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2007 Nature Publishing Group http://www.nature.com/naturegenetics

A common CFH haplotype, with deletion of CFHR1 and CFHR3, is associated with lower risk of age-related macular degeneration
Anne E Hughes1, Nick Orr1, Hossein Esfandiary1, Martha Diaz-Torres2, Timothy Goodship2 & Usha Chakravarthy3
Age-related macular degeneration (AMD; OMIM #603075) is the most frequent cause of visual impairment in the elderly population, with severe disease affecting nearly 10% of individuals of European descent over the age of 75 years. It is a complex disease in which genetic and environmental factors contribute to susceptibility. Complement factor H (CFH) has recently been identied as a major AMD susceptibility gene, and the Y402H polymorphism has been proposed as the likely causative factor. We genotyped polymorphisms spanning the cluster of CFH and ve CFHrelated genes on chromosome 1q23 in 173 individuals with severe neovascular AMD and 170 elderly controls with no signs of AMD. Detailed analysis showed a common haplotype associated with decreased risk of AMD that was present on 20% of chromosomes of controls and 8% of chromosomes of individuals with AMD. We found that this haplotype carried a deletion of CFHR1 and CFHR3, and the proteins encoded by these genes were absent in serum of homozygotes. The protective effect of the deletion haplotype cannot be attributed to linkage disequilibrium with Y402H and was replicated in an independent sample. CFH and the closely related genes CFHR3, CFHR1, CFHR4, CFHR2 and CFHR5 are arranged in tandem on chromosome 1q23, where they span 355 kb at the proximal end of a cluster of genes involved in regulation of complement activation. They share an extremely high similarity, which reaches 98% between the 3 exons of CFH and CFHR1. The complement cascade is implicated in formation of drusen, deposits that form between Bruchs membrane and the retinal pigment epithelium in eyes showing early signs of AMD1. The participants in our experiments were recruited from ophthalmology clinics and had choroidal neovascularization associated with the more severe exudative or wet form of AMD. Our age-matched controls had no signs of age-related macular disease. We typed 30 SNPs spanning the CFH region selected to ascertain maximum haplotype information for each of the genes. Our data conrmed the strong association between CFH and AMD25, which was evident by assessing SNPs both individually (Table 1) and by haplotype (Table 2). There was extensive linkage disequilibrium (LD) throughout the region. With the exception of rs512900 at the start of intron 1 of CFH, rs2019727 (CFH intron 10), rs1065489 (CFH exon 19) and rs10922152 (CFHR5 intron 3), all SNPs typed in any of the genes were placed within four large haplotype blocks of 36, 27, 84 and 60 kb, respectively (Fig. 1 and Supplementary Fig. 1 online). LD disintegrated sharply before F13B, 32 kb distal of CFHR5 (data not shown). Haplotypes ranged in effect from strongly detrimental to highly protective of AMD, with those in blocks 2, 3 and 4 being marginally more strongly associated with disease. In block 1, rs1061170 (Y402H) characterized the haplotype associated with most increased risk of AMD, conferring an odds ratio (OR) of 1.99. Two protective haplotypes were evident. The more strongly protective haplotype (haplotype 5, marked in Fig. 1b) showed a solid spine of LD across blocks 2 and 3 and was found in nearly 20% of chromosomes of controls and 7.8% of AMD chromosomes, with an OR of 0.35 (inverse OR 2.9). Logistic regression analysis conrmed that variation at Y402H and haplotype 5 each conferred a signicant independent effect on AMD status of similar magnitude (Supplementary Table 1 online). We conrmed the effect of Y402H and of block 2 haplotypes by typing rs1061170 and four tagging SNPs for block 2 (rs2274700, rs3753396, rs419137 and rs2284664) in a second group of 192 individuals with end-stage AMD and 192 elderly controls who had not undergone eye examination (Supplementary Table 2 online). The highly protective haplotype 5 was found in 8.7% of chromosomes in this AMD group and in 15.9% of controls, up to 10% of whom may be expected to have AMD. Meta-analysis combining data from all our individuals with AMD and our negative controls with published data from ref. 5 showed an overall OR of 0.4 (95% condence interval (c.i.) 0.30.5). Similarly, meta-analysis of Y402H, which also included data from ref. 6, showed an OR of 2.5 (95% c.i. 2.22.8; Supplementary Note online).

