Abstract
Here we describe the insights gained from sequencing the whole genomes of 2,636 Icelanders to a median depth of 20×. We found 20 million SNPs and 1.5 million insertions-deletions (indels). We describe the density and frequency spectra of sequence variants in relation to their functional annotation, gene position, pathway and conservation score. We demonstrate an excess of homozygosity and rare protein-coding variants in Iceland. We imputed these variants into 104,220 individuals down to a minor allele frequency of 0.1% and found a recessive frameshift mutation in MYL4 that causes early-onset atrial fibrillation, several mutations in ABCB4 that increase risk of liver diseases and an intronic variant in GNAS associating with increased thyroid-stimulating hormone levels when maternally inherited. These data provide a study design that can be used to determine how variation in the sequence of the human genome gives rise to human diversity.
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Acknowledgements
We thank all the participants in this study. This study was performed in collaboration with Illumina.
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D.F.G., H. Helgason, S.A.G., F.Z., D.O.A., O.T.M., G. Masson, A.H., P.S. and K.S. wrote the initial draft of the manuscript. D.F.G., H. Helgason, S.A.G., F.Z., A.O., G. Magnusson, B.V.H., E.H., G.T.S., S.N.S., M.L.F., A.K., G. Masson and P.S. analyzed the data. D.F.G., H. Helgason, S.A.G., F.Z., A.G., S.B., H.G. and G. Masson created methods for analyzing the data. S.N.S., H. Holm, J.S., H.T.H., H.J. and O.T.M. performed the experiments. H. Holm, G.S., G.T., J.T.S., S.G., G.B.W., T.R., B.T., E.S.B., S.O., H.T., T.S., T.S.G., A.T., J.G.J., A.S., G.B., J.J.J., O.T., P.L., G.I.E., O.S., I.O. and D.O.A. collected the samples and information. D.F.G., D.O.A., G. Masson, U.T., A.H., P.S. and K.S. designed the study.
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The authors affiliated with deCODE Genetics are employed by the company, which is owned by Amgen, Inc: D.F.G., H. Helgason, S.A.G., F.Z., A.O., A.G., G. Magnusson, B.V.H., E.H., G.T.S., S.N.S., M.L.F., H. Holm, J.S., H.T.H., H.J., S.G., G.B.W., T.R., A.S., G.B., H.G., O.T.M., A.K., G. Masson, U.T., A.H., P.S. and K.S.
Integrated supplementary information
Supplementary Figure 4 Distribution of the number of observed alleles in 2,636 sequenced Icelanders by impact class.
Shown are the proportions of variants for which the minor allele was seen one to six times (MAF ≤ 0.11%).
Supplementary Figure 5 Comparison of imputed and chip genotypes.
Shown is the fraction of the 28,204 SNPs identified in exons and splice regions and present on SNP chips that have r2 > 0.8, 0.9 and 0.99 between the imputed and chip genotypes as a function of their derived allele frequency (DAF).
Supplementary Figure 6 The five pedigrees containing the eight homozygous carriers of c.234delC in MYL4.
Symbols for homozygous carriers are colored black. Symbols for deceased individuals are stricken through with a forward-leaning line. Symbols for individuals who have not been genotyped directly are stricken through with a backward-leaning line. Under each individual are up to five lines containing information about the individual. First appear an identifier, consisting of a pedigree name (f1–f5), the generation of the individual in roman numerals and an enumerator within the generation. Second appear the individual’s year of birth and, if appropriate, the individual’s year of death. Third appears the individual’s c.234delC genotype, where D and W denote directly genotyped deletion and wild-type alleles, respectively, and d and w denote in silico genotypes inferred from the genotypes of relatives. The order of the alleles indicates the parent of origin, where the first allele comes from the father and the second allele comes from the mother, except for the three cases for whom parent of origin could not be assigned: f2-I:1, f2-I:2 and f3-I:2. Fourth appear an indication of whether the individual has been diagnosed with AF and the age at onset after the @ sign. Fifth appear the presence of other relevant phenotypes: sick sinus syndrome (SSS), pacemaker implantation (PM) and sudden cardiac death (SCD).
Supplementary Figure 7 The transmission of chromosome 17 through pedigree f-2.
The transmission of the founding couple of pedigree f-2 can be reconstructed on the basis of the expected values for meiotic transmissions of chromosome 17. The horizontal red lines indicate the position of c.234delC in MYL4, and the small red square surrounding the line indicates the region around c.234delC shared identically by decent by the founding couple. The length of this interval is estimated to be 3.3 cM. The first and last 10 cM of the chromosome have been truncated. The sisters f2-II:6 and f2-II:9 are imputed to be carrying c.234delC on their paternal chromosome on the basis of the chromosomal region around the deletion having been transmitted to their children (dark blue). There is no clear transmission of either sister’s maternal chromosomal region around the deletion to one of her children (although f2-III:3 may have inherited her mother’s paternal chromosome, but a crossover occurred in the region around c.234delC where f2-III:3 is homozygous). However, for both sisters, the maternal chromosome carrying the deletion (light blue) was transmitted to an offspring at regions on both sides of MYL4 (to f2-III:2 and f2-III:4) such that, unless a double crossover occurred around MYL4, they both carry c.234delC on their maternal chromosome.
Supplementary Figure 8 The families of the BVVL cases.
Shown are birth years and genotypes at the SLC52A2 mutation, where W denotes the wild-type allele and M denotes the mutated allele. Symbols for cases are colored black, and the symbols corresponding to the two siblings of case 4 who died early are colored gray. A forward slash indicates that the individual is deceased, and a backward slash indicates that an SLC52A2 genotype is not available for that individual.
Supplementary Figure 9 The effect of the filtering steps on the number of sequence variants that are candidates for causing BVVL syndrome in the two sisters.
The occurrence of a rare syndrome such as BVVL in two sisters suggests that it is caused by a rare genotype with high penetrance. We therefore restricted our search to LoF and MODERATE-impact variants. The sisters are affected but neither parent is, which suggests an autosomal recessive mode of inheritance. Allelic frequency over 2% would dictate a homozygous frequency of over 1 in 2,500, which would be too high for BVVL syndrome. This brought the number of potential variants down to 3 from 147. This would not have been possible using non-Icelandic resources such as ESP, as 4 of the 147 variants are not present in the database. We note that crude filtering, such as removing all variants present in public databases, would result in removing the causative sequence variant. This left us with three correlated MODERATE-impact variants on chromosome 8q24.3: p.Leu339Pro (rs148234606) in SLC52A2, p.Gln931Arg in OPLAH and c.2982C>T in CPSF1. No one was imputed to be homozygous for the SLC52A2 variant in the set of additional chip-typed Icelanders, whereas 5 and 19 Icelanders were imputed to be homozygous for the OPLAH and CPSF1 SNPs, respectively. No early deaths were reported among these homozygous carriers, and the oldest homozygous carriers reached ages 77 and 89 years for OPLAH and CPSF1, respectively, which is inconsistent with diagnosis of BVVL, as only 1 of 77 reported BVVL syndrome cases has lived past 60 years36,37,38,39,40,41,42,43,44.
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Supplementary Figures 1–9, Supplementary Tables 1–15 and Supplementary Note. (PDF 1678 kb)
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Gudbjartsson, D., Helgason, H., Gudjonsson, S. et al. Large-scale whole-genome sequencing of the Icelandic population. Nat Genet 47, 435–444 (2015). https://doi.org/10.1038/ng.3247
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DOI: https://doi.org/10.1038/ng.3247
