In addition, we incorporated gene annotation informa- tion, a technique that has successfully been applied in the field of aging research (de Magalhaes et al., 2009a, 2010). Protein ANalysis THrough Evolutionary Relationships (PANTHER; Mi et al., 2007; Thomas et al., 2003) and Ingenuity Pathway Analysis (IPA) (www.ingenuity.com) were used for identification and classification of networks, pathways, biological processes, and molecular functions of the genes identified in this study. For both phenotypes we generated lists of candidate genes. These genes were the closest reference genes to the SNPs associated with the outcome at p  1 oN 10(cid:3)3. PANTHER compares these gene lists to the reference list using the binomial test for each molecular function, biological process, or pathway term.

Observed (-logP)Observed (-logP) 43  Expected (-logP)    Expected (-logP)

3. Results

death adjusted for baseline age and sex in participants of European origin, 55 years of age or older from 9 longitu- dinal cohort studies participating in the CHARGE Consor- tium (Psaty et al., 2009). In total, we observed 8444 deaths (mean age at death: 81.1, SD 8.4) in 25,007 participants (55% female) after an average follow-up of 10.6 (SD 5.4) years. Descriptive characteristics of participants and Man- hattan plots showing genome wide p-values for association are displayed in the Supplementary data (Supplementary Fig. S1, and Supplementary Tables S3 and S4). The quan- tile-quantile plot (Q-Q plot) of observed versus expected p-values showed only a small deviation from the null hy- pothesis, indicating no significant population stratification   (Fig. 1a, 1.066). Although there were no genome- GC wide significant findings (p   5 _ 10(cid:3)8), 14 independent SNPs were associated with time to death at a suggestive threshold of p << 1  10(cid:3)5 (Table 1). Among these SNPs, rs4936894 (chromosome 11, near the von Willebrand factor A domain containing 5A gene [VWA5A]) had the strongest association with time to death (p  3.4 4ads| xX 10(cid:3)7). We sought replication for 5 of the 14 top SNPs with the strongest association with time to death in 4 independent samples    (n 10,411, deaths 1295) of the same ancestry. None of the SNPs were consistently associated with time to death at << a nominally significant level of p 0.05 across all replica- tion samples (Supplementary Tables S5–S8). In the com- bined meta-analysis of the discovery and replication studies only rs1425609 in the vicinity of otolin-1 (OTOL1) showed a stronger association (1.61 “J ™ 10(cid:3)6).

Likewise, no genome-wide significant findings were identified in the time to event analysis following 16,995 participants free of disease at baseline and registering 7314 events over an average of 8.8 (SD 5.7) years of follow-up (Table 2). Events included incident myocardial infarction, heart failure, stroke, dementia, hip fracture, and cancer or ane  death. The Q-Q plot (Fig. 1a, 1.019) showed no GC evidence of inflation of type I error. In total, there were 8 independent SNPs associated with event-free survival at p er 10(cid:3)5. The SNP with the strongest association was rs10412199 (chromosome 19, p  3.02   10(cid:3)6), which is in close proximity to ataxia, cerebellar, Cayman type (AT- CAY). Additional descriptive information including defini- tions of each event and association results with p co‘ 10(cid:3)4 are provided in Supplementary Figure S2, and Supplemen- tary Tables S9–S12.

Fig. 1. (a) Quantile-quantile (Q-Q) plot after meta-analysis for time to death. (b) Quantile-quantile (Q-Q) plot after meta-analysis for time to event.

the overlap between their association results (Table 3). In- terpretation of the overlap between the phenotypes requires caution as both phenotypes are correlated, nevertheless it helps to focus on specific loci and put them into the context of aging. From the 14 loci passing the prespecified, sugges- tive threshold of p r< 1  10(cid:3)5 in the time to death analysis, 5 had corresponding SNPs within 500 kilo base pairs dis- tance, in linkage disequilibrium (LD; r2  0.1) associated with p << 1  10(cid:3)4 and the same overall direction of the effect in the time to event analysis. These 5 regions were in the vicinity of the following genes: OTOL1 (3q26.1), bridg- ing integrator 2 (BIN2, 12q13), ATG4 autophagy related 4 homolog C (ATG4C, 1p31.3), origin recognition complex,

As both phenotypes may provide different but compli- mentary information about the aging process, we evaluated

Table 1

Top 14 SNPs (p-value   10-5) for time to death ranked by p-value, from meta-analysis of 9 cohortsa

Number SNP Chr Position Closest reference Distance from Coded Noncoded Frequency HR p-value Study effect gene closest gene allele allele coded allele direction b 1 rs4936894 11 123522703 VWA5A 123 A G 0.226 1.11 3.38E-07 +t4te¢t-+ 224 2 rs1425609 3 164164689 OTOL1 1,460,265 A G 0.381 0.92 1.46E-06 —------ 399 3 rs766903 12 49990101 BIN2 14,104 A G 0.834 0.90 1.61E-06 —-------(cid:4)---- 7 4 rs12042640 1 63139384 ATG4C 36,747 T C 0.284 1.09 1.71E-06  +4+++-+-+- 19 5 rs17149227 7 75073485 HIP1 72,141 T G 0.959 0.79 3.56E-06 (cid:3)??---------(cid:4)-? 0 6 rs3128591 9 136741940 COL5A1 68,468 A G 0.754 0.92 3.64E-06 —------ 20 7 rs11582903 1 87618642 LMO4 34,804 A C 0.150 1.12 3.94E-06 ttt bat  38 8 rs4850695 2 196861504 HECW2 89,283 A G 0.766 1.09 4.62E-06 FN++t+t4++4+ 95 9 rs10259086 7 103680248 ORC5L 44,549 T G 0.686 1.08 5.16E-06 asFHP E+ HTH 72 10 rs2769255 1 41017941 KCNQ4 4329 T C 0.374 1.08 5.17E-06 Fst+t4t+-4++ 95 11 rs17291546 6 2660681 LOC340156 35,472 A G 0.957 0.82 7.65E-06 (cid:3)?------- 8 12 rs12606100 18 69102967 NETO1 417,177 T C 0.202 1.11 8.72E-06   4 13 rs1274214 11 122979741 GRAMD1B 18,987 T C 0.500 0.93 8.87E-06 —------ 42 14 rs10811679 9 2224701 SMARCA2 41,080 T C 0.330 1.08 9.53E-06 ++t+t4++4+  37