Age-related macular degeneration (AMD) is a progressive, blinding disease affecting
millions of elderly individuals worldwide1
2. Several genome-wide association studies (GWAS) have identified common variants
associated with AMD in European-ancestry populations3
4
5
6, and recently, rare genetic variation at CFH, CFI, C3 and C9 were also shown to
strongly associate with AMD in Europeans7
8
9
10. However, there are few such studies in Asians11. Importantly, Asians appear to
have a distinct clinical presentation of the disease (for example, absence of drusen
and minimal fibrous scarring in polypoidal choroidal vasculopathy, a variant of AMD
accounting for 20–55% of Asian patients with exudative AMD) and different responses
to treatment (for example, poorer response to inhibitors of vascular endothelial growth
factor (VEGF) compared with patients of European ancestry)12
13. It remains unclear whether there are differences in underlying genetic characteristics
of AMD between patients of Asian versus European ancestry.
Concurrently, previous GWAS studies provide limited coverage of low-frequency coding
variants, which may result in the loss of function and are often ethnic-specific.
There is thus interest in genetic studies of AMD and other diseases beyond standard-content
GWAS to discover potentially causative coding variants in different ethnic groups.
To address these questions, the Genetics of AMD in Asians Consortium perform a genome-wide
(GWAS) and exome-wide association study (EWAS) of advanced AMD solely on the exudative
(neovascular) disease subtype in East Asians. Compared with standard-content GWAS
arrays, the exome array has significantly increased marker density across the coding
human exome, thus increasing power to detect disease associations located within the
coding frame. EWAS of AMD have not been previously conducted in either Europeans or
Asians. In this paper, we present data from eight independent AMD case–control collections
enrolled across multiple sites in East Asia, totalling 6,345 exudative AMD cases and
15,980 controls. This is the largest sample, to our knowledge, of East Asians ever
assembled for genetic studies of AMD.
Results
Association with previously identified AMD variants
After genotype imputation, synchronization and stringent quality filters were performed,
a total of 4,471,719 SNPs were assessed for the GWAS and 120,027 autosomal coding-frame
SNPs for EWAS from 2,119 AMD cases and 5,691 controls (Table 1). Overall genomic inflation
was very low (λ
gc=1.031; Supplementary Fig. 1), suggesting minimal confounding of the disease association
analysis by population stratification or other systematic study design biases. Data
from the discovery stage analysis confirmed previously identified AMD variants in
ARMS2-HTRA1 rs10490924 (P=1.20 × 10−103), CFH rs10737680 (P=7.54 × 10−38), CETP rs3764261
(P=1.66 × 10−12), ADAMTS9 rs6795735 (P=1.13 × 10−5), C2-CFB rs429608 (P=1.06 × 10−4),
as well as CFI rs4698775 (P=7.5 × 10−4; Supplementary Table 1 and Supplementary Fig.
2). Our data also showed nominal evidence of replication in the same direction as
the initial study for a further three previously reported variants (TGFBR1 rs334353,
APOE rs4420638, and VEGFA rs943080; P<0.05 for each). The remaining 8 out of 17 previously
described SNPs that were non-monomorphic in our East Asian collections did not show
evidence of replication in our study (Supplementary Table 1). A recently described
rare, functional and highly penetrant genetic mutation within CFI (G119R, rs141853578)
shown to confer markedly elevated risk of AMD in Europeans14 was observed to be non-polymorphic
in our East Asian samples (Supplementary Table 1). Similarly, recently described rare
mutations in C3 (K155Q, rs147859257) and C9 (P167S, rs34882957) were also shown to
be non-polymorphic in our East Asian samples (Supplementary Table 1).
