Myopia Genetics Report Milly S. Tedja, 1,2 Annechien E. G. Haarman, 1,2 Magda A. Meester-Smoor, 1,2 Jaakko Kaprio, 3,4 David A. Mackey, 5–7 Jeremy A. Guggenheim, 8 Christopher J. Hammond, 9 Virginie J. M. Verhoeven, 1,2,10 and Caroline C. W. Klaver 1,2,11 ; for the CREAM Consortium 1 Department of Ophthalmology, Erasmus Medical Center, Rotterdam, the Netherlands 2 Department of Epidemiology, Erasmus Medical Center, Rotterdam, the Netherlands 3 Institute for Molecular Medicine, University of Helsinki, Helsinki, Finland 4 Department of Public Health, University of Helsinki, Helsinki, Finland 5 Centre for Eye Research Australia, Ophthalmology, Department of Surgery, University of Melbourne, Royal Victorian Eye and Ear Hospital, Melbourne, Victoria, Australia 6 Department of Ophthalmology, Menzies Institute of Medical Research, University of Tasmania, Hobart, Tasmania, Australia 7 Centre for Ophthalmology and Visual Science, Lions Eye Institute, University of Western Australia, Perth, Western Australia, Australia 8 School of Optometry and Vision Sciences, Cardiff University, Cardiff, United Kingdom 9 Section of Academic Ophthalmology, School of Life Course Sciences, King’s College London, London, United Kingdom 10 Department of Clinical Genetics, Erasmus Medical Center, Rotterdam, the Netherlands 11 Department of Ophthalmology, Radboud University Medical Center, Nijmegen, the Netherlands Correspondence: Caroline C. W. Klaver, Erasmus Medical Center, Room Na-2808, P.O. Box 2040, 3000 CA, Rotterdam, the Netherlands; [email protected]. MST and AEGH contributed equally to the work presented here and should therefore be regarded as equivalent first authors. VJMV and CCWK contributed equally to the work presented here and should therefore be regarded as equivalent senior authors. See the appendix for the members of the CREAM Consortium. Submitted: October 12, 2018 Accepted: January 9, 2019 Citation: Tedja MS, Haarman AEG, Meester-Smoor MA, et al.; for the CREAM Consortium. IMI – Myopia Genetics Report. Invest Ophthalmol Vis Sci. 2019;60:M89–M105. https:// doi.org/10.1167/iovs.18-25965 The knowledge on the genetic background of refractive error and myopia has expanded dramatically in the past few years. This white paper aims to provide a concise summary of current genetic findings and defines the direction where development is needed. We performed an extensive literature search and conducted informal discussions with key stakeholders. Specific topics reviewed included common refractive error, any and high myopia, and myopia related to syndromes. To date, almost 200 genetic loci have been identified for refractive error and myopia, and risk variants mostly carry low risk but are highly prevalent in the general population. Several genes for secondary syndromic myopia overlap with those for common myopia. Polygenic risk scores show overrepresentation of high myopia in the higher deciles of risk. Annotated genes have a wide variety of functions, and all retinal layers appear to be sites of expression. The current genetic findings offer a world of new molecules involved in myopiagenesis. As the missing heritability is still large, further genetic advances are needed. This Committee recommends expanding large-scale, in-depth genetic studies using complementary big data analytics, consideration of gene-environment effects by thorough measurement of environmental exposures, and focus on subgroups with extreme phenotypes and high familial occurrence. Functional characterization of associated variants is simultaneously needed to bridge the knowledge gap between sequence variance and consequence for eye growth. Keywords: myopia, refractive error, genetics, GWAS, GxE interactions 1. SUMMARY For many years, it has been recognized that myopia is highly heritable, but only recently has significant progress been made in dissecting the genetic background. In particular genome-wide association studies (GWAS) have successfully identified many common genetic variants associated with myopia and refractive error. It is clear that the trait is complex, with many genetic variants of small effect that are expressed in all retinal layers, often with a known function in neurotransmission or extracellular matrix. Exact mechanisms by which these genes function in a retina-to-sclera signaling cascade and other potential pathways remain to be elucidated. The prediction of myopia from genetic risk scores is improving, but whether this knowledge will affect clinical practice is yet unknown. This Committee recommends expanding large-scale genetic studies to further identify the molecular mechanisms through which environmental influences cause myopia (geneby-environment effects), with an ultimate view to develop targeted treatments. 2. KEY POINTS 1. Refractive errors including myopia are caused by a complex interplay between many common genetic factors and environmental factors (near work, outdoor exposure). 2. Early linkage studies and candidate gene studies have identified up to 50 loci and genes, but findings remained mostly unverified in replication studies. Copyright 2019 The Authors iovs.arvojournals.org jISSN: 1552-5783 M89 This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License. Downloaded from iovs.arvojournals.org on 06/12/2019 3. Large consortia performing GWAS enabled identification of common genetic variants associated with refractive error and myopia. 4. The Consortium for Refractive Error and Myopia (CREAM) and 23andMe published findings from GWAS separately, and later combined studies in a GWAS metaanalysis, identifying 161 common variants for refractive error but explaining only approximately 8% of the phenotypic variance of this trait. 5. Polygenic risk scores based on these variants indicate that persons at high genetic risk have an up to 40 times increased risk of myopia compared with persons at low genetic risk. 6. The genetic loci appear to play a role in synaptic transmission, cell-cell adhesion, calcium ion binding, cation channel activity, and the plasma membrane. Many are light-dependent and related to cell-cycle and growth pathways. 7. Pathway analysis confirms the hypothesis for a lightinduced retina-to-sclera signaling pathway for myopia development. 8. Genome-environment-wide interaction studies (GEWIS) assessing variant 3education interaction effects identified nine other loci. Evidence for statistical interaction was also found; those at profound genetic risk with higher education appeared particularly susceptible to developing myopia. 9. As most of the phenotypic variance of refractive errors is still unexplained, larger sample sizes are required with deeper coverage of the genome. 