Hostname: page-component-586b7cd67f-tf8b9 Total loading time: 0 Render date: 2024-11-26T12:19:29.016Z Has data issue: false hasContentIssue false

ApoE genotype, cardiovascular risk and responsiveness to dietary fat manipulation

Symposium on ‘Molecular basis for diseases’

Published online by Cambridge University Press:  30 April 2007

A. M. Minihane*
Affiliation:
Hugh Sinclair Unit of Human Nutrition, School of Chemistry, Food Biosciences and Pharmacy, University of Reading, Reading RG6 6AP, UK
L. Jofre-Monseny
Affiliation:
Institute of Human Nutrition and Food Science, Christian Albrechts University, Hermann-Rodewald-Strasse 6, 24098 Kiel, Germany
E. Olano-Martin
Affiliation:
Hugh Sinclair Unit of Human Nutrition, School of Chemistry, Food Biosciences and Pharmacy, University of Reading, Reading RG6 6AP, UK
G. Rimbach
Affiliation:
Institute of Human Nutrition and Food Science, Christian Albrechts University, Hermann-Rodewald-Strasse 6, 24098 Kiel, Germany
*
*Corresponding author: Dr Anne M. Minihane, fax +44 118 9310080, email [email protected]
Rights & Permissions [Opens in a new window]

Abstract

Cardiovascular risk is determined by the complex interactions between genetic and environmental factors. The apoE genotype represents the most-widely-studied single nucleotide polymorphism in relation to CVD risk, with >3600 publications cited in PubMed. Although originally described as a mediator of lipoprotein metabolism, the lipoprotein-independent functions of apoE are being increasingly recognised, with limited data available on the potential impact of genotype on these metabolic processes. Furthermore, although meta-analyses suggest that apoE4 carriers may have a 40–50% increased CVD risk, the associations reported in individual studies are highly heterogeneous and it is recognised that environmental factors such as smoking status and dietary fat composition influence genotype–phenotype associations. However, information is often derived from observational studies or small intervention trials in which retrospective genotyping of the cohort results in small group sizes in the rarer E2 and E4 subgroups. Either larger well-standardised intervention trials or smaller trials with prospective recruitment according to apoE genotype are needed to fully establish the impact of diet on genotype–CVD associations and to establish the potential of dietary strategies such as reduced total fat, saturated fat, or increased antioxidant intakes to counteract the increased CVD burden in apoE4 carriers.

Type
Research Article
Copyright
Copyright © The Author 2007

Abbreviations:
HDLC

HDL-cholesterol

LDLC

LDL-cholesterol

The impact of single nucleotide polymorphisms on risk of chronic diseases such as CVD, and the ability of dietary factors to manipulate genotype–phenotype associations, is being increasingly recognised. Undoubtedly, the most-widely-studied gene variant in relation to CVD is the apoE ε (ε2, ε3, ε4) genotype. Since its discovery in 1973 the central role of the apoE protein in lipoprotein metabolism has been comprehensively investigated and reported. The 40–50% higher risk of CVD in apoE4 carriers (Song et al. Reference Song, Stampfer and Liu2004) has been traditionally attributed to moderately higher circulating cholesterol and TAG levels. However, it is becoming increasingly recognised that an effect on lipoprotein metabolism alone cannot explain the disease differential and that the impact of an apoE4 genotype is largely lipoprotein independent. Roles of macrophage-derived apoE protein on vascular health and atherogenesis are being identified, with apoE thought to impact on oxidative status and in an autocrine and paracrine manner affect macrophage, vascular smooth muscle cell, endothelial cell and platelet function. An impact of genotype on these localised functions of apoE could in part explain the impact of genotype on CVD pathology, as will be discussed.

Additionally, apoE genotype has been shown to affect the responsiveness to the total fat content and fatty acid composition of the diet. Manipulation of dietary fat content may serve as a means of reducing the increased CVD burden associated with an apoE4 genotype.

ApoE structure and tissue sources

ApoE was first described as a component of VLDL in the circulation (Shore & Shore, Reference Shore and Shore1973). The full amino acid sequence was elucidated in 1982, with the mature 299 amino acid 34 kDa acid protein resulting from the proteolytic cleavage of the 317 amino acid product of the apoE gene (Rall et al. Reference Rall, Weisgraber and Mahley1982). ApoE is found in the circulation associated with chylomicrons, VLDL and HDL at a typical concentration of 20–60 mg/l (Bhatnagar & Durrington, Reference Bhatnagar and Durrington1993).

The protein assumes a typical apo form with two structural domains (Fig. 1; from Hatters et al. Reference Hatters, Peters-Libeu and Weisgraber2006). The amino terminal (22 kDa) comprises residues 1–191 and ‘houses’ the lysine- and arginine-rich receptor-binding region contained between amino acids 136 and 150 (Innerarity et al. Reference Innerarity, Friedlander, Rall, Weisgraber and Mahley1983). The carboxyl terminal (10 kDa) consists of residues 225–299 and contains the major lipid-binding determinants that anchor apoE to the lipoprotein (Wetterau et al. Reference Wetterau, Aggerbeck, Rall and Weisgraber1988). These domains are separated by a protease-sensitive hinge region (Wetterau et al. Reference Wetterau, Aggerbeck, Rall and Weisgraber1988). Despite the independent folding of the two domains, there are recognised domain interactions (Dong & Weisgraber, Reference Dong and Weisgraber1996).

