Differences in the Association of Apolipoprotein E and Angiotensin-Converting Enzyme Gene Polymorphisms With Healthy Aging and Longevity: A Population-Based Study From Southern Italy

1 1,2 1 3 Davide Seripa, Marilisa Franceschi, Maria G. Matera, Francesco Panza, 4 1 1,2 3 Patrick G. Kehoe, Carolina Gravina, Giuseppe Orsitto, Vincenzo Solfrizzi, 5,6 7,8 1,2 Giovanni Di Minno, Bruno Dallapiccola, and Alberto Pilotto

1Laboratory of Geriatrics and Gerontology, Research Department, and 2Geriatric Unit, Medical Sciences Department, IRCCS

3Department of Geriatrics, Center for Aging Brain, Memory Unit, University of Bari, Italy. 4Care of the Elderly, Department of Clinical Science at North Bristol, University of Bristol, United Kingdom. 5Laboratory of Hemostasis and Thrombosis, Research Department, IRCCS ‘‘Casa Sollievo della Sofferenza,’’

6Department of Clinical and Experimental Medicine, ‘‘Federico II’’ University, Naples, Italy. Department, IRCCS ‘‘Casa Sollievo della Sofferenza,’’ San Giovanni Rotondo (FG), Italy. 8CSS-Mendel Institute, Rome, Italy.

We investigated the association of sex and age with the occurrence of apolipoprotein E (apoE) and angiotensin-converting enzyme (ACE) genotypes in healthy aging and longevity in 1344 healthy individuals and 64 centenarians. As compared to participants younger than 60 years, a significant higher frequency of the apoE/e2 was observed in men aged 60–90 years (p , .001) and in centenarians (p , .001). Logistic regression analysis confirmed this outcome in both participants aged 60–90 years (odds ratio [OR] ¼ 1.897; 95% confidence interval [CI], 1.227– 2.931) and centenarians (OR ¼ 3.263; 95% CI, 1.860–5.722). A further significant association of ACE/D allele and age was observed in centenarians (OR ¼ 2.135; 95% CI, 1.253–3.636). Heterosis was also observed at the ACE locus. No relationship between apoE and ACE polymorphism was found. These findings suggest a role of sex in the association of apoE and ACE gene polymorphisms with healthy aging and longevity.

longevity (3,4,6). While sex-specific differences between both of these polymorphisms and age-related diseases have been reported (13,14), their role in healthy aging and longevity in Caucasians is not well defined. The aim of this study was to evaluate the association of sex and age with the frequency of apoE and ACE polymorphisms, and in particular the frequency of apoE/e2 and ACE/D alleles in a population-based sample of healthy persons and in a group of centenarians living in southern Italy.

studies have indicated that longevity and the susceptibility to age-related diseases may be associated with different relative frequencies of common gene poly- morphisms (1,2), such as apolipoprotein E (apoE) and angiotensin-converting enzyme (ACE) (3–7).

single nucleotide polymorphisms in exon 4 of the apoE gene forming three common alleles: e2, e3, and e4, which encode for the three protein isoforms, E2, E3, and E4, and six common genotypes (8). These apoE polymorphisms are associated with several age-related diseases such as Alzheimer’s disease and vascular diseases (9), and seem to influence life expectancy (10). Thus, apoE polymorphism could be a marker for longevity and healthy aging (3,5).

M M ATERIALS AND ETHODS

Study Samples

A total of 1710 individuals (757 men and 953 women, mean age 41.31 6 13.63 years, range 18–90 years) were consecutively recruited among the personnel and relatives of the inpatients of the Istituto di Ricovero e Cura a Carattere Scientifico ‘‘Casa Sollievo della Sofferenza,’’ San Giovanni Rotondo, Italy, as part of a research program investigating

ACE polymorphism involves the insertion of a 287-base- pair Alu element in intron 16 of the gene. This insertion generates allele I (Aluþ), compared with the normal sequence defined as allele D (Alu(cid:1)) (11). Although less investigated than apoE, ACE polymorphism has also been associated with several age-related diseases (12), as well as

Digestion of PCR product with 4 U of HhaI (New England Biolabs, Inc., Beverly, MA) at 378C overnight produced a typical fragment pattern on a 4% agarose gel (20), one for each of the six apoE genotypes.

population genetics in Southern Italy. Only persons without a clinical history or evidence of neoplasms, infectious or inflammatory diseases, and/or acute disorders were initially considered for inclusion. After written informed consent was obtained, participants received a structured interview to evaluate the presence of age-related risk factors for degenerative diseases. Two hundred fifty-two individuals who had clinical evidence of vascular diseases (n ¼ 215, hypertension, ischemic cardiac, and cerebrovascular dis- eases, peripheral vascular diseases, diabetes) or cognitive impairment (n ¼ 37, Mini-Mental State Examination score lower than 26/30 points; 15) were excluded from the study. Moreover, 114 persons with serum levels of total choles- terol .220 mg/dL and/or low-density lipoprotein choles- terol .130 mg/dL and/or triglycerides .200 mg/dL (indicative of altered lipoprotein metabolism and increased risk of vascular diseases) were also excluded. Thus, a total of 1344 unrelated healthy Caucasians (79% of the overall population, 592 men and 752 women, mean age 41.31 6 13.63 years, range 22–90 years) were included in the study. All participants were Caucasians originating from Apulia, southern Italy.