10. The ultimate aim of genetic studies is to discern the molecular signaling cascade and open up new avenues for intervention. 3. INTRODUCTION Although myopia is strongly determined by environmental factors, the trait has long been known to run in families, suggesting a genetic predisposition. The heritability of refractive error, using spherical equivalent as a quantitative trait, has been determined in a number of families and twin studies. 1–8 The estimates resulting from these studies calculated heritabilities from 15% to 98%. 5,7–10 However, it is important to note that this does not necessarily imply that most refractive error is genetic; familial clustering also can be determined by other factors. 11 Like many other traits, common myopia has a complex etiology that is influenced by an interplay of genetic and environmental factors. 12 The current evidence, as summarized in this review, indicates that it is likely to be caused by many genes, each contributing a small effect to the overall myopia risk. The evidence for this has been confirmed by large GWAS. 1–5,7,13,14 Several high, secondary syndromic forms of myopia, such as Marfan, Stickler, and Donnai-Barrow, form the exception, as they inherit predominantly in a Mendelian fashion with one single, highly penetrant, causal gene. 15 This white paper aims to address the recent developments in genetic dissection of common refractive errors, in particular myopia. Up until the era of GWAS, identification of diseaseassociated genes relied on studies using linkage analysis in families or investigating variants in candidate genes. In myopia, these were singularly unsuccessful, and before 2009, there were no genes known for common myopia occurring in the general population. However, with the advent of GWAS, many refractive error genes associated with myopia have been identified, providing potential new insights into the molecular machinery underlying myopia, and perhaps promising leads for future therapies. 4. HERITABILITY Eighty years ago, Sir Duke-Elder was one of the first to recognize a ‘‘ hereditary tendency to myopia.’’ 16 Since then, evidence for familial aggregation has been delivered by various familial clustering, twin, and offspring studies, 1–4 and a genetic predisposition became more widely recognized. Strikingly, the estimates of myopia heritability vary widely among studies, with values as low as 10% 9,10 found in a parent-offspring study in Eskimos, to as high as 98% in a study of female twin pairs 5,7,8 (Table 1). Differences in study design and method of analysis may account for this, but it is also conceivable that the phenotypic variance determined by heritable factors is high in settings in which environmental triggers are limited, and low where they are abundant. Based on literature, heritability of myopia is probably between 60% and 80%. Variation in corneal curvature and axial length contribute to the degree of myopia. 17 Twin studies also estimated a high heritability for most of the individual biometric parameters. 18,19 Correlations between corneal curvature and axial length were at least 64%, 20 suggesting a considerable genetic overlap between the parameters. Studies addressing the inheritance structure of myopia and its endophenotypes identified several models, mostly a combination of additive genetic and environmental effects. 6,18,21,22 Genome-wide complex trait analysis, using high-density genome-wide single-nucleotide polymorphism (SNP) genotype information, was performed in young children from the Avon Longitudinal Study of Parents and Children (ALSPAC), and results suggested that common SNPs explained approximately 35% of the variation in refractive error between unrelated subjects. 23 SNP heritability calculated by linkage disequilibrium score regression in the CREAM Consortium was 21% in European individuals but only 5% in Asian individuals, which could be due to the low representation of this ancestry. 24 In conclusion, the genetic component of myopia and ocular biometry is well recognized, but its magnitude varies in studies depending on the population being studied, the study design, and the methodology. It is important to note that the recent global rise of myopia prevalence is unlikely to be due to genetic factors, but the degree of myopia may still be under genetic control. 25 5. LINKAGE STUDIES A number of linkage studies for myopia were performed in families and high-risk groups before the GWAS era (Fig. 1). 26 Linkage studies have searched for cosegregation of genetic TABLE 1. Heritability Estimates of Refractive Error Subjects Study Heritability Estimate (6SE or 95% CI) Monozygous and dizygous twin pairs Dirani et al. 2006 6 0.88 60.02 (men) (SE) Hammond et al. 2001 21 0.86 (0.83–0.89) Lyhne et al. 2001 7 0.89–0.94 (0.82–0.96) Sibling pair Guggenheim et al. 2007 152 0.90 (0.62–1.12) Peet et al. 2007 153 0.69 (0.58–0.85) Full pedigree Klein et al. 2009 19 0.62 60.13 Parent-offspring pair Lim et al. 2014 154 0.30 (0.27–0.33) IMI – Myopia Genetics Report IOVS jSpecial Issue jVol. 60 jNo. 3 jM90 Downloaded from iovs.arvojournals.org on 06/12/2019 markers (such as cytosine-adenine [CA] repeats) with the trait through pedigrees, and has been successfully applied for many Mendelian disorders. 27 In families with an autosomal dominant inheritance pattern of myopia, this approach helped to identify several independent loci for (high) myopia: MYP 1 to 20, 26,28–30 as well as several other loci. 31–36 Fine-mapping of several of these loci led to candidate genes, such as the IGF1 gene located in the MYP3 locus. 12 Although validation of the same markers failed in these candidate genes, other variants appeared associated with common myopia, suggesting genetic overlap between Mendelian and complex myopia. 37 Linkage studies using a complex inheritance design found five additional loci. 38–42 With the development of new approaches for gene finding, linkage analysis with CA-markers became unfashionable. Nevertheless, segregation and linkage analysis of a variant or region in pedigrees is still a common procedure for finemapping or dissection of disease haplotypes. 6. SECONDARY SYNDROMIC MYOPIA Myopia can accompany other systemic or ocular abnormalities. The secondary syndromic myopias are generally monogenic and have a wide spectrum of clinical presentations. Table 2 summarizes the known syndromic conditions that present with myopia, and Table 3 summarizes the known ocular conditions. 