Discovery of new SNP variants associated with AMD
Apart from verifying previous observations, our discovery analysis also revealed genome-wide
significant association at C6orf223 (rs2295334 encoding for A231A, P=1.41 × 10−8;
Table 2, Supplementary Table 2 and Supplementary Fig. 2), a novel SNP marker not previously
reported to associate with AMD risk. We observed a further 21 independent SNPs from
distinct loci not previously implicated with susceptibility to AMD showing evidence
of association surpassing P<1 × 10−4. We then brought forward all the 22 markers (Table
2) for replication genotyping in independent sample collections comprising 4,226 exudative
AMD cases and 10,289 controls (Table 1). Replication evidence was compelling for CETP
rs2303790 (encoding D442G; odds ratio (OR)=1.73, P=2.95 × 10−16), as well as for C6orf223
rs2295334 (A231A; OR=0.80, P=5.25 × 10−11), SLC44A4 rs12661281 (D47V; OR=1.22, P=5.13
× 10−6) and FGD6 rs10507047 (Q257R; OR=0.88, P=7.69 × 10−5), leading to genome-wide
significant findings in the meta-analysis of all 6,345 AMD cases and 15,980 controls
(P<5.0 × 10−8 for each of the four loci; Table 2 and Supplementary Table 2). Genotyping
clusters were directly visualized for the top SNPs and confirmed to be of good quality
(Supplementary Fig. 3).
Of note, we did not observe any substantial difference in the association signals
of the most significant SNPs in the subgroup analysis of our AMD cases by typical
neovascular AMD (n=1,083 cases) and polypoidal choroidal vasculopathy (n=1,015 cases;
Supplementary Table 3). The effect size for each of the top SNPs was similar between
the two AMD subgroups.
Conditional analysis
The presence of the mutant CETP 442G (rs2303790) allele is seen only in East Asians
(for example, Chinese, Japanese and Koreans; minor allele frequency (MAF) <5% in our
controls) and not in South Asians, Europeans or Africans. This mutation is independent
from all previously described common, non-coding polymorphisms near the CETP locus
(r
2<0.1; Supplementary Fig. 4). Regional association analysis conditioning on other
known common AMD variants in CETP confirmed the independence of D442G from other nearby
common variants (Supplementary Tables 4 and 5). We also genotyped and assessed multiple,
rare, protein-changing mutations at CETP, including Y74Stop, G331S, N358S and A390P
(Fig. 1). None of them showed association with AMD (Table 3). Mutational load and
haplotypic analysis considering all amino-acid changes within CETP confirmed that
the D442G mutation drove all signals of association between CETP and AMD (Table 4).
C6orf223 is located ~220,000 base pairs downstream of VEGFA and ~150,000 base pairs
from rs943080, a marker previously shown to strongly associate with AMD in Europeans5
6. In this study of AMD in East Asians, the evidence of association for VEGFA rs943080
was only nominally significant (P=0.041 in the discovery stage; Supplementary Table
1). Linkage disequilibrium analysis revealed no correlation between C6orf223 rs2295334
and VEGFA rs943080 (r
2=0.0), with both markers separated by significant recombination events (Fig. 2a).
Logistic regression adjusting for the allele dosage at VEGFA rs943080 did not reveal
any attenuation of the association signal for C6orf223 rs2295334 (P=1.66 × 10−8; Supplementary
Table 6). Regional association analysis including all markers from the genome and
exome array data conditioning on C6orf223 rs2295334 also did not reveal any secondary
signal of association within its 1 Mb flanking region (Supplementary Table 7), thus
pointing to C6orf223 rs2295334 as a novel and uncharacterized genetic risk factor
for exudative AMD in East Asians.
SLC44A4 is located ~116,000 base pairs away from a previously reported AMD locus,
C2-CFB (rs429608)6. Nevertheless, SLC44A4 rs12661281 has no correlation with C2-CFB
rs429608 (r
2=0.01) and showed the strongest evidence of association with AMD within the genomic
region (Fig. 2b), suggesting it to be also a new and uncharacterized risk factor for
AMD. Logistic regression analysis adjusting for allele dosage at C2-CFB rs429608 did
not result in any significant change in magnitude of the association either at SLC44A4
rs12661281, testifying to their mutual independence (ORunconditioned=1.38, P
unconditioned=1.10 × 10−7; ORconditioned for rs429608=1.35, P
conditioned for rs429608=1.49 × 10−6; Supplementary Table 8).