1Department of Medical Genetics, Queens University, Belfast, Belfast BT12 6BL, UK. 2Institute of Human Genetics, University of Newcastle upon Tyne, Newcastle upon Tyne NE1 3BZ, UK. 3Centre for Vision Science and Vascular Biology, Queens University, Belfast, Belfast BT12 6BL, UK. Correspondence should be addressed to A.H. (A.Hughes@qub.ac.uk).

Received 24 April; accepted 24 August; published online 24 September 2006; corrected after print 14 March 2007; doi:10.1038/ng1890

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Table 1 Association of SNPs spanning the CFH region with AMD
SNP number 1 2 3 4 5 SNP name rs1292487 rs512900 rs7524776 rs529825 rs800292 rs1329424 rs1061147 rs1061170 rs10801555 rs2019727 rs2019724 rs203685 rs1831281 rs2274700 rs6677604 rs3753396 rs419137 rs2284664 rs1065489 rs10801560 rs460897 rs432007 rs438781 rs408519 rs6428372 rs10922147 rs1971579 rs4085749 rs10922152 rs5998 CFHR2 C140C F13B N602N CFH D936E CFH S1191L CFH A473A CFH Q672Q CFH I62V CFH A307A CFH Y402H Coding variant Case/control ratios 308: 38, 263: 77 344: 2, 338: 2 318: 28, 292: 48 309: 37, 263: 77 309: 37, 263: 77 191: 155, 130: 210 191: 155, 130: 210 192: 154, 130: 210 192: 154, 130: 210 303: 27, 272: 66 213: 133, 141: 199 212: 134, 141: 199 293: 37, 269: 69 280: 66, 205: 135 319: 27, 274: 66 279: 67, 278: 62 282: 64, 296: 44 307: 39, 271: 69 278: 68, 276: 64 307: 39, 274: 66 319: 27, 270: 68 211: 133, 145: 191 214: 132, 148: 192 212: 134, 147: 193 282: 64, 281: 59 296: 48, 261: 79 262: 84, 223: 117 298: 48, 263: 77 209: 137, 161: 179 184: 162, 199: 141 w2 16.7 0.0 6.3 17.7 17.7 19.8 19.8 20.5 20.5 17.9 27.7 26.9 10.6 35.2 19.7 0.1 4.0 10.5 0.1 8.8 21.7 22.5 23.1 22.4 0.2 9.7 8.5 8.9 11.8 2.0 P value 4.33 105 0.99 0.01 2.61 105 2.61 105 8.47 106 8.47 106 5.96 106 5.96 106 2.29 105 1.41 107 2.12 107 0.0011 2.92 109 8.97 106 0.71 0.046 0.001 0.78 0.003 3.22 106 2.07 106 1.54 106 2.26 106 0.70 0.0018 0.0035 0.0029 0.0006 0.16

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6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

P values were generated using a w2 test for association of allele frequencies in 173 individuals with AMD and 170 controls.

We sought to identify the basis of haplotype 5 associated with decreased risk of AMD. Examination of quality scores for genotyping rs460897 (reported to encode S1191L within the nal exon of CFH) suggested that the assay may have measured a copy number variant rather than the expected SNP. Specically, we hypothesized that the assay might target a deletion of the nal exon of CFHR1, which differs from the nal exon of CFH only at the position of this putative SNP and at one other base. We selected gene-specic primers to sequence these exons in DNA from individuals homozygous for each of the ve haplotypes in block 2 who were also homozygous for the strongly associated haplotype in adjoining block 3. We did not nd any variation in exon 23 of CFH in association with any haplotype; hence, rs460897 does not represent a true SNP. Exon 6 of CFHR1 did not amplify in homozygotes for the protective haplotype 5 (Fig. 2a,b). In all other haplotypes, exon 6 of CFHR1 differed from CFH at the expected sites and also showed haplotype-specic variation at SNPs rs4320 (906G-T) and rs414628 (942A-T). All 28 samples sequenced from heterozygous individuals carrying one copy of haplotype 5 seemed to be homozygous at all CFHR1 exon 6 SNP sites, of which ten were homozygous for a rare allele. We conrmed deletion in haplotype 5 homozygotes by coamplication using primers that annealed to sites common to both CFH and CFHR1. Sequencing of PCR products showed that haplotype 5 amplied only CFH, whereas all other haplotypes amplied both genes, uncovering pseudoSNPs at all sites differing between CFH and CFHR1 (Fig. 2c). Similarly, we uncovered deletion of CFHR1 intron 4 by coamplication using