43 Among these disorders are many mental retardation syndromes, such as Angelman (Online Mendelian Inheritance in Man database [OMIM] #105830), Bardet-Biedl (OMIM #209900), and Cohen (OMIM #216550) and Pitt-Hopkins syndrome (OMIM #610954). Myopia also can be a characteristic feature in heritable connective tissue disorders, such as Marfan (OMIM #154700), Stickler (OMIM #108300, #604841, #614134, #614284), and Weill-Marchesani syndrome (OMIM #277600, #608328, #614819, #613195), and several types of Ehlers-Danlos syndrome (OMIM #225400, #601776). A number of inherited retinal dystrophies also present with myopia, most strikingly X-linked retinitis pigmentosa caused by mutations in the RPGR-gene (retinal G protein–coupled receptor) (see Ref. 44 for common gene acronyms) and congenital stationary night blindness. 45 Other eye disorders accompanied by myopia are ocular albinism (OMIM #300500) and Wagner vitreoretinopathy (OMIM #143200). Most genes causing syndromic forms of myopia have not (yet) been implicated in common forms of myopia, except for collagen type II alpha 1 chain (COL2A1) 46,47 and fibrilin 1 (FBN1). 24,48 However, a recent study screened polymorphisms located in and around genes known to cause rare syndromic myopia, and found them to be overrepresented in GWASs on refractive error and myopia. 49 This implies that although rare, pathogenic mutations in these genes have a profound impact on the eye; more benign polymorphisms may have only subtle effects on ocular biometry and refractive error. 7. CANDIDATE GENE STUDIES Candidate genes are generally selected based on their known biological, physiological, or functional relevance to the disease. Although sometimes highly effective, this approach is limited by its reliance on existing knowledge. Another caveat not specific for this approach is that genetic variability across populations can make it difficult to distinguish normal variation from disease-associated variation. 13 In addition, candidate gene studies are very prone to publication bias, and therefore published results are highly selected. Numerous genes have been investigated in candidate gene studies for refractive error traits. Table 4 summarizes all studies that reported statistically significant associations for myopia or ocular refraction. Genes that encode collagens (COL1A1, COL2A1), 46,47 transforming growth factors (TGFb1,TGFb2, TGFb-induced factor homeobox 1 [TGIF1]), 50–52 hepatocyte growth factor and its receptor (HGF,CMET), 53–55 insulin-like FIGURE 1. Historic overview of myopia gene finding. Genes identified using WES are marked as purple. Other loci (linkage studies, GWAS) are marked as red. IMI – Myopia Genetics Report IOVS jSpecial Issue jVol. 60 jNo. 3 jM91 Downloaded from iovs.arvojournals.org on 06/12/2019 growth factor (IGF1), 56,57 matrix metalloproteinases (MMP1, MMP2,MMP3,MMP9,MMP10), 58,59 the lumican gene (LUM), 60 and the ocular developmental gene PAX6, 61 all showed promise in candidate gene studies. Unfortunately, like TABLE 2. Overview of Secondary Syndromic Forms of Myopia: Systemic Syndromes Associated With Myopia Title Gene and Inheritance Pattern Acromelic frontonasal dysostosis ZSWIM6 (AD) Alagille syndrome JAG1 (AD) Alport syndrome COL4A5 (XLD); COL4A3 (AR/ AD) Angelman syndrome UBE3A (IP); CH Bardet-Biedl syndrome ARL6;BBS1;BBS2;BBS4;BBS5; BBS7;BBS9;BBS10;BBS12; CEP290;LZTFL1;MKKS; MKS1;SDCCAG8;TMEM67; TRIM32;TTC8;WDPCP (AR) Beals syndrome FBN2 (AD) Beaulieu-Boycott-Innes syndrome THOC6 (AR) Bohring-Opitz syndrome ASXL1 (AD) Bone fragility and contractures; arterial rupture and deafness PLOD3 (AR) Branchiooculofacial syndrome TFAP2A (AD) Cardiofaciocutaneous syndrome MAP2K2 (AD) Cohen syndrome VPS13B (AR) Cornelia de Lange syndrome NIPBL (AD); HDAC8 (XLD) Cowden syndrome PTEN (AD) Cranioectodermal dysplasia IFT122 (AR) Cutis laxa ATP6V0A2;ALDH18A1 (AR) Danon disease LAMP2 (XLD) Deafness and myopia SLITRK6 (AR) Desanto-Shinawi syndrome WAC (AD) Desbuquois dysplasia CANT1 (AR) Donnai-Barrow syndrome LRP2 (AR) DOORS TBC1D24 (AR) Ehlers-Danlos syndrome COL5A1 (AD); PLOD1 (AR); CHST14 (AR); ADAMTS2 (AR); B3GALT6 (AR); FKBP14 (AR) Emanuel syndrome CH Fibrochondrogenesis COL11A1 (AR) Gyrate atrophy of choroid and retina with/without ornithinemia OAT (AR) Hamamy syndrome IRX5 (AR) Homocystinuria CBS (AR) Joint laxity; short stature; myopia GZF1 (AR) Kaufman oculocerebrofacial syndrome UBE3B (AR) Kenny-Caffey syndrome FAM111A (AD) Kniest dysplasia COL2A1 (AD) Knobloch syndrome COL18A1 (AR) Lamb-Shaffer syndrome SOX5 (AD) Lethal congenital contracture syndrome ERBB3 (AR) Leukodystrophy POLR1C;POLR3A;POLR3B; GJC2 (AR) Linear skin defects with multiple congenital anomalies NDUFB11;COX7B (XLD) Loeys-Dietz syndrome TGFBR1;TGFBR2 (AD) Macrocephaly/megalencephaly syndrome TBC1D7 (AR) Marfan syndrome FBN1 (AD) Marshall syndrome COL11A1 (AD) Microcephaly with/without chorioretinopathy; lymphedema; and/or mental retardation KIF11 (AD) TABLE 2. Continued Title Gene and Inheritance Pattern Mohr-Tranebjaerg syndrome TIMM8A (XLR) Mucolipidosis GNPTAG (AR) Muscular dystrophy TRAPPC11;POMT;POMT1; POMT2;POMGNT1; B3GALNT2;FKRP;DAG1; FKTN (AR) Nephrotic syndrome LAMB2 (AR) Noonan syndrome A2ML1;BRAF;CBL;HRAS; KRAS;MAP2K1;MAP2K2; NRAS;PTPN11;RAF1;RIT1; SOS1;SHOC2;SPRED1 (AD) Oculocutaneous albinism TYR (AR) Oculodentodigital dysplasia GJA1 (AR) Pallister-Killian syndrome CH Papillorenal syndrome PAX2 (AD) Peters-plus syndrome B3GLCT (AR) Pitt-Hopkins syndrome TCF4 (AD) Pontocerebellar hypoplasia CHMP1A (AR) Poretti-Boltshauser syndrome LAMA1 (AR) Prader-Willi syndrome NDN (PC); SNRPN (IP); CH Pseudoxanthoma elasticum ABCC6 (AR) Renal hypomagnesemia CLDN16;CLDN19 (AR) SADDAN FGFR3 (AD) Schaaf-Yang syndrome MAGEL2 (AD) Schimke immunoosseous dysplasia SMARCAL1 (AR) Schuurs-Hoeijmakers syndrome PACS1 (AD) Schwartz-Jampel syndrome HSPG2 (AR) Sengers syndrome AGK (AR) Short stature; hearing loss; retinitis pigmentosa and distinctive facies EXOSC2 (AR) Short stature; optic nerve atrophy; and Pelger-Huet anomaly NBAS (AR) SHORT syndrome PIK3R1 (AD) Short-rib thoracic dysplasia with/without polydactyly WDR19 (AR) Shprintzen-Goldberg syndrome SKI (AD) Singleton-Merten syndrome IFIH1 (AD) Small vessel brain disease with/ without ocular anomalies COL4A1 (AD) Smith-Magenis syndrome RAI1 (AD) Spastic paraplegia HACE1 (AR) Split hand/foot malformation CH Stickler syndrome COL2A1 (AD); COL11A1 (AD); COL9A1 (AR); COL9A2 (AR) Syndromic mental retardation SETD5 (AD); MBD5 (AD); USP9X (XLD); NONO (XLR); RPL10 (XLR); SMS (XLR); ELOVL4 (AR); KDM5C (XLR) Syndromic microphthalmia OTX2;BMP4 (AD) Temtamy syndrome C12orf57 (AR) White-Sutton syndrome POGZ (AD) Zimmermann-Laband syndrome KCNH1 (AD) AD, autosomal dominant; AR, autosomal recessive; CH, chromosomal; IP, imprinting defect; XLD, 3linked dominant; XLR, 3linked recessive. IMI – Myopia Genetics Report IOVS jSpecial Issue jVol. 60 jNo. 3 jM92 Downloaded from iovs.arvojournals.org on 06/12/2019 myopia linkage studies, these studies generally lacked validation by independent studies. 