Gene-based tests on mutational load
We next proceeded to conduct gene-based tests on mutational load to further investigate
the role of low-frequency variants in exudative AMD for all the patient collections
in the discovery stage. Gene-based tests are an alternative to single-marker tests
for association, which are often underpowered to detect association with rare variants.
We performed our tests as previously described15. To more directly address the impact
of low-frequency, non-synonymous genetic variants, we considered only 109,296 such
variants with MAF <5%. As a result, we were able to assess a total of 10,736 genes
having at least two such variants using the sequence kernel association optimal (SKAT-O)
test16. We did not detect significant evidence of association (P<5 × 10−8) between
mutational load and AMD at any of the 10,736 genes tested, which are consistent across
all three discovery sample collections. Nonetheless, while looking up on previously
reported 22 genes within 17 distinct loci (Supplementary Table 1) associated with
AMD in European populations, we note nominal evidence of association between genetic
load at CETP (P
unconditioned=5.38 × 10−6), whereby the association was almost entirely driven by
D442G (P
conditioned for D442G=0.96; Table 4) as well as C2 and AMD (P=1.83 × 10−6, Supplementary
Table 9). All the observations exceeding P<1 × 10−4 for gene-based tests on mutational
load summarized across the three discovery collections are appended as Supplementary
Table 9.
CETP 442G, HDL and coronary heart disease
The mutant CETP 442G allele was shown to result in an abnormally functioning CETP
protein17. As CETP is a critical component of the pathways that regulate high-density
lipoprotein cholesterol (HDL-c)18, we assessed this mutation for associations with
serum HDL-c levels using linear regression, with adjustment for age, gender and body
mass index, in three population-based cohorts of Singaporean Chinese19
20
21 and Japanese22 (n=7,102, see details on the study cohorts in Supplementary Methods)
where GWAS data were available. We noted a strong association between 442G allele
and increased HDL-c levels (β=0.174 mmol l−1 per copy of 442G allele; reflecting an
~10% shift within the normal HDL range, P=5.82 × 10−21; Table 5). This effect size
is at least twice that observed for other CETP variants reported in European populations
(Supplementary Table 10)23
24.
Due to its strong effect on serum HDL-c, we assessed whether the mutant CETP 442G
allele conferred any effect on individual susceptibility to coronary heart disease
(CHD) in East Asians. Using 683 CHD cases and 1,281 controls from the Singapore Chinese
Health Study (Supplementary Methods)25, we noted some degree of enrichment of the
HDL-increasing, mutant 442G allele in the controls (2.89%) compared with the cases
(2.42%, OR=0.83), although this did not reach statistical significance (P=0.39). However,
our observations were consistent with a recent study from Japan which analysed CETP
D442G in 4,399 CHD cases and 7,672 controls whereby enrichment of the 442G allele
were also found in the controls (3.4%) compared with the CHD cases (2.8%; OR=0.83,
P=0.02)26. We thus performed a meta-analysis of our study and the Japanese study,
resulting in a consistent protective effect of this mutation with CHD (OR=0.83, P=0.011,
I
2=0.0%; Supplementary Fig. 5).
Discussion
Our studies of exudative AMD in East Asians identified three novel loci (C6orf233,
SLC44A4 and FGD6), two of which (SLC44A4 and FGD6) harbour coding, non-synonymous
variants. These have not been identified in large samples of AMD patients of European
descent6, thus validating the role of searching for coding variations in diverse ethnic
groups to better understand mechanistic basis of complex diseases such as AMD.
