The minor allele frequencies of each SNP tested in MToR, RPToR, RicToR, and RPS6Ka1 in cases and controls are presented in Tables 2–5, together with results of chi-square analyses. Differences that appeared statisti- cally significant for two SNPs in RPToR (rs7225574 and rs2589143) and one SNP in RicToR (rs1239256) were lost after correction for multiple comparisons.

Phenotype–Genotype analyses

Phenotype–genotype comparisons for each SNP with 40 different aging-related phenotypes in cases showed weak associations for 5 of 6 SNPs in MToR, 38 of 61 in RPToR, and 5 of 7 in RicToR prior to correction for multiple test- ing (Supplementary Table S2). In all, 83 genotypic associa- tions were observed with blood pressure (BP) and related clinical phenotypes in the case of 23 of the RPToR SNPs. To complement the case–control analysis, multivariate tests for association of quantitative phenotypes with SNP allele counts (0, 1, and 2) were conducted for phenotypes deter- mined at Exam 1 and Exam 4 using canonical correlation. For BP, the mean of Exam 1 and Exam 2 data (substituting Exam 3 BP values if Exam 2 values were missing) were used. Variables were transformed when necessary to reduce skewness of the residuals from the regression of each phe- notype on age and case–control status. Wilk’s lambda test for association between the Exam 1 variables—log values of systolic and diastolic BP (log(SBP) and log(DBP)), serum cholesterol, body mass index, and log of subscap- ular skinfold thickness, adjusted for age at Exam 1—and case–control status was not significant (p = .29). Similarly, for the Exam 4 variables—grip strength, waist:hip ratio, body mass index, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, fasting serum glucose, log fasting insulin, log triglyceride level, and log of (101 − CASI [cognitive abilities screening instrument] score), adjusted for age at Exam 4 and case–control status—Wilk’s test was not significant (p = .22). Although we cannot rule out associations between particular age-related phenotypes and individual SNPs, the findings suggest that, overall, the actual number of associations observed did not exceed the expected numbers of associations that might be observed by chance alone.

Statistical analyses

Genotype frequencies of SNPs were evaluated for devia- tion from Hardy–Weinberg equilibrium. The Pearson chi- square test was used to compare cases and controls for difference in genotype frequencies. The 95% confidence intervals for allele frequencies were calculated as described (32). For analyses of phenotypes, linear regression models were used for continuous variables (with log transforma- tion of skewed variables) and logistic regression models were used for categorical variables, to test for associations with genotypes, adjusting for case–control status and age at baseline. The analyses used the SAS statistical package

Results

characteristics of Participants

of the HHP and HAAS study population at the 1991–1993 examination when blood was drawn for genotyping in the present study (Exam 4 of the cohort). The more limited set of parameters recorded at Exam 1 (1965–1968) are also shown. Unadjusted data are shown in Supplementary Table S1. Presented are biological characteristics, general health status, disease prevalence, and functional status of long-lived “cases” (longevity phenotype) and average-lived “controls” (average phenotype). The long-lived cases were older, leaner (based on lower waist:hip ratio), had lower plasma glucose level, lower plasma insulin level, better self-rated health, and lower prevalence of cardiovascular disease (hypertension, coronary heart disease, and stroke) and cancer. The cases appeared better able to walk 0.8 km than controls, but grip strength was similar. Cognitive score was slightly higher in cases. When adjusting for age, the differences between long-lived cases became even starker,

Discussion

The present nested case–control study found no associa- tion of genetic variation in mTOR pathway genes MToR, RPToR, RicToR, and RPS6Ka1 with longevity (survival to age 95 years or older) in American men of Japanese

Table 1. Age-Adjusted Baseline Characteristics of Case and Control Participants (mean ± SD)