62 Meta-analyses combining data from several candidate gene studies provided evidence for a consistent association between a single SNP in the PAX6 gene and extreme and high myopia. 63 Meta-analyses of the LUM and IGF1 genes did not confirm an association. 64,65 8. GENOME-WIDE ASSOCIATION STUDIES Since the first GWAS in 2005, 66 more than 3000 human GWAS have examined more than 1800 diseases and traits, and thousands of SNP associations have been found. This has greatly augmented our knowledge of human genetics and complex diseases. 14 GWAS genotyping arrays can identify millions of SNPs across the genome in one assay; these variants are generally common and mostly not protein coding. Effect sizes of SNPs associated with disease are mostly small, requiring very large study samples to reach statistical significance. 13,14 Fortunately, technological advances have lowered the costs of genotyping considerably over the years, 67 and GWAS on hundreds of thousands of individuals are becoming more common. 8.1 GWAS of Refractive Errors and Myopia GWAS for myopia have been performed using myopia as a dichotomous outcome or refractive error as a quantitative trait. Several endophenotypes have also been considered: spherical equivalent, axial length, corneal curvature, and age of diagnosis of myopia. Figure 2 provides an overview of all associated loci and nearby genes, their frequency, and effect sizes. 8.1.1 Myopia Case-Control Design. The case-control design using (high) myopia as a dichotomous outcome has been especially popular in East Asia. The first case-control GWAS was performed in a Japanese cohort in 2009. 68 It comprised 830 cases of pathologic myopia (defined as axial length >26 mm) and 1911 controls from the general population. The strongest association was located at 11q24.1, approximately 44 kb upstream of the BH3-like motif containing, cell death inducer (BLID) gene, and conferred odds of higher myopia of 1.37 (95% confidence interval [CI] 1.21– 1.54). Subsequently, a GWAS meta-analysis of two ethnic Chinese cohorts was performed in 287 cases of high myopia (defined as 6 diopters [D]) and 673 controls. The strongest association was for an intronic SNP within the catenin delta 2 (CTNND2) gene on 5p15.2. 69 Neither of these associations met the conventional GWAS threshold (P5310 8 ) for statistical significance due to small sample size. Nevertheless, the locus at 5p15 encompassing the CTNND2 gene was later confirmed by other Asian studies. 70–72 Li et al. 73 studied 102 high myopia cases (defined as 8D with retinopathy) and 335 controls in an ethnic Chinese population. The strongest association (P¼7.70 310 13 ) was a high-frequency variant located in a gene desert within the MYP11 myopia linkage locus on 4q25. In a similar ethnic Han Chinese population of 419 high myopia cases (6 D) and 669 controls, Shi et al. 73,74 identified the strongest association (P¼ 1.91 310 16 ) at an intronic, high-frequency variant within the mitochondrial intermediate peptidase (MIPEP) gene on 13q12. Neither hit has been replicated, even in studies with similar design, phenotypic definition, and ethnic background. In 2013, two papers reported loci for high myopia in Asian populations and these were successfully replicated. Shi et al. 75 studied a Han Chinese population of 665 cases with high myopia (6 D) and 960 controls. Following two-stage replication in three independent cohorts, the most significantly associated variant (P¼8.95 310 14 ) was identified in the vasoactive intestinal peptide receptor 2 (VIPR2) gene within the MYP4 locus, followed by three other variants within a linkage disequilibrium block in the syntrophin beta 1 (SNTB1) gene (P¼1.13 310 8 to 2.13 310 11 ). Khor et al. 76 reported a meta-analysis of four GWAS including 1603 cases of ‘‘ severe’’ myopia and 3427 controls of East Asian ethnicity. After replication and meta-analysis, the SNTB1 gene was confirmed, and a novel variant within the ZFHX1B gene (also known as zinc finger E-box binding homeobox 2 [ZEB2]) reached genome-wide significance (P¼5.79 310 10 ). In 2018, a pathologic myopia case-control study was performed in cohorts of Asian ancestry, using participants with 5.00 D or more myopia with an axial length >26 mm. Fundus photographs were graded pathologic or nonpathologic (N cases ¼828, N controls ¼3624). The researchers found a novel genetic variant in the coiled-coil domain containing 102B (CCDC102B) locus (P¼1.46 310 10 ), which was subsequently replicated in an independent cohort (P¼2.40 310 6 ). This gene is strongly expressed in the RPE and choroid. As myopic TABLE 3. Overview of Secondary Syndromic Forms of Myopia: Ocular Syndromes Associated With Myopia Title Gene and Inheritance Pattern Achromatopsia CNGB3 (AR) Aland Island eye disease GPR143 (XLR) Anterior-segment dysgenesis PITX3 (AD) Bietti crystalline corneoretinal dystrophy CYP4V2 (AD) Blue cone monochromacy OPN1LW;OPN1MW (XLR) Brittle cornea syndrome ZNF469;PRDM5 (AR) Cataract BFSP2;CRYBA2;EPHA2 (AD) Colobomatous macrophthalmia with microcornea CH Cone dystrophy KCNV2 (AD) Cone rod dystrophy C8orf37 (AR); RAB28 (AR); RPGR (XLR); CACNA1F (XLR) Congenital microcoria CH Congenital stationary night blindness NYX (XLR); CACNA1F (XLR); GRM6 (AR); SLC24A1 (AR); LRIT3 (AR); GNB3 (AR); GPR179 (AR) Ectopia lentis et pupillae ADAMTSL4 (AR) High myopia with cataract and vitreoretinal degeneration P3H2 (AR) Keratoconus VSX1 (AD) Leber congenital amaurosis TULP1 (AR) Microcornea, myopic chorioretinal atrophy, and telecanthus ADAMTS18 (AR) Microspherophakia and/or megalocornea, with ectopia lentis and/or secondary glaucoma LTBP2 (AR) Ocular albinism OCA2 (AR) Primary open angle glaucoma MYOC;OPTN (AD) Retinal cone dystrophy KCNV2 (AR) Retinal dystrophy C21orf2 (AR); TUB (AR) Retinitis pigmentosa RP1 (AD); RP2 (XLR); RPGR (XLR); TTC8 (AR) Sveinsson chorioretinal atrophy TEAD1 (AD) Vitreoretinopathy ZNF408 (AD) Wagner vitreoretinopathy VCAN (AD) Weill-Marchesani syndrome ADAMTS10 (AR); FBN1 (AD); LTBP2 (AR); ADAMTS17 (AR) IMI – Myopia Genetics Report IOVS jSpecial Issue jVol. 60 jNo. 3 jM93 Downloaded from iovs.arvojournals.org on 06/12/2019 maculopathy is the primary cause of blindness in high myopia, further functional investigation could be valuable. 