Our most interesting findings were the identification of an uncommon East Asian-specific
mutation at CETP (D442G) associated with exudative AMD. The association is the strongest
(per-allele OR=1.70) observed outside of the classical CFH and ARMS2-HTRA1 loci in
East Asians. A common variant (rs3764261) mapping to the intergenic region between
HERPUD1 and CETP was previously linked to AMD in Europeans, yet its effect size was
modest (OR=1.15)5 and independent from the D442G association. The mutant 442G allele
is known to impair CETP function with reduction in plasma CETP mass and activity17
27, and is associated with elevated HDL-c in Japanese families17
27. This allele is absent in European populations28 and appears to be present only
in East Asians, rendering it independent from all other previously described common,
non-coding CETP polymorphisms. We showed that each copy of the dysfunctional 442G
allele confers, on average, a rise in HDL-c levels of 0.174 mmol l−1 and confirmed
findings from the earlier Japanese studies. Given the mean serum HDL-c concentration
is 1.3 mmol l−1, this is a mutation of considerable effect size even from a population-based
perspective. Notably, no other amino-acid substitutions within CETP detected by our
GWAS and EWAS showed any evidence of association with AMD (Fig. 1 and Table 3), suggesting
that the D442G mutation is a possible causative mutation of AMD in East Asians in
the CETP locus. We are not surprised that CETP D442G did not show a clearly significant
protective effect against CHD as increasing lines of evidence more directly implicate
LDL as the driving force for CHD susceptibility29
30.
C6orf223 is a newly mapped gene with yet unknown functional role. Its A231A synonymous
coding change is fivefold rarer in Europeans (MAF=0.03) as compared with Asians (MAF
>0.15, Supplementary Table 2). It is thus unsurprising that the European GWAS efforts
have yet to detect this locus. VEGFA rs943080, a marker strongly associated with AMD
in European-ancestry populations5, is in the vicinity of C6orf223, but its association
with AMD is much weaker in this study of East Asians (Supplementary Tables 1 and 6),
possibly reflecting the differences in therapeutic response to anti-VEGF treatment
between Asians and Europeans13
31.
The D47V mutation within SLC44A4 is not included in most of the routinely used genotyping
arrays, but is now included as part of the exome array used in this study. The genomic
region around the SLC44A4 locus is more complex, being located within the broad MHC
region on Chromosome 6 between HLA class I and class II genes. As this region is very
polymorphic, and allele frequency differences between cases and controls could be
confounded by even minor population stratification, we thus reassessed the associations
with AMD by adjusting for the first 10 principal components (PCs) of genetic ancestry.
We did not observe any change in the association signals observed from our standard
analysis, which adjusted for the first five PCs (analysis adjusting for the top 10
PCs; OR=1.39, P=2.33 × 10−7 in the discovery phase), consistent with previous observations
in Asian studies with well-replicated associations within the MHC region32
33. SLC44A4 encodes for choline transporter protein-4, involved in sodium-dependent
choline uptake by cholinergic neurons34. Defects in SLC44A4 have been linked to sialidosis,
which presents with a spectrum of symptoms including eye abnormalities35.
Prior studies on the basis of European patient collections have identified a total
of four distinct AMD-associated loci on Chromosome 6 alone6. In this light, the burden
of proof for the positive identification of C6orf223 and SLC44A4 (both are also located
on Chromosome 6) is higher than usual due to the need for appropriate considerations
of previously reported variants on the same chromosome. It is reassuring to note that
both C6orf223 and SLC44A1 showed the strongest two signals of association with AMD
outside of CFH, ARMS2-HTRA1 and CETP. Our exhaustive analyses using logistic regression
adjusting for allele dosages at previously described SNP markers suggest that both
C6orf223 and SLC44A1 are unrelated to those previously reported, and thus they likely
represent Asian-specific genetic associations for AMD.
Neither FGD6 nor the genes within its vicinity (Fig. 2c) have ever been previously
implicated in any ocular disorders. FGD6 encodes for FYVE, RhoGEF and PH domain-containing
protein 6, with its functions yet to be characterized. The Q257R mutation is less
than half as common in Europeans (MAF=0.10) compared with East Asians (MAF=0.20–0.30),
again possibly explaining the ability of our study to pick up this genetic effect.