Fig. 1. Key structural elements of apo E (reprinted from Hatters et al. Reference Hatters, Peters-Libeu and Weisgraber2006, with permission from Elsevier). (a) The amino-terminal domain consists of a four-helix bundle that contains the LDL receptor-binding region of the protein contained between amino acids 136–150 in helix 4. Contained within the ‘hinge region’, amino acid 172 is thought to be essential for receptor binding. The carboxyl-terminal contains the lipoprotein-binding region. (b) The model demonstrates the impact of the replacement of Cys with Arg on position 112 in the protein. This replacement facilitates the interaction between Arg 61 and Glu 255, which mediates closer contact between the amino-terminal and carboxyl-terminal domains.

The structure of the carboxyl-terminal domain is unknown but is predicted to be mostly α-helical (Nolte & Atkinson, Reference Nolte and Atkinson1992), whilst the three-dimensional structure of the amino-terminal domain in lipid-free solution has been determined by X-ray crystallographic studies to be an elongated globular four-helix bundle. Helix 1 pairs with helix 2, and helix 3 with helix 4, arranged in an anti-parallel mode, with the hydrophobic faces oriented towards the interior of the bundle (Wilson et al. Reference Wilson, Wardell, Weisgraber, Mahley and Agard1991). ApoE genotype impacts on the three-dimensional orientation of the apoE regions and amino-terminal–carboxyl-terminal interactions, which affect receptor binding and lipoprotein apoE distribution, as will be discussed (see pp. 185–186). For more detailed information on apoE structure and structure–function relationships, see Hatters et al. (Reference Hatters, Peters-Libeu and Weisgraber2006).

ApoE is synthesised mainly in the liver, with hepatocytes being the main producers. It has been estimated that between 20 and 40% of the total apoE protein is produced by extrahepatic tissues, with the brain and the monocyte-derived macrophages expressing relatively high amounts (Basu et al. Reference Basu, Ho, Brown, Bilheimer, Anderson and Goldstein1982; Kayden et al. Reference Kayden, Maschio and Traber1985; Newman et al. Reference Newman, Dawson, Rudel and Williams1985; Wang-Iverson et al. Reference Wang-Iverson, Gibson and Brown1985). ApoE is also synthesised by a range of other tissues, including steroidogenic organs such as the adrenal glands, testes and ovary (Blue et al. Reference Blue, Williams, Zucker, Khan and Blum1983; Polacek et al. Reference Polacek, Beckmann and Schreiber1992), lungs (Dawson et al. Reference Dawson, Lukaszewski, Ells, Malbon and Williams1989), kidney (Wallis et al. Reference Wallis, Rogne, Gill, Markham, Edge, Woods, Williamson and Humphries1983) and adipose tissue (Zechner et al. Reference Zechner, Moser, Newman, Fried and Breslow1991), and in the retinal pigment epithelial cells (Ishida et al. Reference Ishida, Bailey, Duncan, Chalkley, Burlingame, Kane and Schwartz2004).

Role of apoE in lipoprotein metabolism

ApoE is known to play a multi-functional role in lipoprotein metabolism, potentially acting as a cofactor in VLDL synthesis, the hydrolysis of VLDL remnants to produce LDL and as a high-affinity ligand for the receptor-mediated cellular removal of lipoprotein remnants. Although apoE is a constituent of Golgi VLDL, there are inconsistencies in the literature in relation to the essentiality of apoE in hepatic VLDL synthesis and secretion (Schaefer et al. Reference Schaefer, Gregg, Ghiselli, Forte, Ordovas, Zech and Brewer1986; Fazio & Yao, Reference Fazio and Yao1995; Huang et al. Reference Huang, Ji, Brecht, Rall, Taylor and Mahley1999). Undoubtedly, the most important role of apoE in lipoprotein metabolism is as a high-affinity ligand for receptors of the LDL receptor family, and the impact of genotype on lipoprotein metabolism is thought to be largely the result of an effect on the receptor binding activity of apoE. Members of this family include the LDL receptor, the LDL receptor-related protein, the VLDL receptor and the apoE receptor 2 (Strickland et al. Reference Strickland, Gonias and Argraves2002).

The apoE–receptor interactions, which mediate the cellular uptake of VLDL and chylomicron remnants, have been widely studied (Bradley & Gianturco, Reference Bradley and Gianturco1986; Mahley, Reference Mahley1988). It is thought that the basic amino acids located between residues 136 and 150, which produce a large region of positive electrostatic potential, are important for its interaction with the acidic amino acid ligand-binding region of members of the LDL receptor family (Weisgraber, Reference Weisgraber1994). Since single amino acid substitutions in this portion of the protein result in defective binding but not in complete abolition of binding activity, it is considered that the basic amino acids cooperate in the interaction with the receptor (Wilson et al. Reference Wilson, Wardell, Weisgraber, Mahley and Agard1991). Subtle changes around the LDL receptor-binding region also lead to defective receptor activity, as will be discussed.