77 In Europe, a French case-control GWAS was performed on 192 high myopia cases (6 D) and 1064 controls, and a suggestive association was identified within the MYP10 linkage locus, 3 kb downstream of protein phosphatase 1 regulatory subunit 3B (PPP1R3B). However, this association did not reach genome-wide statistical significance, and no previously reported loci were replicated. 78 Later, in 2016, the direct-toconsumer genetic testing company 23andMe (Mountain View, CA, USA) published a large GWAS on self-reported myopia (N cases ¼106,086 and N controls ¼85,757; all European ancestry), and identified more than 100 novel loci for myopia. 79 Because this study was intended for association analyses between traits, precise locus definitions, post-GWAS quality control, and replication were not performed. 8.1.2 Quantitative Design on Spherical Equivalent. Studies that considered refractive error as a quantitative trait, and included subjects from the general population who displayed the entire range of refractive error, have been more successful. In 2010, the first GWAS for spherical equivalent were carried out in two European populations: a British cohort of 4270 individuals and a Dutch cohort of 5328 individuals. 80,81 Two loci surpassed the GWAS threshold and were replicated: one near the RASGFR1 gene on 15q25.1 (P¼2.70 310 09 ) and the other near GJD2 on 15q14 (P¼2.21 310 14 ). Subsequently, a meta-analysis was performed on 7280 individuals with refractive error from five different cohorts, which included various ethnic populations across different continents, and findings were replicated in 26,953 samples. A novel locus including the RBFOX1 gene on chromosome 16 reached genome-wide significance (P¼3.9 310 9 ). 82 TABLE 4. Summary of Candidate Gene Studies Reporting Positive Association Results With Myopia Gene Study Ethnicity Independent Confirmation Replication in GWAS APLP2 Tkatchenko et al. 2015 131 Caucasian – – BMP2K Liu et al. 2009 155 Chinese – – CHRM1 Lin et al. 2009 156 Han Chinese X 157 – CHRM1 Guggenheim et al. 2010 158 Caucasian X 157 – CMET Khor et al. 2009 55 Chinese – – COL1A1 Inamori et al. 2007 159 Japanese – – COL2A1 Mutti et al. 2007 46 Caucasian – – COL2A1 Metlapally et al. 2009 47 Caucasian – – CRYBA4 Ho et al. 2012 160 Chinese – – HGF Han et al. 2006 54 Han Chinese – – HGF Yanovitch et al. 2009 161 Caucasian – – HGF Veerappan et al. 2010 53 Caucasian – – IGF1 Metlapally et al. 2010 57 Caucasian – – LUM Wang et al. 2006 60 Chinese – – LUM Chen et al. 2009 162 Han Chinese – – LUM Lin et al. 2010 164 Chinese – – LUM Guggenheim et al. 2010 158 Caucasian – – MFN1 Andrew et al. 2008 164 Caucasian X 165 – MMP1 Wojciechowski et al. 2010 130 Amish – – MMP1 Wojciechowski et al. 2013 59 Caucasian – – MMP10 Wojciechowski et al. 2013 59 Caucasian – – MMP2 Wojciechowski et al. 2010 130 Amish – – MMP2 Wojciechowski et al. 2013 59 Caucasian – – MMP3 Hall et al. 2009 58 Caucasian – – MMP9 Hall et al. 2009 58 Caucasian – – MYOC Tang et al. 2007 63 Chinese – – MYOC Vatavuk et al. 2009 167 Caucasian – – MYOC Zayats et al. 2009 168 Caucasian – – PAX6 Tsai et al. 2008 169 Chinese – – PAX6 Ng et al. 2009 170 Han Chinese – – PAX6 Han et al. 2009 171 Han Chinese – – PAX6 Miyake et al. 2012 172 Japanese – – PAX6 Kanemaki et al. 2015 173 Japanese – – PSARL Andrew et al. 2008 164 Caucasian – – SOX2T Andrew et al. 2008 164 Caucasian – TGFb1Lin et al. 2006 50 Chinese – X 24 TGFb1Zha et al. 2009 174 Chinese – X 24 TGFb1Khor et al. 2010 56 Chinese – X 24 TGFb1Rasool et al. 2013 175 Indian – X 24 TGFb2Lin et al. 2009 51 Han Chinese – – TGIF Lam et al. 2003 52 Chinese – – TGIF1 Ahmed et al. 2014 52,176 Indian – – LAMA1 Zhao et al. 2011 177 Chinese – – UMODL1 Nishizaki et al. 2009 178 Japanese – – X indicates independent conformation or replication in GWAS study with reference included. IMI – Myopia Genetics Report IOVS jSpecial Issue jVol. 60 jNo. 3 jM94 Downloaded from iovs.arvojournals.org on 06/12/2019 These collaborations paved the way for the formation of a large consortium to achieve higher statistical power for gene finding. CREAM was established in 2010 and included researchers and cohorts from the United States, Europe, Asia, and Australia. Its first collaborative work was replication of SNPs in the previously identified 15q14 loci. 83 Other studies followed this approach, and confirmed 15q14 as well as the 15q25 locus. 84,85 Subsequently, CREAM conducted a GWAS meta-analysis based on HapMapII imputation 86 with 35 participating studies comprising 37,382 individuals of European descent and 12,332 of Southeast Asian ancestry with data on GWAS and spherical equivalent. This study enabled replication of GJD2,RASGRF1, and RFBOX1 and identification of 23 novel loci at genome-wide significance: BICC1,BMP2,CACNA1D, CD55,CHD7,CHRNG,CYP26A1,GRIA4,KCNJ2,KCNQ5, LOC100506035,LAMA2,MYO1D,PCCA,TJP2,PTPRR, SHISA6,PRSS56,RDH5,RORB,SIX6,TOX, and ZMAT472. 87 Meanwhile, 23andMe performed a contemporaneous large GWAS on 55,177 individuals of European descent by using a survival analysis, based on the first release of 1000G 88 (a catalog of human genetic variation). Its analysis was based on self-reported presence of myopia and age of spectacle wear as a proxy for severity. 23andMe also replicated GJD2,RASGRF1, and RFBOX1 and identified 11 new loci: BMP3,BMP4,DLG2, DLX1,KCNMA1,LRRC4C,PABPCP2,PDE11A,RGR,ZBTB38, ZIC2. 89 Of the 22 loci discovered by CREAM, 8 were replicated by 23andMe, and 16 of the 20 loci identified by 23andMe were confirmed by CREAM. This was surprising, as the studies used very different phenotyping methods. In addition, the effect sizes of 25 loci were very similar, despite analyses on different scales: diopters for CREAM and hazard ratios for 23andMe. 90 After these two publications, replication studies provided validation for KCNQ5,GJD2,RASGRF1,BICC1,CD55, CYP26A1,LRRC4C,LAMA2,PRSS56,RFBOX1,TOX,ZIC2, ZMAT4, and B4GALNT2 in per-SNP analyses, and for GRIA4, BMP2,BMP4,SFRP1,SH3GL2, and EHBP1L1 in gene-based analyses. 91–96 Although CREAM and 23andMe found a large number of loci, only approximately 3% of the phenotypic variance of refractive error was explained. 