primers common to CFH intron 21 and CFHR1 intron 4, which amplied products of 324 and 380 bp, respectively (Fig. 2b). The deletion also removed CFHR3 from haplotype 5, as indicated by amplication only of CFHR4 sequence in homozygotes using primers specic to both exon 6 of CFHR3 and CFHR4 anking a region of incomplete homology (Fig. 2c). We used multiplex ligation-dependent probe amplication7 to measure copy number of CFH exon 23 and CFHR1 exon 6. This relatively new technique involves hybridization of pairs of adjoining probes to genomic DNA followed by ligation, which is achieved only if both are perfectly hybridized at the adjacent ends. Our assays centered on CFH 3572C and CFHR1 869T, and the hybridizing portions of the gene-specic probes varied only at one key base. The copy number of CFH exon 23 remained constant in male and female DNA samples from individuals carrying 0, 1 or 2 copies of haplotype 5 (who had copy numbers of 1.00, 1.04 and 1.06, respectively) when referenced to an autosomal marker in exon 9 of MORF4L1 and an X-linked marker in exon 6 of BCAP31, which was corrected for sex in males. As expected, the copy number of CFHR1 dropped from 1 to 0.44 and from 1 to 0 in heterozygotes and homozygotes for haplotype 5, respectively. Protein blot analysis of serum samples from individuals homozygous for each haplotype conrmed the absence of CFHR1 and CFHR3 protein in haplotype 5 homozygotes (Fig. 2d). We did not nd deletions on other haplotypes. The deletion on haplotype 5 measured 84,682 bases in length and occurred between two virtually identical 29-kb segments of

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Table 2 Association of CFH and related gene haplotype blocks with AMD
Haplotype Block 1 Markers 39 TCGAACA TCGCCTG CTACCTG TTACCTG 55.2 33.8 8.1 2.6 18.3 43.0 19.5 11.3 7.8 61.3 19.5 11.3 7.6 61.5 18.5 13.9 5.8 38.2 39.1 14.1 8.5 12.9 28.5 18.2 20.3 19.4 43.2 17.2 19.4 19.8 42.0 17.0 22.6 17.4 1.99 0.80 0.54 0.29 1.51 1.89 1.08 0.50 0.35 2.08 1.16 0.53 0.33 2.22 1.10 0.55 0.29 19.8 2.1 6.3 11.5 3.7 15.6 0.2 10.3 19.5 22.5 0.6 8.6 21.8 26.1 0.2 8.9 22.5 8.47 106 0.15 0.012 0.0007 0.053 7.72 105 0.68 0.0013 1.03 105 2.14 106 0.45 0.003 2.96 106 3.25 107 0.62 0.003 2.14 106 Percentage in affected individuals Percentage in controls Odds ratio w2 P value

2007 Nature Publishing Group http://www.nature.com/naturegenetics

Block 2 Markers 1118

1: AGGCGACG 2: AGGCGAAG 3: GTGCGGAG 4: GTATGAAA 5: GTGTAAAG

Block 3 Markers 2024

CATAC CAGTT AAGTT CGGTT

Block 4 Markers 2528

ACAC CCCC AGAT ACCC

An odds ratio 41 indicates a detrimental haplotype, and o1 indicates a protective AMD haplotype.

duplication in the physical map of the chromosome8 (marked 1 and 1 in turquoise in Figs. 1a and 2a). The position of the deletion was mapped by amplifying regions of slight variation between the duplicated regions and sequencing to check which base was retained in homozygotes for haplotype 5 (Fig. 2a,c). A 2.8-kb junction fragment was identical in size to that found in nondeleted haplotypes; hence, both breakpoints map to the same point within a 1,618-bp region of perfect homology centered B6,400 bp after CFH and CFHR1 (Supplementary Fig. 2 online). CFH and related proteins are important regulators of complement activity. The CFH gene cluster is responsible for numerous alternatively spliced transcripts and proteins. Their high homology makes them extremely difcult to study, and the contribution of each is poorly understood. CFH and CFHR1 are known to be present in the circulation at high levels and to act as cofactors for factor Imediated degradation of C3b9,10. Notably, our data show that CFHR3 and CFHR4A (the long CFH a form of CFHR4) (ref. 11) are also present in serum at comparable levels. It has not yet been possible to assess the relative impor-