The identified genes were expressed in human retinal pigment epithelium (Supplementary
Table 11). Of the three non-synonymous substitutions, the CETP D442G variant was predicted
by both PolyPhen36 and SIFT37 to likely be causing damage to the protein structure/function,
the FGD6 Q257R variant was predicted only by PolyPhen to be probably damaging but
by SIFT to be tolerated and the SLC44A4 D47V variant was predicted by both tools to
be benign or tolerated. Although the use of both prediction algorithms has been reported
to be moderately sensitive, they suffer from lack of specificity38, and thus more
evidence should be sought with regards to the FDG6 and SLC44A4 non-synonymous variants.
Using HaploReg39, RegulomeDB40 and Encyclopaedia of DNA Elements (ENCODE)41 data,
we identified variants within each of the four LD blocks in the 1000 Genomes Project
(r
2>0.8 and <250 kb from the top SNP) to apply functional annotations relevant to the
regulation of transcription (Supplementary Table 12). In addition to their functions
on amino-acid substitutions, all of the four identified variants lie within a DNase
I hypersensitivity site or in a region where modification of histones is suggestive
of promoter, enhancer and other regulatory activity, and/or have an influence on binding
of transcription factors or effects on a specific regulatory motif. C6orf223 A231A
(rs2295334) and FGD6 Q257R could tag genetic variants that lie in potential transcription-factor
binding sites (Supplementary Table 13). Examination of a recently available large-scale
eQTL mapping database42 indicates that out of the four novel genome-wide significant
SNPs, markers SLC44A4 D47V and FDG6 Q257R could serve as cis-eQTLs. The minor allele
at SLC44A4 D47V (rs12661281) is associated with significantly altered expression of
HSPA1B and CSKN2B, which are located within a 210,000 bp region flanking SLC44A4 D47V.
The minor allele at FDG6 Q257R (rs10507047) is associated with significantly increased
expression of the neighbouring VEZT gene (Supplementary Table 14). Given that these
findings are based on expression in whole blood in European samples, further work
will be needed to elucidate their role in retinal tissue and in Asian samples. Nonetheless,
these could suggest possible alternate mechanisms whereby both non-synonymous substitutions
potentially affect AMD risk apart from directly affecting the protein structure of
their parent genes.
Our study examined mainly the exudative subtype of AMD, and therefore cannot be completely
compared with other studies looking at advanced AMD including the choroidal neovascularization
and geographic atrophy subtypes. We also note substantial differences in inter-ethnic
MAF for most of the previously reported loci associated with AMD in European-ancestry
populations (Supplementary Table 1). This could represent genuine differences in genetic
architecture in AMD between Asians and Europeans, or that the low allele frequency
in either ethnicity could result in insufficient power to replicate genome-wide significant
hits initially observed in either Asians or Europeans. Overall, out of 21 previously
reported SNPs showing strong evidence of association in Europeans, we were able to
replicate 9 of them in our study of East Asians at P<0.05. Taken together with the
three novel loci and one novel variant in CETP in East Asians discovered in this study,
we postulate that the genetic mechanisms of AMD in Asians could, in part, be somewhat
distinct from that in Europeans.
In summary, our genome-wide and exome-wide study of AMD provides new insights into
the genetic mechanisms of AMD in East Asians. Our study highlights the value of searching
for low-frequency, ethnic-specific genetic variants on the coding frame of AMD that
may inform pathogenesis. Although some of the genetic loci conferring disease susceptibility
in East Asians are shared with Europeans (for example, common variation mapping to
CFH, HTRA1 and CETP), we identified significant important differences in the fine-scale
genetic architecture of AMD, which appear specific to East Asians. Such differences
could underpin at least some of the inter-ethnic differences in clinical presentation
and response to specific therapies, including the poorer response to anti-VEGF therapy
in Asians.
Methods
Study design and phenotyping
We performed a GWAS and EWAS on neovascular AMD in East Asians. In the discovery stage,
we included and genotyped 2,119 cases and 5,691 controls from three case–control studies
from Singapore, Hong Kong and Japan. For replication, five independent case–control
studies were conducted in Korea, Japan, and Guangdong, Sichuan and Beijing in China,
totalling 4,226 cases and 10,289 controls. All the studies were performed with the
approval of their local Medical Ethics Committee, and written informed consent was
obtained from all the participants in accordance with the Declaration of Helsinki.