87,89 Larger GWAS meta-analyses were clearly needed, and the two large studies combined efforts. This new GWAS meta-analysis was based on the phase 1 version 3 release of 1000G, included 160,420 participants, and findings were replicated in the UK Biobank (95,505 participants). Using this approach, the number of validated refractive error loci increased to 161. A high genetic correlation between European and Asian individuals (>0.78) was found, implying that the genetic architecture of refractive error is quite similar for European and Asian individuals. Taken together, these genetic variants accounted for 7.8% of the explained phenotypic variance, leaving room for improvement. Even so, polygenic risk scores, which are constructed by the sum of effect sizes of all risk variants per individual depending on their genotypes, were well able to distinguish individuals with hyperopia from those with myopia at the lower and higher deciles. Interestingly, those in the highest decile had a 40-fold greater risk of myopia. The predictive value (area under the curve) of these risk scores for myopia versus hyperopia, adjusted for age and sex, was 0.77 (95% CI 0.75–0.79). The next step will include GWAS on even larger sample sizes. Although this will improve the explained phenotypic variance, it is unlikely that GWAS will uncover the entire FIGURE 2. Effect sizes of common and rare variants for myopia and refractive error. Overview of SNPs and annotated genes found in the most recent GWAS meta-analysis. 24 The x-axis displays the minor allele frequency of each SNP; y-axis displays the effect size of the individual SNP in diopters; We transformed the z-scores of the fixed effect meta-analysis between CREAM (refractive error) and 23andMe (age of diagnosis of myopia) into effect sizes in diopters with the following formula 24 :SE ¼ffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi 1 2NMAFð1MAFÞ q. IMI – Myopia Genetics Report IOVS jSpecial Issue jVol. 60 jNo. 3 jM95 Downloaded from iovs.arvojournals.org on 06/12/2019 missing heritability. SNP arrays do not include rare variants, nor do they address gene-environment and gene-gene interactions, or epigenetic effects. 8.1.3 GWAS on Refractive Error Endophenotypes. As myopia is mostly due to increased axial length, researchers have used this parameter as a myopia proxy or ‘‘ endophenotype.’’ The first axial length GWAS examined 4944 individuals of East and Southeast Asian ancestry, and a locus on 1q41 containing the zinc finger pseudogene ZC3H11B reached genome-wide significance (P¼4.38 310 10 ). 82,97 A much larger GWAS meta-analysis of axial length comprised 12,531 European individuals and 8216 Asian individuals. 93 This study identified eight novel genome-wide significant loci (RSPO1, C3orf26,LAMA2,GJD2,ZNRF3,CD55,MIP,ALPPL2), and also replicated the ZC3H11B gene. Notably, five of these loci had been associated with refractive error in previous GWAS. Several relatively small GWAS have been performed for corneal curvature, and identified associations with FRAP1, PDGFRA (also associated with eye size), CMPK1,and RBP3. 93,98–101 More recently Miyake et al. 101,102 published a two-stage GWAS for three myopia-related traits: axial length, corneal curvature, and refractive error. The study was performed in 9804 Japanese individuals, with trans-ethnic replication in Chinese and Caucasian individuals. A novel gene, WNT7B, was identified for axial length (P¼3.9 310 13 ) and corneal curvature (P¼2.9 310 40 ), and the previously reported association with GJD2 and refractive error was replicated. 8.2 Genome-Wide Pathway Analyses The main goal of GWAS is to improve insight on the molecules involved in disease, and help identify disease mechanisms. For myopia, a retina-to-sclera signaling cascade had been proposed for many years (see accompanying paper IMI – Report on Experimental Models of Emmetropization and Myopia 103 ), but knowledge on its molecular drivers was limited. Several attempts were made to translate the findings from refractive error GWAS into this cascade. 87,89,104 Here we provide an overview of genes annotated to the risk variants and their relationship to the underlying biological mechanism. Deducted from the CREAM GWAS, pathways included neurotransmission (GRIA4), ion transport (KCNQ5), retinoic acid metabolism (RDH5), extracellular matrix remodeling (LAMA2,BMP2), and eye development (SIX6,PRSS56). Likewise, 23andMe proposed extracellular matrix remodeling (LAMA2,ANTXR2), the visual cycle (RDH5, RGR,KCNQ5), neuronal development (KCNMA1,RBFOX1,LRRC4C,NGL-1, DLG2,TJP2), eye and body growth (PRSS56,BMP4,ZBTB38, DLX1), and retinal ganglion cells (ZIC2,SFRP1) 105 as functions. Hysi et al. 106 performed pathway analyses using both the CREAM and 23andMe GWAS, and reported that plasma membrane, cell-cell adhesion, synaptic transmission, calcium ion binding, and cation channel activity were significantly overrepresented in refractive error in two British cohorts. Furthermore, by examining known protein-protein interactions, the investigators identified that many genes are related to cell-cycle and growth pathways, such as the MAPK and TGF-beta/SMAD pathways. The latest update on pathway analysis in myopia stems from the meta-GWAS from CREAM and 23andMe. 24 TGF-beta signaling pathway was a key player; the association with the DRD1 gene provided genetic evidence for a dopamine pathway. Most genes were known to play a role in the eye, 107 and most significant gene sets were ‘‘ abnormal photoreceptor inner segment morphology’’ (Mammalian Phenotype Ontology [MP] 0003730; P¼1.79 310 7 ), ‘‘thin retinal outer nuclear layer’’ (MP 0008515), ‘‘ detection of light stimulus’’ (Gene Ontology [GO] 0009583), ‘‘ nonmotile primary cilium’’ (GO 0031513), and ‘‘ abnormal anterior-eye-segment morphology’’ (MP 0005193). Notably, RGR,RP1L1,RORB, and GNB3 were present in all of these meta-gene sets. Taken together, retinal cell physiology and light processing are clearly prominent mechanisms for refractive error development, and all cell types of the neurosensory retina, RPE, vascular endothelium, and extracellular matrix appear to be involved (Fig. 3). Novel mechanisms included rod-and-cone bipolar synaptic neurotransmission, anterior-segment morphology, and angiogenesis. 24 9. WHOLE-EXOME AND WHOLE-GENOME SEQUENCING Unlike GWAS, whole-exome sequencing (WES) and wholegenome sequencing (WGS) have the potential to investigate rare variants. Exomes are interesting, as they directly contribute to protein translation, but they constitute only approximately 1% of the entire genome. WGS allows for identification of variants across the entire genome, but requires a highthroughput computational infrastructure and remains costly. WES has been conducted primarily in case-control studies of early-onset high myopia or in specific families with a particular phenotype (i.e., myopic anisometropia) or inheritance pattern (i.e., X-linked). 108–111 Several novel mutations in known myopia genes were identified this way: CCDC111, 109 NDUFAF7, 110 P4HA2, 108 SCO2, 112 UNC5D, 111 BSG, 113 ARR3, 114 LOXL3, 115 SLC39A5, 116 LRPAP1, 117 CTSH, 117 ZNF644. 