ApoE receptor 2 (also termed LRP8) is structurally distinct from other family members in having a longer cytoplasmic domain. Furthermore, its pattern of tissue distribution is different from that of other receptors (Kim et al. Reference Kim, Iijima, Goto, Sakai, Ishii, Kim, Suzuki, Kondo, Saeki and Yamamoto1996), with apoE receptor 2 lacking in the liver but found abundantly in the brain and in several other tissues such as platelets and testes (Riddell et al. Reference Riddell, Vinogradov, Stannard, Chadwick and Owen1999). It is thought that apoE receptor 2 is involved in the role of apoE in cellular signalling pathways, which is at present poorly understood. Furthermore, the precise apoE sequence that binds to this receptor has not been established (Li et al. Reference Li, Kypreos, Zanni and Zannis2003).

ApoE also binds to scavenger receptor type BI and cell glycosaminoglycans, including heparin and heparin sulphate proteoglycans. ApoE binding to heparin sulphate proteoglycans is thought to be an initial step in the localisation of apoE-containing lipoproteins to the surface of different cell types. The best understood physiological role for this interaction is the hepatic clearance of remnant lipoproteins, contributing to the initial sequestration and subsequent uptake steps, either in association with LDL receptor-related protein or acting alone (Mahley & Ji, Reference Mahley and Ji1999; Libeu et al. Reference Libeu, Lund-Katz, Phillips, Wehrli, Hernaiz, Capila, Linhardt, Raffai, Newhouse, Zhou and Weisgraber2001).

Impact of apoE genotype on protein structure and function

In man the apoE gene is mapped to chromosome 19 in a cluster with apoC1 and apoC2. It extends for 3610 bases starting at 50 100 879 bp from pter to 50 104 489 bp from pter and consists of four exons (44, 66, 193 and 869 bp) and three introns (760, 1092 and 592 bp; Paik et al. Reference Paik, Chang, Reardon, Davies, Mahley and Taylor1985). Currently, forty-five single nucleotide polymorphisms have been identified for the apoE gene (National Center for Biotechnology Information (2006) single-nucleotide polymorphism database), twenty-seven in the intronic region and eighteen in coding regions (Table 1).

Table 1. Polymorphisms found in apoE gene exons (data from National Center for Biotechnology Information (2006) single-nucleotide polymorphism database)

SNP ID, single-nucleotide polymorphism identification; N/A, not available.

A common and widely characterised genotype is the apoE-ε missense mutations that result in three allelic isoforms ε2, ε3 and ε4 (Table 1). The protein products differ in the amino acid present at residue 112 (rs429358) and 158 (rs7412) of the protein (Tables 1 and 2). ApoE2 contains 112 Cys/158 Cys, apoE3 112 Cys/158 Arg, and apoE4 112 Arg/158 Arg (Weisgraber et al. Reference Weisgraber, Rall and Mahley1981; Rall et al. Reference Rall, Weisgraber and Mahley1982). Although the amino acids alterations do not occur within the receptor binding region (amino acids 136–150), the substitutions at positions 112 and 158 are known to impact on the salt bridge formation within the protein, which ultimately impacts on the receptor binding activity and lipoprotein ‘preference’ of the apoE protein. ApoE3 and apoE4 have comparable LDL receptors affinity, but the binding of apoE2 is 50–100 times weaker (Weisgraber et al. Reference Weisgraber, Innerarity and Mahley1982; Weisgraber, Reference Weisgraber1994). The replacement of an arginine residue with cysteine at position 158 is thought to eliminate a salt bridge between Asp154 and Arg 158 with a new bridge forming between Arg 150 and Asp 154, which dramatically alters the conformation of the receptor binding domain (Hatters et al. Reference Hatters, Peters-Libeu and Weisgraber2006; Fig. 1). The impact of genotype on the binding of apoE to other members of the LDL-receptor family is relatively unknown; although no substantial impact of isoform on LDL receptor-related protein- and VLDL receptor–apoE interactions has been observed in a series of in vitro binding studies (Ruiz et al. Reference Ruiz, Kouiavskaia, Migliorini, Robinson, Saenko, Gorlatova, Li, Lawrence, Hyman, Weisgraber and Strickland2005).

Table 2. ApoE isoform amino acid differences and physio-chemical changes

CM, chylomicrons.

