al. (2012) investigated the association of TERT SNPs with longevity in the general Danish population, but did not find any significant association. In this study, we investigated the association of the functional variable number of tandem repeat (VNTR) MNS16A of TERT with longevity in a population from Central Italy. MNS16A genotypes were determined for 1072 unrelated healthy individuals from Central Italy aged between 18 and 106 years old, divided into three gender-specific age classes based on demographic information and accounting for different survival rates between men and women. The first class consisted of individuals below 66 years old (below 73 years old for women), the second class consisted of individuals between 66 and 88 years old (between 73 and 91 years old for women), and the third class consisted of individuals above 88 years old (above 91 years old for women). TL was assessed using genomic DNA from whole blood of 72 selected individuals by a multiplex real-time PCR assay. The results showed that MNS16A is associated with longevity, with significant associations in Comparison 2 (Age Class 3 vs. Age Class 2) under both additive (odds ratio [O.R.] 0.749; p = 0.019) and dominant (O.R. 0.579; p = 0.011) models. The MNS16A*L allele is significantly underrepresented in Age Class 3 (O.R. 0.759; p = 0.020) compared to Age Class 2. A significant telomere attrition is reported along the three age classes (p = 0.0001), which remains significant only in L*/L* genotype carriers (p = 0.002) when the analysis was conducted according to MNS16A genotype. The TERT MNS16A*L allele appears negatively associated with longevity, and the concomitant significant telomere cross-sectional attrition rate observed for L*/L* genotype suggests that this polymorphism could influence human longevity by affecting TL. The study was approved by the Joint Ethical Committee (JEC) University of Camerino-Azienda ASUR Marche ZT-10 Camerino, in accordance with the Declaration of Helsinki in its revised edition and with international and local regulatory requirements.text population, to investigate the association of TERT VNTR MNS16A with telomere length. Genomic DNA was extracted from peripheral blood using a standardized salting-out method, and stored at -20°C until gene analysis. TERT VNTR MNS16A was genotyped using an allele-specific PCR method with a primer set and PCR Master Mix (2) (Fermentas). The amplification procedure consisted of an initial denaturing step at 95°C for 5 minutes followed by 35 cycles at 95°C for 30 seconds, 60°C for 45 seconds, and 72°C for 1 minute, with a final extension step at 72°C for 10 minutes. Each sample was genotyped in duplicate. The PCR products were visualized on a 2% agarose gel containing 0.25 mg/ml of ethidium bromide, and genotypes were assigned as L*/L*, S*/L* and S*/S*. 

In this study, we aimed to investigate the association of TERT gene with human longevity by analyzing the functional variable number of tandem repeat (VNTR) MNS16A. We conducted a cross-sectional study on a population from Central Italy, comprising of Caucasian individuals divided into three gender-specific age classes. A total of 1072 unrelated individuals aged between 18-106 years were recruited from the Marche region of Central Italy. The subjects provided information regarding their health condition to ensure that no pathological condition existed. The population was divided into three sex-specific age classes: for men [women], the first class consists of [missing information]; the second class consists of [missing information]; and the third class consists of [missing information]. 

To investigate the association of TERT VNTR MNS16A with telomere length, we selected a total of 72 samples (age-, sex- and genotype-matched) from the population. LTL was measured using monochrome multiplex Real-Time Quantitative PCR, which measures the relative amount of telomeric DNA (T) as compared to the amount of a single copy gene DNA (S) in the same sample (T/S ratio). The MNS16A genotypes were distributed as L*/L*, S*/L* and S*/S* and their distribution along the three age classes did not change significantly. However, significant associations of TERT VNTR MNS16A were found in Comparison 2 (Age Class 3 vs. Age Class 2) under both additive and dominant models. The MNS16A*L allele was significantly underrepresented in Age Class 3 compared to Age Class 2.in human longevity has been suggested by previous studies (Atzmon et al., 2010; Bernardes de Jesus et al., 2012). In this study, we aimed to investigate the effect of MNS16A genotype on telomere length (LTL) and its association with human longevity.

We analyzed a population and expressed the results as a T/S ratio. The mean T/S ratio was 1.111 ± 0.290, ranging from 0.683 to 1.833, and it followed a normal distribution in the study population (Kolmogorov-Smirnov test p = 0.153). We did not notice a significant difference between the sexes (females, 1.099 ± 0.283; males 1.124 ± 0.301; p = 0.721). As expected, LTL showed a significant age-dependent cross-sectional decrease (Age Class 1, mean T/S = 1.315 ± 0.298; Age Class 2, mean T/S = 1.029 ± 0.256; Age Class 3, mean T/S = 1.011 ± 0.223; p = 0.0001) (Fig. 2). LTL did not differ among the three MNS16A genotype groups (Table 2). However, the LTL attrition along the three age classes was significant only for MNS16A L*/L* genotype carriers (p = 0.002) (Table 2).