118,119 Although most genetic variants displayed an autosomal dominant hereditary pattern, 108,112,118,119 X-linked heterozygous mutations were identified in ARR3, only in female family members. 114 The functions of these novel genes include DNA transcription (CCDC111,ZNF644), mitochondrial function (NDUFAF7,SCO2), collagen synthesis (P4HA2), cell signaling (UNC5D,BSG), retina-specific signal transduction (ARR3), TGF-beta pathway (LOXL3,SLC39A5,LRPAP1), and degradation of proteins in lysosomes (CTSH). Jiang et al. 119 investigated family members with high myopia and identified new mutations in LDL receptor related protein associated protein 1 (LRPAP1), cathepsin H (CTSH ), zinc finger protein 644 isoform 1 (ZNF644), solute carrier family 39 (metal ion transporter) member 5 (SLC39A5), and SCO2, cytochrome c oxidase assembly protein (SCO2). 119 Many clinicians have noticed that retinal dystrophies and ocular developmental disorders often coincide with myopia. 115 This triggered Sun et al. 120 to evaluate variants in a large number of retinal dystrophy genes in early-onset high myopia in 298 unrelated myopia probands and their families, and they thereby identified 29 potentially pathogenic mutations in COL2A1,COL11A1,PRPH2,FBN1,GNAT1,OPA1,PAX2, GUCY2D,TSPAN12,CACNA1F, and RPGR, and most had an autosomal dominant inheritance pattern. Kloss et al. 121 performed WES in 14 families with high myopia, and identified 104 new genetic variants located in both known MYP loci (e.g., AGRN,EME1, and HOXA2) and in new loci (e.g., ATL3 and AKAP12). To date, WGS has not been conducted for myopia or refractive error, most likely due to the reasons mentioned above. When costs for WGS decrease, these studies will undoubtedly be conceived. 10. GENE-ENVIRONMENT INTERACTION It has become clear that environmental factors are driving the recent epidemic rise in the prevalence of myopia. 122–126 To date, the most influential and consistent environmental factor is education. Studies have estimated that individuals going onto IMI – Myopia Genetics Report IOVS jSpecial Issue jVol. 60 jNo. 3 jM96 Downloaded from iovs.arvojournals.org on 06/12/2019 higher education have double the myopia prevalence comparedwiththosewholeaveschoolafteronlyprimary education. 127–129 Education has been a primary focus for gene-environment (GxE) interaction analyses in myopia. GxE studies have the potential to show modification of the effect of risk variants by environmental exposures, but can also reveal genetic associations that were hidden in unexposed individuals. One of the first GxE studies for myopia investigated variants in matrix metalloproteinases genes (MMP1–MMP10). Two SNPs (rs1939008 and rs9928731) that were first found to be associated with refraction in Amish families were also associated in a lower but not in the higher education group of the Age-Related Eye Disease Study (AREDS) study. These results suggest that variants in these genes may play a role in refractive variation in individuals not exposed to myopic triggers. 59,130 In contrast, a study combining human GWAS data and animal models of myopia provided an experimental example of GxE interaction involving a rare variant in the APLP2-gene only in children exposed to large amounts of daily reading. 131 In addition, an analysis performed in five Singapore cohorts found risk variants in DNAH9,GJD2, and ZMAT4 that were more strongly associated in individuals who achieved higher secondary or university education. 132 Significant biological interaction between education and other risk variants was studied using a genetic risk score of all known risk variants at the time (n¼26) derived from the CREAM metaGWAS. 133 European subjects with a high genetic load in combination with university-level education had a far greater risk of myopia than those with only one of these two factors. A study investigating GxE interactions in children and the major environmental risk-factors, nearwork, time outdoors, and 39 SNPs derived from the CREAM meta-GWAS revealed nominal evidence of interaction with nearwork (top variant in ZMAT4). 133,134 GEWIS, using all variants from the CREAM meta-GWAS, revealed three novel loci (AREG,GABRR1, and PDE10A)for GxE in Asian populations, whereas no interaction effects were observed in Europeans due to many reasons, such as the quantitative differences in the intensity of nearwork during childhood. 48 Up to now, there is no robust evidence that there are fundamental differences in the genetic background of myopia risk between European and Asian individuals. 11. MENDELIAN RANDOMIZATION Mendelian randomization (MR) is a method that allows one to test or estimate a causal effect from observational data in the presence of confounding factors. MR is a specific type of instrumental variable analysis that uses genetic variants with well-understood effects on exposures or modifiable biomarkers. 135,136 Importantly, the SNP must affect the disease status only indirectly via its effect on the exposure of interest. 137 MR is particularly valuable in situations in which randomized controlled trials are not feasible, where it is applied to help elucidate biological pathways. Currently, three studies have been published on MR in refractive error and myopia. The first, published in 2016, explored the effect of education on myopia. 138 This study constructed polygenic risk scores of genetic variants found in GWAS for educational attainment, and used these as the instrumental variable. Subsequently, results of three cohorts (Cooperative Health Research in the Region Augsburg [KORA], FIGURE 3. Schematic overview of expression in retinal cells of refractive error and syndromic myopia genes according to literature. Bold: genes identified for both common refractive error and in syndromic myopia. IMI – Myopia Genetics Report IOVS jSpecial Issue jVol. 60 jNo. 3 jM97 Downloaded from iovs.arvojournals.org on 06/12/2019 9 Department of Ophthalmology and Visual Sciences, The Chinese University of Hong Kong, Hong Kong Eye Hospital, Kowloon, Hong Kong 10 Department of Ophthalmology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 11 Ocular Epidemiology Research Group, Singapore Eye Research Institute, Singapore National Eye Centre, Singapore 12 Ophthalmology & Visual Sciences Academic Clinical Program (Eye ACP), DUKE-NUS Medical School, Singapore 13 Division of Epidemiology and Clinical Applications, National Eye Institute/National Institutes of Health, Bethesda, Maryland, United States 14 Department of Ophthalmology, Flinders University, Adelaide, Australia 