The Cys112 to Arg112 substitution in apoE4, although not appearing to appreciably influence receptor binding, is thought to impact on both protein stability and carboxyl-terminal and amino-terminal domain interactions (for review, see Hatters et al. Reference Hatters, Peters-Libeu and Weisgraber2006). An arginine moiety at this position is thought to impact on the conformation of Arg61, allowing its interaction with an acidic Glu255 residue in the carboxyl-terminal (Fig. 1). This interaction affects the protein conformation, resulting in a ‘molten globule’ structure (Morrow et al. Reference Morrow, Hatters, Lu, Hochtl, Oberg, Rupp and Weisgraber2002) with a preference for larger VLDL and chylomicron remnants, in contrast to apoE2 and apoE3, which prefer smaller cholesterol-rich HDL particles. The higher lipid-binding affinity of apoE4 is not influenced by the particle size (Saito et al. Reference Saito, Dhanasekaran, Baldwin, Weisgraber, Phillips and Lund-Katz2003).

This impact on protein structure also affects molecular stability, with susceptibility of the isoforms to degradation being in the following order E4>E3>E2 (Acharya et al. Reference Acharya, Segall, Zaiou, Morrow, Weisgraber, Phillips, Lund-Katz and Snow2002).

ApoE allelic frequency and genotype distributions

Globally, the apoE allelic distribution shows substantial variation, with an allele frequency of 60–90% for the wild-type ε3 allele (Corbo & Scacchi, Reference Corbo and Scacchi1999; Singh et al. Reference Singh, Singh and Mastana2006).

The studies reviewed by Eichner et al. (Reference Eichner, Dunn, Perveen, Thompson, Stewart and Stroehla2002) demonstrate that approximately 65% of Caucasian populations are homozygous ε3/ε3, 19% are ε3/ε4, 10% are ε2/ε3, 4% are ε2/ε4, 2% are ε4/ε4 and 0·5–1% are ε2/ε2. In Europe there is a geographic cline, with 2-fold higher prevalence of the ε4 allele in northern Europe compared with southern Europe (Corbo & Scacchi, Reference Corbo and Scacchi1999; Eichner et al. Reference Eichner, Dunn, Perveen, Thompson, Stewart and Stroehla2002; Singh et al. Reference Singh, Singh and Mastana2006; Table 3), which is likely to make a contribution to the north–south differences in CVD incidence observed.

Table 3. ApoE allelic distribution in select populations worldwide (derived from Singh et al. Reference Singh, Singh and Mastana2006)

* Listed in order of ε4 allele.

European countries.

ApoE genotype and cardiovascular risk and incidence: impact of age and gender

Over the last three decades numerous studies using a variety of CHD end points, including clinically- and angiographically-defined CHD, have investigated the impact of apoE genotype on CHD risk. The main studies have been summarised in two meta-analyses (Wilson et al. Reference Wilson, Schaefer, Larson and Ordovas1996; Song et al. Reference Song, Stampfer and Liu2004). The Wilson et al. (Reference Wilson, Schaefer, Larson and Ordovas1996) analysis summarises data from fourteen published observational studies, with carriers of the ε4 allele having an overall OR for CHD of 1·26 (95% CI 1·13, 1·41) and a non-significant OR of 0·98 (95% CI 0·85, 1·14) evident in ε2 carriers. On removing the Utermann et al. (Reference Utermann, Hardewig and Zimmer1984) study, which demonstrated a cardio-protective effect of E4 and reported results that were clearly divergent from all other studies, an OR of 1·44 (95% CI 1·27, 1·62) was observed. This finding is in agreement with the more-recent meta-analysis (Song et al. Reference Song, Stampfer and Liu2004), which includes data from 15 492 CHD cases and 32 965 controls. Overall OR of 1·42 (95% CI 1·26, 1·61) and 0·98 (95% CI 0·66, 1·46) were observed for the E4 and E2 subgroups. However, findings from the forty-eight studies included are highly heterogeneous with mean OR values derived from the individual studies ranging from 0·68 to 4·35 in ε4 carriers compared with the wild-type E3/E3 genotype. Such heterogeneity is likely to be attributable to an array of factors, including environmental factors such as smoking status and background diet, and also the age and gender of the study cohort.

Currently, a comprehensive review of the impact of age and gender on apoE genotype–CHD associations is distinctly lacking. Data from the Framingham Offspring study (Wilson et al. Reference Wilson, Myers, Larson, Ordovas, Wolf and Schaefer1994; Lahoz et al. Reference Lahoz, Schaefer, Cupples, Wilson, Levy, Osgood, Parpos, Pedro-Botet, Daly and Ordovas2001; Elosua et al. Reference Elosua, Ordovas, Cupples, Fox, Polak, Wolf, D'Agostino and O'Donnell2004) suggest a protective effect of an E2 genotype and a greater sensitivity to the deleterious effects of an E4 genotype in females compared with males. In relation to age, it appears that the impact of genotype on CVD risk is attenuated with age (Jarvik et al. Reference Jarvik, Austin, Fabsitz, Auwerx, Reed, Christian and Deeb1994; Ilveskoski et al. Reference Ilveskoski, Perola, Lehtimaki, Laippala, Savolainen and Pajarinen1999), with a lack of association of genotype with disease risk in older cohorts (Kuusisto et al. Reference Kuusisto, Mykkanen, Kervinen, Kesaniemi and Laakso1995). For example, in the Helsinki Sudden Death Study (Ilveskoski et al. Reference Ilveskoski, Perola, Lehtimaki, Laippala, Savolainen and Pajarinen1999), which conducted lesion staining of the coronary arteries of 700 individuals, age×genotype interactions were observed, with an impact of genotype only present in the group who were <53 years old.

