Study participants

The study sample comprised 1447 LLI with an age range of 95–110 years (mean age: 98.8 years) and 1029 younger con- trols (60–75 years; mean age: 66.8 years). About 75% of the LLI were women. The controls were similar to the LLI in terms of gender (75% women), ancestry, and geographic origin. There are no mortality data available for the controls. However, based on current predictions, only 1.5% of all 60-year-old and 1.8% of all 75-year-old German women will become 100 years. The study participants provided, as part of a health and family history questionnaire, personal and medical information. A detailed description of the study population and recruitment procedures is provided elsewhere (Nebel et al., 2005). All subjects gave written, informed consent prior to enrolment. The study was approved by the Ethics Committee of the University Hospital Schleswig-Holstein in Kiel.

Table 5 Comparison of minor allele frequencies of FOXO1 single-nucleotide polymorphisms (SNPs) in German, Italian, Chinese, and Japanese controls

The Italian women sample consisted of 166 LLI (90–109 years, mean age: 98.39 years) and 216 younger controls (18–48 years; mean age: 31.06 years) obtained from the Southern Italian Cen- tenarian Study (SICS). SICS LLI were thoroughly investigated for demographic and clinical characteristics and they were enrolled by Associazione Longevita` (Anselmi et al., 2009). All subjects gave written informed consent to the study. The study was approved by the Institutional Review Board of the ‘Istituto di Ricovero e Cura a Carattere Scientifico’, Multimedica, Milano, Italy.

minAF minAF minAF minAF controls controls controls controls Germans Italians Chinese† Japanese(cid:2) rs2755209 0.37 0.40 0.29 0.33§ rs2755213 0.09 0.11 0.42 NA rs17630266 0.03 0.02– 0.37 0.39††

Associated SNPs from Li et al. (

) are in bold. mi

F, minor allele frequency; NA, no information available. †Data from Li et al. (

). (cid:

)Data from Kojima et al. (

). §Based on SNP rs

from Kojima et al. (

) that is in strong linkage disequilibrium (LD) with rs

(r

=

, using JPT Ha

ap data). –Based on imputed data. ††Based on SNP rs

from Kojima et al. (

) that is in strong LD

Genotyping

Genotyping of the German samples was performed using the SNPlexTM Genotyping System and TaqMan(cid:2) SNP Genotyping Assays (both Applied Biosystems, Foster City, CA, USA) on an automated platform (Hampe et al., 2001). For quality control purposes, positive controls on each genotyping plate were checked for consistency. All SNPs were tested for deviation from the HWE. The call rate for each marker exceeded 95.8%.

††Based on SNP rs2297626 from Kojima et al. (2004) that is in strong with rs17630266 (r2 = 0.95, using JPT HapMap data).

Further studies are needed to clarify in more detail the in vivo roles of the FOXO genes, and in particular of FOXO3A, in human longevity.

(A) (B) (C) (D) 6 6 6 6 9 3 6 9 3 6 9 3 6 9 3 6 0 1 2 0 1 2 0 1 2 0 1 2 2 2 0 2 2 0 2 2 0 2 2 0 5 5 3 5 5 3 5 5 3 5 5 3 5 5 6 5 5 6 5 5 6 5 5 6 7 7 7 7 7 7 7 7 7 7 7 7 2 2 1 2 2 1 2 2 1 2 2 1 s s s s s s s s s s s s r r r r r r r r r r r r 15 14 18 13 53 34 86 52 5 2 18 45

Fig. 2 LD structure of FOXO1 in Germans (A), Italians (B), Chinese (C), and Japanese (D). The depicted single-nucleotide polymorphisms were used in the study of Chinese (Li et al., 2009). The linkage disequilibrium plots are based on the measure r2 and were generated with Haploview using HapMap data of Han Chinese (CHB) and Japanese (JPT) as well as data of German and Italian control individuals from this study.

References

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