Table 2 Association statistics for 36 haplotype-tagging single-nucleotide polymorphisms (htSNPs)in FOXO1,FOXO4, and FOXO6 in German female LLI and controls

minAF minAF Gene Chr. SNP bp LLI controls P allele P geno OR (95% C.I.) FOXO1 13 rs2701893 40 023 636 0.345 0.353 0.617 0.348 0.97 (0.84–1.11) FOXO1 13 rs2721047 40 025 682 0.082 0.098 0.091 NA 0.82 (0.66–1.03) FOXO1 13 rs17446593 40 026 085 0.167 0.182 0.239 0.383 0.90 (0.76–1.07) FOXO1 13 rs2755209 40 035 804 0.369 0.376 0.654 0.109 0.97 (0.85–1.11) FOXO1 13 rs1078892 40 036 020 0.183 0.196 0.288 0.054 0.92 (0.78–1.08) FOXO1 13 rs2701859 40 039 232 0.247 0.269 0.130 0.078 0.89 (0.77–1.03) FOXO1 13 rs12865518 40 041 190 0.256 0.243 0.383 0.581 1.07 (0.92–1.24) FOXO1 13 rs2721069 40 041 720 0.309 0.323 0.367 0.303 0.94 (0.82–1.08) FOXO1 13 rs2755213 40 044 301 0.087 0.093 0.548 0.808 0.93 (0.75–1.17) FOXO1 13 rs2701880 40 048 505 0.058 0.070 0.121 NA 0.81 (0.62–1.06) FOXO1 13 rs2951787 40 059 770 0.382 0.372 0.513 0.806 1.05 (0.92–1.20) FOXO1 13 rs2984121 40 059 979 0.185 0.181 0.800 0.898 1.02 (0.86–1.21) FOXO1 13 rs2721044 40 060 225 0.307 0.328 0.170 0.135 0.91 (0.79–1.04) FOXO1 13 rs4943794 40 071 408 0.212 0.225 0.346 0.402 0.93 (0.79–1.08) FOXO1 13 rs1986649 40 076 824 0.212 0.227 0.277 0.413 0.92 (0.79–1.07) FOXO1 13 rs17630266 40 092 032 0.036 0.030 0.315 NA 1.20 (0.84–1.73) FOXO1 13 rs12876443 40 094 877 0.109 0.108 0.880 0.901 1.02 (0.82–1.25) FOXO1 13 rs7981045 40 107 236 0.278 0.266 0.435 0.696 1.06 (0.92–1.23) FOXO1 13 rs9603776 40 121 886 0.027 0.024 0.530 NA 1.14 (0.76–1.72) FOXO4 X rs12013673 70 233 824 0.432 0.428 0.800 0.574 1.02 (0.89–1.16) FOXO4 X rs5981072 70 236 267 0.298 0.296 0.927 0.982 1.01 (0.87–1.16) FOXO6 1 rs11585393 41 602 844 0.296 0.288 0.606 0.537 1.04 (0.90–1.20) FOXO6 1 rs7539614 41 603 634 0.443 0.437 0.707 0.924 1.03 (0.90–1.17) FOXO6 1 rs7547654 41 603 948 0.460 0.449 0.476 0.763 1.05 (0.92–1.19) FOXO6 1 rs11581271 41 607 648 0.395 0.408 0.446 0.445 0.95 (0.83–1.09) FOXO6 1 rs1317558 41 611 670 0.121 0.103 0.084 0.070 1.20 (0.98–1.47) FOXO6 1 rs1317557 41 611 929 0.028 0.025 0.541 NA 1.14 (0.76–1.71) FOXO6 1 rs6693260 41 612 082 0.168 0.179 0.388 0.591 0.93 (0.78–1.10) FOXO6 1 rs4660532 41 612 784 0.342 0.358 0.333 0.598 0.94 (0.82–1.07) FOXO6 1 rs6690527 41 614 573 0.286 0.283 0.830 0.405 1.02 (0.88–1.17) FOXO6 1 rs4660192 41 614 937 0.454 0.448 0.725 0.939 1.02 (0.90–1.17) FOXO6 1 rs11209971 41 616 880 0.281 0.279 0.887 0.990 1.01 (0.87–1.17)

Chr., chromosome; bp, base-position of NCBI build 36; minAF, minor allele frequency; LLI, long-lived individuals; P allele, P value obtained from allele-based case–control comparison using a chi square-test with 1 degree of freedom (df); P geno, P value obtained from genotype-based case–control comparison using a chi square-test with 2 df; OR, estimated odds ratio for minor allele (based on whole sample); CI, 95% confidence interval of OR; NA, genotype- based case–controls comparison was not conducted when there were fewer than five observations. Associated markers from the study of Li et al. (2009) are in bold.

Fig. 2), the data from the two populations were pooled. Also the combined sample with an a priori power of 84% did not confirm the previous finding of Li et al. (Table 3). Furthermore, in the German sample of female individuals (Table 4), we did not detect any association between the two haplotypes (ATG and CCG based on rs2755209, rs2755213, and rs17630266) and the longevity phenotype, as earlier reported in Chinese (Li et al., 2009).

The markers used for the FOXO1 haplotype analysis in the Leiden 85+ study (Kuningas et al., 2007) were also sub- jected to a haplotype analysis in our sample (rs2721069, rs4943794, and rs7981045; Fig. 1A). The frequency of the observed mortality haplotype in the Dutch sample (TCA) was not significantly different in German LLI compared with

Recently, association signals for the FOXO1 SNPs rs2755209 and rs2755213 were reported in two samples of female Han Chinese (Li et al., 2009). In Germans, the frequencies of the polymorphisms did not differ significantly between the LLI and the young controls, neither for the complete sample (Table 1) nor the female subgroup (Table 2). When we investigated an additional population of female Italians for the same SNPs, this analysis did neither show evidence of association (Table 3). In view of the very similar FOXO1 SNP frequencies and linkage dis- equilibrium (LD) structures in Germans and Italians (Table 5 and

Discussion

In the present study, we performed comprehensive haplotype tagging of the FOXO gene family members FOXO1, FOXO4, and FOXO6 in an extensive collection of LLI and younger controls, comprising altogether 2476 German individuals. This is the first investigation to analyze a possible association between FOXO4 or FOXO6, respectively, and human longevity, and the largest