It is speculated that the apparent age-related weakening of the association may be (a) attributable to the masking effect of an overall ‘at-risk’ phenotype, which is reflected in more extensive atherogenesis, reducing the variability and the association with any one genetic factor, or (b) because individuals who are particularly sensitive to the genotype-mediated effects may have already died and are therefore not included in the analysis of older cohorts.

ApoE genotype and physiological determinants of risk for CVD

Traditionally, an increased CVD risk in E4 carriers has been attributable to higher circulating total cholesterol and LDL-cholesterol (LDLC) in E4 carriers. As will be discussed, the sometimes moderate and often non-significantly higher circulating cholesterol levels in E4 carriers are not likely to explain the 40–50% higher CVD risk observed. Furthermore, the retention of a significant impact of genotype when correction is made for recognised lipid risk markers of disease (Terry et al. Reference Terry, Howard, Mercuri, Bond and Crouse1996; Humphries et al. Reference Humphries, Talmud, Hawe, Bolla, Day and Miller2001; Lahoz et al. Reference Lahoz, Schaefer, Cupples, Wilson, Levy, Osgood, Parpos, Pedro-Botet, Daly and Ordovas2001) suggests that the effect is partly mediated by lipid-independent mechanisms.

ApoE genotype and blood lipid levels

ApoE genotype and LDL-cholesterol levels

It has been documented that apoE genotype accounts for 7% of the variance of total cholesterol in healthy Caucasian individuals (Davignon et al. Reference Davignon, Gregg and Sing1988), and it has been suggested that an adverse cholesterol profile in E4 carriers could largely explain the increased risk of coronary events in this subgroup.

In most of the populations studied, regardless of age and health status, the ε4 allele has been associated with higher LDLC and apoB concentrations relative to E2 carriers (Table 4). However, relative to E3/E3 carriers only moderate differences in cholesterol exist, with the differences often not significant. In the studies included in Table 4 LDLC concentrations for E4 and E2 carriers are on average 8·3% higher and 14·2% lower respectively than those for E3 homozygotes, with the cholesterol-lowering effect of the ε2 allele known to be greater than the cholesterol-raising effect of ε4 allele (Davignon et al. Reference Davignon, Gregg and Sing1988; Hallman et al. Reference Hallman, Boerwinkle, Saha, Sandholzer, Menzel, Csazar and Utermann1991; Schaefer et al. Reference Schaefer, Lamon-Fava, Johnson, Ordovas, Schaefer, Castelli and Wilson1994).

Table 4. The impact of apoE genotype on LDL-cholesterol levels (E2/E4 excluded if present)

E2, E2/E2+E2/E3; E3, E3/E3; E4, E3/E4+E4/E4; MI, myocardial infarction, FHBI, familial hyperbetaproteinaemia; HRT, hormone-replacement therapy; LPG, lipoprotein glomerulopathy; N/A, not available; approx, approximately; ↑, increase; ↓, decrease.

How does apoE genotype modulate LDL-cholesterol levels?

The lower plasma LDLC in E3/E2 and E2/E2 subjects has been attributed to a number of mechanisms, including increased hepatic receptor-mediated LDL removal, lower VLDL to LDL conversion rates and decreased intestinal cholesterol absorption.

In E2 carriers defective binding of the apoE2 protein to receptors will lead to reduced hepatic VLDL and chylomicron remnant uptake, resulting in a reduced hepatic cholesterol load, which in turn will trigger up-regulation of the LDL receptor (Gregg & Brewer, Reference Gregg and Brewer1988). Increased LDL receptor expression together with reduced receptor affinity of the apoE protein would be predicted to increase apoB100-mediated LDL removal by the LDL receptor (Howard et al. Reference Howard, Gidding and Liu1998). In a number of human biokinetic studies a higher fractional catabolic rate of LDL has been observed in E2 subjects (Miettinen et al. Reference Miettinen, Gylling, Vanhanen and Ollus1992; Gylling et al. Reference Gylling, Kontula and Miettinen1995). In addition, ε2 allele carriers have been associated with lower intestinal cholesterol absorption and higher bile acid synthesis than E3 or E4 individuals (Kesaniemi et al. Reference Kesaniemi, Ehnholm and Miettinen1987; Miettinen et al. Reference Miettinen, Gylling, Vanhanen and Ollus1992; Gylling et al. Reference Gylling, Kontula and Miettinen1995). However, these results have been challenged by Von Bergman et al. (Reference Von Bergmann, Lutjohann, Lindenthal and Steinmetz2003), who have reported no differences in intestinal cholesterol absorption and synthesis in E2/E2 v. E4/E4 individuals. Also, there is currently no plausible mechanism linking apoE genotype and the efficiency of cholesterol absorption.