A detailed description of subject recruitment and phenotyping in each sample collection
is provided in Supplementary Methods, and summarized in Table 1. In brief, the diagnosis
of exudative AMD was made at each site by retinal specialists, according to standard
clinical definitions on the basis of detailed ophthalmic examinations, including dilated
fundus photography, fluorescein angiography, indocyanine green angiography and optical
coherence tomography (Table 1). Grading of fluorescein angiograms for the presence
of choroidal neovascularization were performed using a modification from the Macular
Photocoagulation Study43. Indocyanine green angiography was performed to diagnose
definitive polypoidal choroidal vasculopathy, a variant of AMD, using the Japanese
Study Group guidelines44. Cases with other macular diseases such as central serous
chorioretinopathy, high myopia and angioid streaks were excluded. Of the 2,119 exudative
AMD cases included in the discovery phase, 1,083 (51%) were classified as ‘typical
neovascular AMD’, 1,015 (48%) were polypoidal choroidal vasculopathy and 21 (1%) had
one eye with typical neovascular AMD and the other eye with polypoidal choroidal vasculopathy.
At each site, controls subjects without any clinical signs of AMD were either recruited
from eye clinics or enrolled from population-based studies (Supplementary Methods).
Genotyping and imputation
For the discovery stage, GWAS genotyping was performed using the Illumina Human OmniExpress
or Human Hap610-Quad beadchips, and EWAS was done using HumanExome beadchips (Table
1). For replication, genotyping was performed using the MassArray platform (Sequenom),
as well as using Taqman allelic discrimination probes (Applied Biosystems).
Stringent quality control filters were used to remove poorly performing samples and
SNP markers in both the discovery and replication (de-novo genotyping) phases. For
the GWAS, SNPs with a call rate of <95%, MAF of <1%, or showing deviation from Hardy–Weinberg
Equilibrium (P<10−6) were removed from further statistical analysis. For the EWAS,
SNPs with a call rate of <99%, MAF of <0.1% or showing deviation from equilibrium
(P<10−6) were removed. The 99% threshold was used as many SNP markers on the exome
array had MAF <5%, and as such, differential genotyping success rates between cases
and controls as low as 2% could result in false-positive findings. SNPs which were
not monomorphic (whereby at least one heterozygous carrier individual was present)
were included for downstream analysis.
Routine quality control criteria on a per-sample basis were carried out, and poorly
performing samples were removed from further analysis. The remaining samples were
then subjected to biological relationship verification by using the principle of variability
in allele sharing according to the degree of relationship. Identity-by-state information
was derived using the PLINK software45. For those pairs of first-degree relatives
so identified (for example, parent–offspring, full-siblings, as well as monozygous
twins), we removed the sample with the lower call rate before performing PC analysis.
The imputation was carried out using IMPUTE2 version 2.2.2 with ASN population haplotypes
from 1000 Genomes as reference, as described elsewhere46
47
48. Imputed genotypes were called with an impute probability threshold of 0.9 with
all other genotypes classified as missing. Additional quality control filters were
applied to remove SNPs with >1% missingness should the SNP have a MAF <5% in either
cases or controls. For common SNPS with MAF >5%, the filtering criteria were set at
>5% missingness.
Statistical analysis
For the discovery stage, all exudative AMD cases and controls appear well matched
when visualized spatially on PC analysis for each sample collection on a per-country
basis for Hong Kong, Japan and Singapore and according to self-reported ethnicity
(ethnic Chinese for Hong Kong and Singapore, and ethnic Japanese for Japan; Supplementary
Fig. 6), using previously reported criteria49, indicating that population stratification
is unlikely to confound the association results.
For both the discovery and replication stages, analysis of association with exudative
AMD was carried out using 1-degree of freedom score-based tests using logistic regression.
The tests model for a trend-per-copy effect of the minor allele on disease risk. For
the discovery stage, we incorporated the top five PCs of genetic stratification into
the logistic regression model to minimize the effect of residual population stratification50.
