Mechanisms of Ageing and Development 126 (2005) 333–339

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genes associated with ageing and life expectancy in the Jerusalem longitudinal study

Jochanan Stessman , Yoram Maaravi , Robert Hammerman-Rozenberg , Aaron Cohen , c c c b,c,* Lubov Nemanov , Inga Gritsenko , Nelly Gruberman , Richard P. Ebstein

aDepartment of Geriatric Rehabilitation, Hadassah Hospital, Hebrew University Medical Center, Mt. Scopus, Jerusalem bDepartment of Psychology, Scheinfeld Center of Human Genetics for the Social Sciences, Mount Scopus, Hebrew University, Jerusalem 91905, Israel

Available online 22 October 2004

Abstract

In an exploratory study, 11 common polymorphisms were examined for contributing to longevity including: apolipoprotein E (apoE), methylenetetrahydrofolate reductase (MTHFR), cathepsin D (CAD), superoxide dismutase 2 (SOD2), angiotensinogen (AGT) and insulin- like growth factor 2 (IGF2), Leiden factor 7, p53 oncogene, dopamine D4 receptor (DRD4) and the serotonin transporter (SERT). Genotype and allele frequencies of these genes were compared in 224 older (75 years) Jewish Jerusalem residents of Ashkenazi ethnicity to a group of 441 younger subjects (22 years). Nominally significant results provide suggestive evidence in the Ashkenazi group that apoE, MHTFR, SOD2, IGF2 ApaI, and factor VII are risk factors for a single outcome, survival to 75. Overall, the more genetically homogenous Ashkenazi ethnic group showed evidence for association in five genes examined suggesting that future studies in this population would gainfully focus on this ethnic group.

Keywords: Longevity; Polymorphisms; IGF2; Methylenetetrahydrofolate reductase; Apolipoprotein E; Superoxide dismutase; Angiotensinogen; D; Leiden factors V and VII; Dopamine D4 receptor; Serotonin transporter

1. Introduction

contributing to human longevity is only now slowly being elucidated.

1.1. Genes and aging in humans

Twin and adoption studies confirm a significant genetic component to human longevity (Carmelli, 1982). An oft- quoted Danish twin study (Herskind et al., 1996) estimated the heritabilty of life span to be 0.23 for males and 0.20 for females but a more recent Swedish study attributes perhaps as much as 50% of the variation in susceptibility to mortality to genetic factors (Yashin et al., 1999). Most researchers agree that heritability of lifespan in human is modest, <50%. Moreover, the role of specific genes in

The current case-control genetic study is based on the ongoing Jerusalem Longitudinal Cohort Study (JLCS) of 70- year-old (born in 1920/21) Jewish residents of Jerusalem initiated in 1990 (Bursztyn et al., 1999; Bursztyn et al., 1996; Cohen et al., 1995; Dresner-Pollak et al., 1996; Eliakim et al., 1996; Ginsberg et al., 1999; Maaravi et al., 1996; Stessman et al., 1995; Stessman et al., 1996; Stessman et al., 2000). The younger cohort was recruited from a non- clinical population in ongoing studies of normal personality (Ebstein et al., 2000; Zohar et al., 2003) and eating behavior (Bachner-Melman et al., 2004).

* Corresponding author. Tel.: +

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E-mail address: ebstein@mscc.huji.ac.il (R.P. Ebstein).

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1.3. Candidate genes studied

2.3. Genotyping

The SERT promoter region polymorphism was char- acterized as previously described by our laboratory (Ebstein et al., 1997). PCR amplification was carried out using the primers: SIL 1 50-ggC gTT gCC gCT CTg AAT gC-30 and SIL 2 50-gAg ggA CTg AgC Tgg AC A ACC AC-30. The reaction mixture was electrophoresed on a 2% metaphor gel (FMC) with ethidium bromide to screen for genotypes. For the VNTR, target DNA was amplified by PCR using the specific oligonucleotide primers: 50-GTCAGTATCA- CAGGCTGCGAG-30 and 50-TGTTCCTAGTCTTACGC- CAGTG-30 according to standard procedures (Ogilvie et al., 1996). The primer pair amplifies the region of intron 2 containing the 16–17 bp repetitive element.

frequency for a number of genes that we identified as worthwhile candidates that make biological sense for contributing to human lifespan: apolipoprotein E (Blanche et al., 2001; Rea et al., 2001; Slooter et al., 2001; Smith, 2002; Wang et al., 2001a), cathepsin D (Bi et al., 2003; Payton et al., 2003), insulin-like growth factor 2 (O’Dell et al., 1997, 1999; O’Dell and Day, 1998; Offen et al., 2001), superoxide dismutase 2 (De Benedictis et al., 1998; Mitrunen et al., 2001; Wang et al., 2001b), methylene tetrahydrofolate reductase (Kohara et al., 2003; Mattson et al., 2002; Visscher et al., 2003; Xia et al., 2003; Zuliani et al., 2002), Leiden factor V and VII (Hessner et al., 2001), angiotensinogen (Tan et al., 2001) and p53 oncogene (Bonafe et al., 1999). Additionally, we examined two ‘lifestyle’ genes that we also hypothesized might also contribute to lifespan, the dopamine D4 receptor linked in some studies to novelty or sensation seeking and the serotonin transporter linked in some studies to anxiety and depression (Ebstein et al., 2000).

follows. PCR amplification was carried out using a Reddy Mix kit (AB gene, Surrey, UK). The primers employed were forward 50-CTT CCTACC CTG CCC GCT CAT GCT GCT GCT CTA CTGG-30, and reverse 50-ACC ACC ACC GGC AGG ACC CTC ATG GCC TTG CGC TC-30. The reaction mixture contained the following components: 0.5 mM primers, 20 ng DNA and 10% DMSO in a total volume of 10 ml. After an initial denaturation step of 94 8C for 5 min, amplification was carried out for 35 cycles (948C for 30 s, 55 8C for 30 s and 72 8C for 90 s) in a Perkin-Elmer Cetus 9600 thermal cycler. A 5 min final extension at 72 8C was employed. The reaction mixture was electrophoresed on a 3% agarose gel (Ameresco: Ohio, USA) with ethidium bromide to screen for genotypes.

2. Materials and methods

2.1. Older subjects

Stessman and his colleagues have surveyed the social and medical conditions of this heterogeneous elderly population