tance of deletion of CFHR1 and CFHR3 in contributing to the protective nature of haplotype 5; however, the products of both genes are present in the circulation where they have the potential to compete with CFH for C3 binding. Mutations in the early exons of CFH that cause hemolytic uremic syndrome (HUS; see http:// www.FH-HUS.org; OMIM #235400) tend to affect CFH protein secretion rather than function. Although only semiquantitative, our protein blot data suggest that haplotypes 1 and 2, which are associated with the harmful C allele of Y402H, may have relatively less serum CFH in comparison with CFH-related proteins. A possible mechanism for this is through the close association of the C allele with the G allele of SNP rs10922094, which may permit some alternative splicing of a new exon containing an immediate stop codon. This putative exon is distinct from exon 10, which is alternatively spliced as the nal exon of the shorter CFH b transcript.
CFHR3 CFHR1 CFHR4 CFHR2 CFHR5

Figure 1 Haplotype block structure in CFH and CFH-related genes. (a) Duplicated regions that share orientation and 496% homology are shown below the genomic structure of genes. Many other regions of 80%95% homology are not marked. The haplotype block structure of markers used in this study is shown in blue. (b) Haplotypes of SNPs are shown in blocks with overall frequencies (in affected individuals and controls) and connections from one block to the next. In the crossing areas, a value of multiallelic D is shown. This reects the level of recombination between two adjacent blocks. The black box outlines the protective haplotype 5 in this block.

1 Block 1
rs7524776 rs529825 rs800292 rs1329424 rs1061147 rs1061170 rs10801555

2 Block 3

2 Block 4
rs10801560 rs460897 rs432007 rs438781 rs408519

3 50 kb

Block 2

0.16 0.47 0.36 0.11 0.06 0.89 Haplotype Block 1 Haplotype Block 2 0.36 0.19 0.16 0.14 0.91 Haplotype Block 3 0.52 0.18 0.15 0.14 0.89 Haplotype Block 4 0.52 0.18 0.18 0.11

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rs6428372 rs10922147 rs1971579 rs4085749

rs2019724 rs203685 rs1831281 rs2274700 rs6677604 rs3753396 rs419137 rs2284664

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(170) were age matched with no evidence of early macular disease in either eye (stages 0 or 1) and were recruited through random sampling of older adults (65 years and above) from a populationbased register. All were recruited in Northern Ireland, UK and were of European descent. The replication group of 192 subjects matched the above criteria for subjects. The replication group of controls consisted of 192 elderly UK individuals of European descent who had no eye examination. DNA extraction, genotyping and sequencing. DNA was extracted from peripheral blood by standard methods. High-throughput SNP genotyping was outsourced using Illumina bead technology based on multiplex PCR and primer extension as part of a larger project. Additional SNPs were typed in-house using multiplex PCR followed by multiplex SNaPshot (ABI) technology. Primers were designed using Primer Detective (Clontech). Primer sequences for specic and nonspecic amplication of CFH and related genes are listed in Supplementary Table 3 online. Sequencing was performed using ABI dye terminator chemistry v3 with analysis on an ABI3100 genetic analyzer. We used Sequencher software (Genecodes) to compare DNA sequences.