What about the higher LDLC levels in ε4 allele carriers? In most studies the differences relative to E3/E3 subjects are not significant, but there is a consistent trend towards higher total cholesterol and LDLC levels in E4 carriers. Although there are no differences in LDL-receptor binding between E4 and E3 individuals, as mentioned previously the amino acid change at position 112 influences the lipoprotein ‘preference’ of the protein, leading to a higher concentration associated with TAG-rich lipoproteins (chylomicrons and VLDL) as compared with E3 homozygotes (Gregg et al. Reference Gregg, Zech, Schaefer, Stark, Wilson and Brewer1986; Weisgraber, Reference Weisgraber1990). More apoE per TAG-rich lipoprotein particle would be anticipated to result in increased competition with LDL for LDL receptor-mediated clearance, which may lead to increased circulating LDLC levels (Jackson et al. Reference Jackson, Maitin, Leake, Yaqoob and Williams2006). In a number of biokinetic studies (Gregg et al. Reference Gregg, Zech, Schaefer, Stark, Wilson and Brewer1986; Demant et al. Reference Demant, Bedford, Packard and Shepherd1991; Welty et al. Reference Welty, Lichtenstein, Barrett, Jenner, Dolnikowski and Schaefer2000) a lower fractional catabolic rate of LDL-apoB100 has been reported in ε4 allele carriers. In addition, an increased conversion of VLDL to LDL-apoB100 was observed in E4 individuals. This increased synthetic rate together with the reported increased intestinal cholesterol absorption efficiency (Kesaniemi et al. Reference Kesaniemi, Ehnholm and Miettinen1987) could contribute to the trends towards higher LDLC levels in E4 carriers.

Regardless of the mechanism for the LDLC-modulating effects, it is evident that the average 8% higher LDLC levels alone cannot explain the disease differential in E4 carriers (Law et al. Reference Law, Wald and Thompson1994). Furthermore, no consistent difference in CVD risk has been observed between E2 carriers and E3/E3 individuals despite the 10–15% lower LDLC levels, which based on predictive equations would be associated with a 20–30% lower CVD risk (Law et al. Reference Law, Wald and Thompson1994). Thus, it is likely that other mechanisms in part mediate the effect of apoE genotype on CVD pathology.

ApoE genotype and other lipid risk factors for CVD

Inconsistent associations between apoE genotype and fasting TAG levels have been reported in the literature (Brown & Roberts, Reference Brown and Roberts1991; Howard et al. Reference Howard, Gidding and Liu1998; Bercedo-Sanz et al. Reference Bercedo-Sanz, Gonzalez-Lamuno, Malaga and Garcia-Fuentes1999; Inamdar et al. Reference Inamdar, Kelkar, Devasagayam and Bapat2000; Szalai et al. Reference Szalai, Czinner and Csaszar2000; Tan et al. Reference Tan, Tai, Tan, Chia, Lee, Chew and Ordovas2003), and a meta-analysis (Dallongeville et al. Reference Dallongeville, Lussier-Cacan and Davignon1992) has concluded that E2/E2, E2/E4 E2/E3 and E3/E4 subgroups have higher fasting TAG levels than E3/E3 individuals. Higher fasting TAG levels are thought to be attributable to the limited receptor affinity of the apoE2 protein present on VLDL remnants resulting in impaired hepatic clearance of TAG-rich lipoproteins. The mechanisms that could potentially contribute to the moderate hypertriacylglycerolaemia evident in E4 carriers are currently unclear.

Plasma TAG levels in the postprandial state (postprandial lipaemia) are recognised to be a stronger determinant of CVD risk relative to fasting TAG levels (Zilversmit, Reference Zilversmit1979; Patsch et al. Reference Patsch, Esterbauer, Foger and Patsch2000). It has been shown (Weintraub et al. Reference Weintraub, Eisenberg and Breslow1987; Dallongeville et al. Reference Dallongeville, Tiret, Visvikis, O'Reilly, Saava, Tsitouris, Rosseneu, DeBacker, Humphries and Beisiegel1999) that E2 carriers have a relatively delayed exaggerated chylomicron remnant clearance and exaggerated lipaemia and a homozygous E2/E2 genotype is one of the recognised causes of a type III hyperbetalipoproteinaemia phenotype. However, the majority of studies (Brenninkmeijer et al. Reference Brenninkmeijer, Stuyt, Demacker, Stalenhoef and van't Laar1987; Weintraub et al. Reference Weintraub, Eisenberg and Breslow1987; Brown & Roberts, Reference Brown and Roberts1991; Boerwinkle et al. Reference Boerwinkle, Brown, Sharrett, Heiss and Patsch1994; Orth et al. Reference Orth, Wahl, Hanisch, Friedrich, Wieland and Luley1996) show that only E2 homozygotes have impaired chylomicron remnant clearance, with one ε3 allele largely compensating for the impaired receptor binding. As for the implication of the ε4 allele in postprandial metabolism, the data published are inconsistent, with only moderate trends towards impaired metabolism observed.