We could not adjust for population stratification for the replication stage due to
limited number of SNPs tested. Meta-analysis was conducted using inverse variance
weights for each sample collection, which calculates an overall Z-statistic, corresponding
P value and accompanying per-allele OR for each SNP analysed. Gene-based tests on
mutational load was performed using the SKAT-O test16. The association between CETP
D442G and serum HDL-c level was assessed using linear regression assuming an additive
model of inheritance as previously described23 (due to serum HDL-c being distributed
normally), with adjustment for age, gender and body mass index.
Regional association and PC plots were analysed and plotted using the R statistical
software package.
Power calculations
For the discovery stage (2,119 AMD cases and 5,691 controls), power calculations51
indicated that there is 80% power of detecting loci at P<1 × 10−4 (the threshold of
association for bringing forward SNPs to the replication stage) at MAF as low as 10%
with per-allele OR of 1.30. For rarer variants of higher penetrance, the discovery
stage has 80% power of detecting loci at P<1 × 10−4 at MAF as low as 2% if the per-allele
OR is at least 1.70. The entire sample (6,345 AMD cases and 15,980 controls) has 80%
power to detect loci at P<5.0 × 10−8 at MAF as low as 2% if the per-allele OR is at
least 1.55 or at MAF as low as 9% with per-allele OR of 1.25, in line with the effect
sizes being reported in this study. Supplementary Table 15A shows the power calculations
to detect SNPs at the threshold of P<1 × 10−4 in the discovery stage for bringing
forward to the replication stage. Supplementary Table 15B shows the formal power calculations
in the context of the combined discovery and replication stages.
Author contributions
C.-Y.C., K.Y., L.J.C., J.A., K.-H.P., C.P.P., N.Y., T.Y.W. and C.C.K. designed the
study. C.-Y.C., C.M.G.C., M.M., P.D.C., I.Y.Y., A.L., R.M., A.H.K., S.Y.L., D.W.,
C.M.G.C., B.K.L., Y.S., H.N., Y.A.-K., N.G., A.T., K.M., S.Y., Y.S., H.I., T.I., S.H.,
T.Y.Y.L., H.C., S.T., X.D., F.W., P.Z., B.Z., J.S., J.-M.Y., W.P.K., R.M.v.D., Y.F.,
N.W., G.S.W.T., S.J.P., M.B., L.G., T.N., P.M., P.Z., S.-M.S., M.O., T.M., Y.K., S.J.W.,
H.C., H.-G.Y., J.Y.S., D.H.P., I.T.K., W.C., M.S., S.-J.L., H.W.K., J.E.L., C.K.H.,
T.H.L., S.-K.Y., T.A. and W.T.Y. gathered clinical data. J.P., S.D., I.N., Y.A.-K.,
F.M., P.O.S.T., F.L., X.Z., Y.S., B.G., R.D., Y.L., M.L.H., J.N.F., C.H.W., X.X.,
Jinlong Liang, J.M., X.J., Y.L., Jianjun Liu, K.S., E.N.V., J.X.B., Y.X.Z. and C.C.K.
generated genetic data. C.-Y.C., K.Y., L.J.C., J.A., Lulin Huang, Lvzhen Huang, K.S.S.,
P.C., Jiemin Liao, P.G.O., Y.Y.T. and C.C.K. analysed the data. C.-Y.C., K.Y., L.J.C.,
J.A., Lulin Huang, Lvzhen Huang, E.S.T., X.X.L., Z.Y., K.H.P., C.P.P, N.Y., T.Y.W.
and C.C.K. interpreted the data. C.-Y.C., K.Y., L.J.C., J.A., E.S.T., T.Y.W. and C.C.K.
drafted the paper. All the authors contributed to revision of the paper.
Additional information
How to cite this article: Cheng, C.-Y. et al. New loci and coding variants confer
risk for age-related macular degeneration in East Asians. Nat. Commun. 6:6063 doi:
10.1038/ncomms7063 (2015).
Supplementary Material
Supplementary Information
Supplementary Figures 1-6, Supplementary Tables 1-15, Supplementary Methods and Supplementary
References