a
CFH CFHR3 CFHR1
Deleted in haplotype 5

c
CFH CFHR1 1 CFHR3 CFHR4

rs10801560

2007 Nature Publishing Group http://www.nature.com/naturegenetics

rs432007 rs438781 rs408519

Dup1 Dup1

b
CFH exon 23

d
CFH

Anti-CFHR1 kDa 175 83 62 47.5 32.5 25

Anti-CFHR3

CFH CFHR4

CFHR1 exon 6
CFHR1

CFHR3

CFHR1 intron 4 CFH intron 21 1 2 3 4 5

1 2 3 4 5

Figure 2 Deletion of CFHR3 and CFHR1 in homozygotes for haplotype 5. (a) Map of the terminal seven exons of CFH and all six exons of CFHR3 and CFHR1. The 29-kb region of duplication between CFH and CFHR1 is shown, and the region deleted in haplotype 5 is boxed in red. Haplotype block 3 and the position of SNPs within this block are indicated. rs460897 is omitted because of lack of evidence that this represents a true SNP. (b) DNA from homozygotes for each of the ve haplotypes (15) of block 2 was amplied using primers specic for CFH exon 23, primers specic for CFHR1 exon 6 and primers that amplied different-sized products from CFHR1 intron 4 and CFH intron 21. Haplotype 5 did not amplify from CFHR1. (c) Pairs of sequences show one homozygote for haplotype 5 (lower trace) and a homozygote representative of all other haplotypes (upper trace) of PCR products coamplifying from two imperfectly duplicated loci. PseudoSNPs are present in all haplotypes except haplotype 5, which did not show amplication from CFHR1 or CFHR3. The bottom pair of traces delineates the breakpoint region of the deletion on haplotype 5, where the sequence of haplotype 5 amplies from adjacent duplicated (Dup) regions 1 and 1. (d) Protein blotting of serum from homozygotes for haplotypes 1 to 5 with polyclonal antisera raised against CFHR1 and CFHR3.

It is an attractive hypothesis that CFH produced from the fulllength transcript is benecial and other CFH-related proteins interfere with regulation of complement activity. Much work is required to unravel the complexity of the transcripts and proteins arising from this highly duplicated gene cluster. The prevalence of AMD is growing in parallel with the increasing longevity of the population, and with no effective treatment, the disease presents a major challenge. Identication of CFH as a major susceptibility gene in 2005, closely followed by a second locus (LOC387715, or PLEKHA1)12,13 on chromosome 10, were important breakthroughs in AMD research. Starting to resolve the genetic basis for the different haplotypic effects and the complex roles of the CFH-related genes in predisposition to AMD may shed some light on its etiology and in the future might present useful therapeutic targets for gene silencing. METHODS
Study populations. This study was approved by the Research Ethics Committee of the Queens University of Belfast. Informed consent was obtained from all subjects involved, in compliance with all principles of the Helsinki Accord. Grading of digital fundal color photographs was according to the classication of mutually exclusive stages of age-related maculopathy (ARM)14 based on the international classication system for ARM15. Under this system, stages 2 and 3 correspond to early ARM and stage 4 to AMD. Our study subjects with AMD (173) had end-stage neovascular AMD (stage 4) in at least one eye. They were selected from Ophthalmology Clinics in the Royal Victoria Hospital, Belfast, UK, and had an average age of 76 years. The controls in our main study

Statistical methods. Genotype data from affected individuals and controls were loaded into Haploview16 (http://www.broad.mit.edu/mpg/haploview/) in linkage format to generate allele and haplotype numbers, ratios and P values based on the w2 test for association of allele or haplotype frequencies. Logistic regression analysis was performed using Stata v8 to assess further the relative contributions of rs1061170 (Y402H) and the presence or absence of deletion of CFHR1 and CFHR3 using Stata on our main data set of 173 AMD cases and 170 controls with no signs of macular disease. The variables representing His402 and the deletion haplotype 5 were coded 0, 1 or 2 depending on the number of copies carried. Multiplex ligation-dependent probe amplication (MLPA). This was performed on 100 ng of DNA using buffers, enzymes and PCR primers from MRC Holland according to their protocol. Hybridizing probe sequences X-CFH1191C or X-CFHR1-1191T were used in separate reactions with a common PHO-labeled oligonucleotide, CFH1191-Y. Controls using MORF4L1 and BCAP31 were included in all reactions. Analysis was based on peak heights and correlated well with peak areas. All MLPA oligonucleotide sequences are available in Supplementary Table 3.

Protein blotting. Serum samples were separated by polyacrylamide gel electrophoresis, transferred on to nylon membrane and protein blotted using polyclonal antisera raised against CFHR1 and CFHR3 using standard methods. Accession codes. GenBank: CFH, NM000186; CFHR1, BC016755; CFHR2, BC022283; CFHR3, AK124051; CFHR4, BC074957. CFH exons are numbered to include exon 10, which is alternatively spliced into the shorter transcript of this gene. Numbering of transcripts starts from the initial ATG.
Note: Supplementary information is available on the Nature Genetics website. ACKNOWLEDGMENTS We thank D. McGibbon for assistance with DNA extraction; J. Silvestri, V. McConnell, G. Wright, G. Meenagh and C. Benson for blood samples from their patients; P. Zipfel for antisera for use in protein blotting; C. Cardwell for statistical help and Illumina for discussions on choice of SNPs for typing. This work was supported by The Health Foundation.