15 Great Ormond Street Institute of Child Health, University College London, London, United Kingdom 16 Department of Ophthalmology and Visual Sciences, John Moran Eye Center, University of Utah, Salt Lake City, Utah, United States 17 Universit´ e de Bordeaux, INSERM, Bordeaux Population Health Research Center, Team LEHA, UMR 1219, F-33000 Bordeaux, France 18 State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China 19 Department of Epidemiology, Erasmus Medical Center, Rotterdam, The Netherlands 20 Translational Research Institute, University of Queensland Diamantina Institute, Brisbane, Queensland, Australia 21 MRC Integrative Epidemiology Unit, University of Bristol, Bristol, United Kingdom 22 Department of Population Health Sciences, Bristol Medical School, Bristol, United Kingdom 23 Centre for Quantitative Medicine, DUKE-National University of Singapore, Singapore 24 University Hospital ‘San Giovanni di Dio,’ Cagliari, Italy 25 NIHR Biomedical Research Centre, Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, London, United Kingdom 26 Statistical Genetics, QIMR Berghofer Medical Research Institute, Brisbane, Australia 27 Department of Clinical Genetics, Erasmus Medical Center, Rotterdam, the Netherlands 28 Department of Ophthalmology, Erasmus Medical Center, Rotterdam, the Netherlands 29 School of Optometry & Vision Sciences, Cardiff University, Cardiff, United Kingdom 30 Department of Statistical Science, School of Mathematics, Sun Yat-Sen University, Guangzhou, China 31 Institute of Genomics, University of Tartu, Tartu, Estonia 32 Section of Academic Ophthalmology, School of Life Course Sciences, King’s College London, London, United Kingdom 33 MRC Human Genetics Unit, MRC Institute of Genetics & Molecular Medicine, University of Edinburgh, Edinburgh, United Kingdom 34 Centre for Ophthalmology and Visual Science, Lions Eye Institute, University of Western Australia, Perth, Australia 35 Department of Ophthalmology, Columbia University, New York, United States 36 Department of Population and Quantitative Health Sciences, Case Western Reserve University, Cleveland, Ohio, United States 37 Department of Ophthalmology and Visual Sciences, Case Western Reserve University and University Hospitals Eye Institute, Cleveland, Ohio, United States 38 Department of Genetics, Case Western Reserve University, Cleveland, Ohio, United States 39 Department of Ophthalmology, Medical Faculty Mannheim of the Ruprecht-Karls-University of Heidelberg, Mannheim, Germany 40 Beijing Tongren Eye Center, Beijing Tongren Hospital, Beijing Institute of Ophthalmology, Beijing Key Laboratory of Ophthalmology and Visual Sciences, Capital Medical University, Beijing, China 41 Department of Clinical Physiology, Tampere University Hospital and School of Medicine, University of Tampere, Tampere, Finland 42 Finnish Cardiovascular Research Center, Faculty of Medicine and Life Sciences, University of Tampere, Tampere, Finland 43 Department of Public Health, University of Helsinki, Helsinki, Finland 44 Institute for Molecular Medicine Finland FIMM, HiLIFE Unit, University of Helsinki, Helsinki, Finland 45 Department of Public Health and Primary Care, University of Cambridge, Cambridge, United Kingdom 46 Department of Ophthalmology, Radboud University Medical Center, Nijmegen, the Netherlands 47 Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, Wisconsin, United States 48 Department of Clinical Chemistry, Finnish Cardiovascular Research Center-Tampere, Faculty of Medicine and Life Sciences, University of Tampere, Tampere, Finland 49 Department of Clinical Chemistry, Fimlab Laboratories, University of Tampere, Tampere, Finland 50 National Human Genome Research Institute, National Institutes of Health, Baltimore, Maryland, United States 51 Genetic Epidemiology, QIMR Berghofer Medical Research Institute, Brisbane, Australia 52 Department of Ophthalmology, Yokohama City University School of Medicine, Yokohama, Kanagawa, Japan 53 Department of Ophthalmology and Visual Sciences, Kyoto University Graduate School of Medicine, Kyoto, Japan 54 Department of Ophthalmology, University Medical Center of the Johannes Gutenberg University Mainz, Mainz, Germany 55 Institute of Neurogenomics, Helmholtz Zentrum M¨ unchen, German Research Centre for Environmental Health, Neuherberg, Germany 56 Department of Ophthalmology, Central Hospital of Central Finland, Jyv¨ askyl¨a, Finland 57 Gerontology Research Center, Faculty of Sport and Health Sciences, University of Jyv¨askyl¨a, Jyv¨askyl¨a, Finland 58 Program in Genetics and Genome Biology, Hospital for Sick Children and University of Toronto, Toronto, Ontario, Canada 59 Gen-info Ltd, Zagreb, Croatia 60 University of Split School of Medicine, Soltanska 2, Split, Croatia 61 Ulverscroft Vision Research Group, University College London, London, United Kingdom 62 Research Centre of Applied and Preventive Cardiovascular Medicine, University of Turku, Turku, Finland 63 Department of Clinical Physiology and Nuclear Medicine, Turku University Hospital, Turku, Finland 64 Myopia Research Group, Singapore Eye Research Institute, Singapore National Eye Centre, Singapore 65 Saw Swee Hock School of Public Health, National University Health Systems, National University of Singapore, Singapore 66 Department of Ophthalmology, University of Pennsylvania, Philadelphia, Pennsylvania, United States 67 Department of Genetics, Genomics and Informatics, University of Tennessee Health Sciences Center, Memphis, Tennessee, United States IMI – Myopia Genetics Report IOVS jSpecial Issue jVol. 60 jNo. 3 jM104 Downloaded from iovs.arvojournals.org on 06/12/2019 68 Department of Ophthalmology, University of Helsinki and Helsinki University Hospital, Helsinki, Finland 69 Beijing Tongren Eye Center, Beijing Key Laboratory of Intraocular Tumor Diagnosis and Treatment, Beijing Ophthalmology & Visual Sciences Key Lab, Beijing Tongren Hospital, Capital Medical University, Beijing, China 70 Department of Epidemiology and Medicine, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, United States 71 Wilmer Eye Institute, Johns Hopkins Medical Institutions, Baltimore, Maryland, United States 72 Department of Ophthalmology, Otsu Red Cross Hospital, Nagara, Japan 73 Centre for Myopia Research, School of Optometry, The Hong Kong Polytechnic University, Hong Kong 74 Department of Health Technology and Informatics, The Hong Kong Polytechnic University, Hong Kong 75 School of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, China IMI – Myopia Genetics Report IOVS jSpecial Issue jVol. 60 jNo. 3 jM105 Downloaded from iovs.arvojournals.org on 06/12/2019