Although apoE is a constituent of HDL, there are much less data available on the effect of apoE genotype on HDL-cholesterol (HDLC) than there are on LDLC levels. In general, there is a trend towards a reduction in circulating HDLC levels from an E2 genotype to an E4 genotype; some studies (Dallongeville et al. Reference Dallongeville, Lussier-Cacan and Davignon1992; Howard et al. Reference Howard, Gidding and Liu1998; Dallongeville et al. Reference Dallongeville, Tiret, Visvikis, O'Reilly, Saava, Tsitouris, Rosseneu, DeBacker, Humphries and Beisiegel1999; Minihane et al. Reference Minihane, Khan, Leigh-Firbank, Talmud, Wright, Murphy, Griffin and Williams2000; Tan et al. Reference Tan, Tai, Tan, Chia, Lee, Chew and Ordovas2003) have reported effects of genotype on HDLC and apoA1 levels, while other studies (Bercedo-Sanz et al. Reference Bercedo-Sanz, Gonzalez-Lamuno, Malaga and Garcia-Fuentes1999; Szalai et al. Reference Szalai, Csaszar, Czinner, Palicz, Halmos and Romics1999; Inamdar et al. Reference Inamdar, Kelkar, Devasagayam and Bapat2000; Sheehan et al. Reference Sheehan, Bennett and Cashman2000) have not demonstrated an association. As fasting TAG levels are known to be an important determinant of HDL metabolism and HDLC levels, it may be predicted that lower HDLC levels may be evident in E2 and E4 carriers. This lack of TAG–HDLC response to genotype suggests that a TAG-independent mechanism may also play a role in modulating the effect of genotype on HDLC.

ApoE genotype and responsiveness to dietary fat manipulation

The influence of environmental factors on genotype–disease associations is being increasingly recognised. A limited number of studies have indicated that alcohol intake influences apoE–CVD associations (Corella et al. Reference Corella, Guillen, Saiz, Portoles, Sabater, Cortina, Folch, Gonzalez and Ordovas2001a,Reference Corella, Tucker, Lahoz, Coltell, Cupples, Wilson, Schaefer and Ordovasb), but the greatest evidence exists for an impact of smoking status and dietary total fat content and fatty acid composition on the LDLC modulatory effects of the apoE genotype. Responsiveness to dietary fat manipulation is recognised to be highly variable, with genetic variability known to be partly responsible. The systematic review by Masson et al. (Reference Masson, McNeill and Avenell2003) includes studies that have examined the impact of genotype on the responsiveness of fasting lipids to dietary cholesterol (fifteen individual studies) and total fat or fatty acid composition (thirty-six individual studies; mainly manipulation of SFA, MUFA and PUFA ratios). Three of the cholesterol-manipulation studies have reported a greater circulating cholesterol response in E4 carriers. Eleven of the studies that manipulated dietary fat have demonstrated a genotype×treatment interaction, with the E4 subgroup being generally the most responsive (Masson et al. Reference Masson, McNeill and Avenell2003). For example, in the Schaefer et al. (Reference Schaefer, Lamon-Fava, Ausman, Ordovas, Clevidence, Judd, Goldin, Woods, Gorbach and Lichtenstein1997) study (n 148) a National Cholesterol Education Program Step 2 diet was found to result in an overall mean reduction in LDLC levels of 19% and 16% in men and women respectively, with corresponding response ranges of +3% to –55% and +13% to –39%. In the male participants, but not in the female participants, an E4 genotype was shown to be associated with greater LDLC reductions. In the systematic review (Masson et al. Reference Masson, McNeill and Avenell2003) the lack of significance reported in many of the studies is likely to be attributable to a lack of power to detect an inter-genotype difference in response, rather than a lack of a ‘real’ biological effect of apoE genotype. Many of the studies included cohorts of less than fifty participants and retrospective apoE genotype profiling, which often resulted in small group sizes in the rare allele groups. Of the eleven studies that reported significant impacts of apoE genotype, six included more than fifty participants, with an additional study that included forty-five participants (n 15 for E3/E3, E3/E4 and E4/E4 groups) prospectively recruiting on the basis of apoE genotype (Sarkkinen et al. Reference Sarkkinen, Korhonen, Erkkila, Ebeling and Uusitupa1998).

It is likely that background dietary fat composition is partly responsible for the variation in associations between apoE genotype and CVD risk and blood lipid profile reported in the literature. Furthermore, in E4 individuals with a high-fat high-cholesterol high-SFA diet dietary fat manipulation may offer a viable means of counteracting the increased CVD risk. However, before this approach can be advocated with any certainty additional adequately-powered studies are needed in order to fully elucidate the impact of apoE genotype on the heterogeneity in response to dietary total fat and SFA, MUFA and PUFA content.