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AUTHOR CONTRIBUTIONS A.E.H., N.O. and H.E. designed the SNP association study, A.E.H. and N.O. analyzed SNP data, T.G. and M.D.-T. performed protein blotting experiments, U.C. provided clinical information and patient samples and A.E.H. performed all other experiments and wrote the paper. COMPETING INTERESTS STATEMENT The authors declare competing nancial interests (see the Nature Genetics website for details).
Published online at http://www.nature.com/naturegenetics Reprints and permissions information is available online at http://npg.nature.com/ reprintsandpermissions/
1. Mullins, R.F., Aptsiauri, N. & Hageman, G.S. Structure and composition of drusen associated with glomerulonephritis: implications for the role of complement activation in drusen biogenesis. Eye 15, 390395 (2001). 2. Klein, R.J. et al. Complement factor H polymorphism in age-related macular degeneration. Science 308, 385389 (2005). 3. Edwards, A.O. et al. Complement factor H polymorphism and age-related macular degeneration. Science 308, 421424 (2005). 4. Haines, J.L. et al. Complement factor H variant increases the risk of age-related macular degeneration. Science 308, 419421 (2005). 5. Hageman, G.S. et al. A common haplotype in the complement regulatory gene factor H (HF1/CFH) predisposes individuals to age-related macular degeneration. Proc. Natl. Acad. Sci. USA 102, 72277232 (2005). 6. Zareparsi, S. et al. Strong association of the Y402H variant in complement factor H at 1q32 with susceptibility to age-related macular degeneration. Am. J. Hum. Genet. 77, 149153 (2005). 7. Schouten, J.P. et al. Relative quantication of 40 nucleic acid sequences by multiplex ligation-dependent probe amplication. Nucleic Acids Res. 30, e57 (2002). 8. Heinen, S. et al. De novo gene conversion in the RCA gene cluster (1q32) causes mutations in complement factor H associated with atypical hemolytic uremic syndrome. Hum. Mutat. 27, 292293 (2006). 9. Reid, K.B. et al. Complement system proteins which interact with C3b or C4b; a superfamily of structurally related proteins. Immunol. Today 7, 230234 (1986). 10. DiScipio, R.G. Ultrastructures and interactions of complement factors H and I. J. Immunol. 149, 25922599 (1992). 11. Jozsi, M. et al. FHR-4A: a new factor H-related protein is encoded by the human FHR-4 gene. Eur. J. Hum. Genet. 13, 321329 (2005). 12. Jakobsdottir, J. et al. Susceptibility genes for age-related maculopathy on chromosome 10q26. Am. J. Hum. Genet. 77, 389407 (2005). 13. Rivera, A. et al. Hypothetical LOC387715 is a second major susceptibility gene for age-related macular degeneration, contributing independently of complement factor H to disease risk. Hum. Mol. Genet. 14, 32273236 (2005). 14. van Leeuwen, R., Klaver, C.C., Vingerling, J.R., Hofman, A. & de Jong, P.T. The risk and natural course of age-related maculopathy: follow-up at 6 1/2 years in the Rotterdam study. Arch. Ophthalmol. 121, 519526 (2003). 15. The International ARM Epidemiological Study Group. An international classication and grading system for age-related maculopathy and age-related macular degeneration. Surv. Ophthalmol. 39, 367374 (1995). 16. Barrett, J.C., Fry, B., Maller, J. & Daly, M.J. Haploview: analysis and visualization of LD and haplotype maps. Bioinformatics 21, 263265 (2005).

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Corrigendum: A common CFH haplotype, with deletion of CFHR1 and CFHR3, is associated with lower risk of age-related macular degeneration
Anne E Hughes, Nick Orr, Hossein Esfandiary, Martha Diaz-Torres, Timothy Goodship & Usha Chakravarthy Nat. Genet. 38, 11731177 (2006); published online 24 September 2006; corrected after print 14 March 2007 In the version of this article initially published, the G and A alleles of rs1831281 in Figure 1 should be reversed, and the block 2 haplotypes in Figure 1, Table 2 and Supplementary Table 2 should be corrected to 1:AGGCGACG, 2:AGGCGAAG, 3:GTGCGGAG, 4:GTATGAAA and 5:GTGTAAAG. The error has been corrected in the HTML and PDF versions of the article. 2007 Nature Publishing Group http://www.nature.com/naturegenetics