INTRODUCTION

were further adjusted for the overall genomic inflation factor (l¼ 1.019) (Supplementary Material, Fig. S1). A flow chart of the consecutive analysis steps is depicted in Figure 1.

Worldwide, human life expectancy has increased remarkably over the last two centuries (1), although the healthy life expect- ancy lags behind. Citizens of the European Union, for example, spend only 75–80% of their lifespan in good health (2). Families in which longevity clusters form an exception in this sense, by showing beneficial or ‘youthful’ profiles for many metabolic and immune-related parameters (3–7) and a low prevalence of common diseases from middle age onwards (5,8,9). Therefore, the genome of long-lived individuals is investigated to identify variants that promote healthy aging and protect against age-related disease. This is a major challenge because the genetic component of lifespan variation in the population at wait large has been estimated to be only 25% (10,11) and is assumed to be determined by many, still uncharacterized, genes (12,13). Genetic influences on human longevity are expected to reflect longevity assurance mechanisms acting across species (14), as well as more heterogeneous population- specific effects. Although numerous genome-wide association studies (GWAS) have successfully identified loci involved in common, age-related diseases (15), the corresponding suscepti- bility loci do not explain the genetic component of human lon- gevity (16). GWAS for human longevity have thus far failed to identify genome-wide significant loci, besides the well-known TOMM40/APOE/APOC1 locus (17–19).

years (n ¼ 7729) showed genome-wide significant association with survival into old age at one locus, the previously identified TOMM40/APOE/APOC1 locus (17,21) (rs4420638 (G); odds ratio (OR) ¼ 0.71, P ¼ 6.14 × 10219; Table 1). No gender- dependent effects were observed in the sex-stratified analysis ≥ of the cases aged 85 years (Supplementary Material, Table S4). ≥ The discovery-phase meta-analysis of the cases aged 90 years (n ¼ 5406) showed a similar result, i.e. the TOMM40/ APOE/APOC1 locus was the only genome-wide significant locus (OR ¼ 0.64, P ¼ 4.09 × 10221; Fig. 2 and Table 2). The regional association plot and forest plot for the TOMM40/ APOE/APOC1 locus are depicted in Figures 3 and 4, respective- ly. Although several SNPs on chromosome 19q13.32, which are in moderate linkage disequilibrium (LD) with rs4420638, show additional association with survival into old age, meta-analysis conditional on rs4420638 showed no independent associations amongtheseSNPs(SupplementaryMaterial,FigS2andTableS3).

Replication

In addition to the TOMM40/APOE/APOC1 locus, we found eight loci that showed suggestive evidence for association in ≥ the discovery-phase meta-analysis of cases aged 85 years (P ≤ 1 × 1025; Table 1), whereas six additional SNPs met this ≥ criterion in the meta-analysis of cases aged 90 years (Table 2). The most or (when not successfully measured) second most significant SNPs from these 14 loci and the TOMM40/APOE/APOC1 locus were taken forward for replica- ≥ tion in 13 060 cases aged 85 years (of which 7330 were also ≥ 90 years) and 61 156 controls from 6 additional studies. In the joint analysis of the discovery and replication phase of the ≥ cases aged 85 years (9 loci), the TOMM40/APOE/APOC1 locus remained the only genome-wide significant locus (Table 1). The joint analysis of the discovery and replication ≥ phase of the cases aged 90 years (12 loci), however, showed an additional genome-wide significant locus, rs2149954 (T), on chromosome 5q33.3 (OR ¼ 1.10, P ¼ 1.74 × 1028; Table 2). Although the association of this SNP with survival up to 85 years is not genome-wide significant (OR ¼ 1.07, P ¼ 4.34 × 1026; Table 1), the locus likely affects survival from middle age onwards. The regional association plot (based on the discovery phase only) and forest plot of this locus are depicted in Figures 3 and 4, respectively. Conditional analysis of rs4420638 in the discovery phase studies showed that the as- sociation of rs2149954 (T) with survival is independent of the TOMM40/APOE/APOC1 locus (P ¼ 7.20 × 1026 instead of P ¼ 5.98 × 1026 in the analysis of survival up to 85 years).

In this paper, we conducted a large genome-wide association meta-analysis of human longevity in 14 studies with long-lived ≥ cases ( 85 years) and younger controls (,65 years) from European descent. In addition, we performed a subset analysis ≥ incases aged 90 years. The novel longevity locus we identified wastestedforassociationwithprospective(cause-specific) mor- tality in a meta-analysis of 11 European cohorts and examined for association with various metabolic traits that may explain the mechanism by which the locus contributes to survival to high ages.

RESULTS

Genome-wide association analysis