The phenotypic characteristics were adjusted for age and gender when controls were compared with offspring and for gender when controls were compared with centenarians. All values are mean (SE).

sequence variants in the gene, among which 5 were previously unknown unique variants that were not reported in various SNP databases (Table 3). The locations of these variants are shown in Fig. 4. Nine variants in the hTERT gene are in the coding region, including two nonsynonymous variants, 893 G > A (Ala-279 Thr) and 3242 G > A (Ala-1062 Thr), which are rare and found only in controls (Table 3). In silico analysis using SIFT (sorts intolerant from tolerant substitutions) (30) and PolyPhen (31), which predict the effects of amino acid changes on protein function, indicates that these changes are likely to be benign. Interestingly, rare synonymous or intronic variants in hTERT are enriched in centenarians (n = 19) compared with controls (n = 3) (Table 2; P = 0.041). In addition, three previously un- described intronic variants in hTERC were also found, two of which were rare and found only in centenarians (Table 3).

Telomere Length, Age-Associated Diseases, and Lipid Profiles of

Healthy Aging. Association analysis between telomere and major age-related diseases among centenarians, their fi spring, and controls indicated that signi cantly shorter mere lengths (adjusted for age, gender, and group) are P in subjects with hypertension ( = 0.006), the metabolic P P drome ( = 0.03), or diabetes ( = 0.03) compared with without these disorders (Fig. 2). Moreover, when we tested relation between telomere length and cognitive function centenarians, centenarians with impaired cognitive [Mini-Mental State Examination (MMSE) score ≤ 25] fi signi cantly shorter telomeres as compared with with normal cognitive function after adjustment for age gender (Fig. 2; P = 0.02). We then tested whether fi length correlated with the lipid pro les, which are known dictors of aging-related diseases such as coronary artery and metabolic syndromes (29). These analyses revealed telomere length is associated with the lipid profi les of aging, showing a positive association with increased sizes of LDL (P = 0.0001) and HDL (P = 0.03), percentage large particle sizes of LDL (P = 0.0002) and HDL (P = and levels of Apo-A1 (P = 0.005) and HDL (P = 0.04) (Table whereas there is a negative association with very LDL (P = 0.008). Total cholesterol, triglycerides, LDL, and levels show no signifi cant correlation (Table 2). These suggest that longer telomeres are positively correlated with lipid profi les of healthy aging.

We genotyped the four common hTERT variants, with a minor allelefrequency (MAF)of> 5%[IVS1– 187T> C,973G> A(Ala- > > 305 Ala), 3097 C T (His-1013 His), IVS16+99 C T], in 73 centenarians and 49 controls. Information on telomere length was fi available for all these individuals. We did not nd an association of the IVS1– 187 T > C, 973 G > A (Ala-305 Ala), and IVS16+99 C > fi T with longevity. However, signi cant associations were detected with the 3097 C > T (His-1013 His) variant: the T allele is sig- nifi cantly enriched in centenarians (21.8%) compared with con- trols (8.2%; P = 0.026) (Table 3), and both CT and TT genotypes fi are signi cantly enriched in centenarians compared with controls (26.8%and8.5%vs.12.2%and2%,respectively;P =0.035)(Table 4). In contrast, no signifi cant association between the hTERC > common variant (IVS+63 T C) and longevity was found (Table 4). Haplotype analysis [see SI Fig. S1 for a linkage disequilibrium (LD)plot derived from the four commonvariants] revealedthat of fi the four most common haplotypes, Hap 1 is signi cantly depleted in centenarians compared with controls (34.9% vs. 53.1%; P = 0.0056), whereas Hap 3 is signifi cantly enriched in centenarians compared with controls (13.7% vs. 8.2%; P = 0.007) when adjusted for age and gender (Table 5). Interestingly, a rare haplotype in fi controls, Hap 6, is found signi cantly more frequently in cen- P tenarians (1% vs. 6.8%; = 0.024) (Table 5). Association analysis

Variation in the Telomerase Genes, Longevity, and Telomere Length.

We performed a comprehensive sequence analysis of hTERT and hTERC genes to detect all possible genetic variants, in- cluding the rare variants that may be enriched in centenarians, – fl throughout the coding exons and exon intron anking regions in n n centenarians ( = 100) and controls ( = 80) using 2D gene scanning and DNA sequencing (Fig. 3). We found a total of 15

TIS ratio  

Fig. 2. Telomere length (adjusted for age, gender, and group) according to the absence (gray bars) or presence (white bars) of various age-related diseases the study subjects. Number of individuals analyzed in each group: hypertension (n = 125), without hypertension (n = 142), with the metabolic syndrome (n 59), without the metabolic syndrome (n = 286), diabetes (n = 34), without diabetes (n = 276). The MMSE was performed in centenarians only: MMSE score (gray bars, n = 46) or >25 (white bars, n = 32). *P < 0.05.

netically controlled longevity traits. This approach has previously fi led to the identi cation of longevity phenotypes such as lip- oprotein sizes and the subsequent discovery of corresponding longevity genotypes (32– 34). In the present investigation, we employed the same study design to delineate the relation be- tween telomere lengths, longevity, and diseases of aging as well as to gain insights into the potential role(s) of genetic variations in the hTERT and hTERC genes on these phenotypes.