The rs1333049 polymorphism located on locus 9p21.3 and extreme longevity in Spanish and Japanese cohorts. Toms Pins and Noriyuki Fuku contributed equally. Nobuyoshi Hirose and Alejandro Lucia share senior authorship. Received: 22 April 2013 / Accepted: 10 October 2013 # American Aging Association 2013

Abstract:
The rs1333049 (G/C) polymorphism located on chromosome 9p21.3 is a candidate to influence extreme longevity owing to its association with age-related diseases, notably coronary artery disease (CAD). We compared allele/genotype distributions of rs1333049 in cases (centenarians) and controls (younger adults, without (healthy) or with CAD) in two independent cohorts: Spanish (centenarians: n=152, 128 women, age 100-111 years; healthy controls: n=343, 212 women, age <50 years; CAD controls: n=98, 32 women, age 65 years) and Japanese (centenarians: n=742, 623 women, age 100-115 years; healthy controls: n=920, 511 women, age <60 years; CAD controls: n=395, 45 women, age 65 years). The frequency of the risk C-allele tended to be lower in Spanish centenarians (47.0%) than in their healthy (52.9%, p=0.088) or CAD controls (55.1%, p=0.078), and significant differences were found in genotype distributions (p=0.034 and p=0.045), with a higher frequency of the GG genotype in cases than in both healthy and CAD controls as well as a lower proportion of the CG genotype compared with healthy controls. In the Japanese cohort, the main finding was that the frequency of the C-allele did not differ between centenarians (46.4%) and healthy controls (47.3%, p=0.602), but it was significantly lower in the former than in CAD controls (57.2%, p<0.001). Although more research is needed, the present and recent pioneer findings (Rejuvenation Res 13:23-26, 2010) suggest that the rs1333049 polymorphism could be among the genetic contributors to exceptional longevity in Southern European populations, albeit this association does not exist in the healthy (CAD-free) Japanese population.

The single nucleotide polymorphism (SNP) rs1333049 (G/C) located on chromosome 9p21.3, a locus outside of annotated genes that confers susceptibility to CAD and myocardial infarction as shown with recent genome-wide scanning (McPherson 2010; Palomaki et al. 2010; Samani et al. 2007; Wellcome Trust Case Control Consortium 2007). This increased risk is independent of traditional risk factors including gender, age, obesity, smoking, hypertension, and hyperlipidemia (Schunkert et al. 2008). The C-allele (or the CC genotype) of rs1333049 has been associated with the risk or progression of CAD (Angelakopoulou et al. 2012; Bressler et al. 2010; Dandona et al. 2010; Ellis et al. 2010; Karvanen et al. 2009; McPherson 2010; Mendonca et al. 2011; Muendlein et al. 2009; O'Donnell et al. 2011; Palomaki et al. 2010; Plichart et al. 2012; Samani et al. 2009; Samani et al. 2007; Schunkert et al. 2008; Wellcome Trust Case Control Consortium 2007), and related disease phenotypes such as myocardial infarction (Buysschaert et al. 2010; Helgadottir et al. 2007; Koch et al. 2011; Scheffold et al. 2011; Shen et al. 2008b), atherosclerosis progression (Ye et al. 2008), peripheral arterial disease (Cluett et al. 2009), atherothrombotic manifestations (Arlestig and Rantapaa-Dahlqvist 2012), abdominal aortic aneurysm (Biros et al. 2010; Bown et al. 2008; Helgadottir et al. 2008), ischemic stroke (Karvanen et al. 2009; Smith et al. 2009), vascular dementia (besides Alzheimer's disease) (Emanuele et al. 2011) or type 2 diabetes (During the years 2008 to 2010, controls with CAD (n=99) were included in the study. Inclusion criteria for this group were (1) men or women aged 65 years and (2) diagnosed with CAD. We extracted genomic DNA from the saliva samples of patients (66 men, 33 women) in the cardiology department of two Spanish hospitals (Vall d'Hebron, Barcelona and Gregorio Maran, Madrid) during the summer of 2013. Written consent was obtained from each participant in the Japanese cohort. The study protocol was approved by the corresponding institutional ethics committee (National Institute of Health and Nutrition and Medical Research Institute, Tokyo Medical Dental University, Japan, for controls and Keio University, Japan, for centenarians) and was in accordance with the Declaration of Helsinki for Human Research of 1974 (last modified in 2008). All study participants were of the same Asian (Japanese) descent.

In the Spanish cohort, written consent was obtained from each participant. The study protocol was approved by the institutional ethics committee (Universidad Europea de Madrid, Spain) and was in accordance with the Declaration of Helsinki for Human Research of 1974 (last modified in 2008). All study participants were of the same Caucasian (Spanish) descent for three or more generations; the majority (~90%) of them were born and lived most of their lives in the same areas of Spain (Meseta Castellana, ~600 m altitude).

The Japanese cohort of cases was gathered from two prospective cohorts: the Tokyo Centenarians Study (TCS) and the Semi-Supercentenarians Study in Japan (SSC-J). A detailed description of population-based recruitments for the TCS has been previously reported (Gondo et al.). Cases (centenarians, n=175) were obtained from saliva samples during 2009 to 2012. The SSC-J is a nationwide longitudinal survey mainly consisting of individuals aged 105 years or older, which started in 2002. In 2002, we recruited 135 SSC in whole Japan. After 2002, our recruitment strategy has relied on responses to local governments and nursing homes in the whole country and direct inquiries to our research team. Consequently, a total of 450 centenarians (58 men, 392 women) were enrolled into SSC-J by the end of November 2011.