Recent evidence (Minihane et al. Reference Minihane, Khan, Leigh-Firbank, Talmud, Wright, Murphy, Griffin and Williams2000) also suggests that apoE genotype may in part predict the LDLC response to fish oil fatty acid intervention. The variability of LDLC-raising effect of EPA and DHA has been frequently reported (Harris, Reference Harris1997). In a study of individuals with an atherogenic lipoprotein phenotype (Minihane et al. Reference Minihane, Khan, Leigh-Firbank, Talmud, Wright, Murphy, Griffin and Williams2000) retrospective genotyping suggests that the LDLC-raising effects observed following supplementation with 3 g EPA+DHA/d are associated with an apoE4 genotype. Additional studies are currently underway to investigate EPA/DHA–LDLC associations.

Lipoprotein-independent effects of the apoE protein and apoE genotype: impact on macrophage, endothelial cell, smooth muscle cell and platelet function

As mentioned earlier, although an E4 genotype is associated with moderately-higher LDLC and TAG levels and a trend towards lower HDLC levels, these effects alone are unlikely to be responsible for the higher CVD burden, even in individuals with a high total fat and saturated fat intake. It is therefore speculated that lipid-independent mechanisms may contribute substantially to disease risk.

Monocyte-derived macrophages can produce up to 20% of the total apoE (Basu et al. Reference Basu, Brown, Ho, Havel and Goldstein1981, Reference Basu, Ho, Brown, Bilheimer, Anderson and Goldstein1982; Newman et al. Reference Newman, Dawson, Rudel and Williams1985; Wang-Iverson et al. Reference Wang-Iverson, Gibson and Brown1985). The anti-atherogenic roles of macrophage apoE have been demonstrated in apoE-null rodents (Bellosta et al. Reference Bellosta, Mahley, Sanan, Murata, Newland, Taylor and Pitas1995; Thorngate et al. Reference Thorngate, Rudel, Walzem and Williams2000). In these animals low-level tissue-specific expression of human apoE in macrophages inhibits atherogenesis without substantially influencing the plasma lipid profile.

The role of locally-secreted apoE in the artery wall is currently only partly understood, but it has been proposed to exert several biological functions (Fig. 2). Acting as a paracrine agent, macrophage-derived apoE is known to influence smooth muscle cell (Swertfeger & Hui, Reference Swertfeger and Hui2001), endothelial cell (Stannard et al. Reference Stannard, Riddell, Sacre, Tagalakis, Langer, von Eckardstein, Cullen, Athanasopoulos, Dickson and Owen2001), lymphocyte (Mistry et al. Reference Mistry, Clay, Kelly, Steiner and Harmony1995) and platelet (Riddell et al. Reference Riddell, Graham and Owen1997) function. Within the macrophage itself apoE is involved in reverse cholesterol efflux from macrophages (Shimano et al. Reference Shimano, Ohsuga, Shimada, Namba, Gotoda, Harada, Katsuki, Yazaki and Yamada1995) and is known to modulate the cell inflammatory response through an impact on NO and proinflammatory cytokine production (Colton et al. Reference Colton, Czapiga, Snell-Callanan, Chernyshev and Vitek2001, Reference Colton, Brown, Cook, Needham, Xu, Czapiga, Saunders, Schmechel, Rasheed and Vitek2002). Although data is currently lacking, accumulating evidence suggests an impact of apoE genotype on these metabolic processes, which may be attributable partly to differences in the antioxidant capacity of the apoE isoforms.

Fig. 2. Local effects of apoE on the artery wall. M, monocyte; MΦ, macrophage; EC, endothelial cell; P, platelet; T, T lymphocyte; SMC, smooth muscle cells; VCAM-1, vascular cell adhesion molecule-1.

ApoE and platelet aggregation

Desai et al. (Reference Desai, Bruckdorfer, Hutton and Owen1989) have observed that the binding of apoE as a component of large HDL2 particles to saturable sites on platelets is associated with an inhibition of platelet aggregation. More recent studies (Riddell et al. Reference Riddell, Graham and Owen1997, Reference Riddell, Vinogradov, Stannard, Chadwick and Owen1999, Reference Riddell, Sun, Stannard, Soutar and Owen2001) have suggested that apoE may inhibit platelet reactivity by interacting with apoE receptor 2, which would result in an increase in cellular NO levels as a result of simulation of the NO synthase signalling cascade. The impact of apoE genotype on the anti-aggregatory effect of apoE has not been investigated.

ApoE and adhesion molecule expression

In endothelial cells the interaction of apoE with apoE receptor 2 has been proposed to activate NO synthase through an effect on 1-phosphatidylinositol 3-kinase signalling, with a resultant NO-induced inhibition of vascular cell adhesion molecule-1 induction (Stannard et al. Reference Stannard, Riddell, Sacre, Tagalakis, Langer, von Eckardstein, Cullen, Athanasopoulos, Dickson and Owen2001). In a cell-culture model (EAhy926) Sacre et al. (Reference Sacre, Stannard and Owen2003) have observed an isoform-specific induction of endothelial NO in the order E3>E2>E4. The impact of apoE genotype on adhesion molecule expression in vivo is unknown, although a recently-completed study (AM Minihane et al. unpublished results) indicates an effect of apoE genotype on circulating vascular cell adhesion molecule levels in human volunteers, with the relative levels (E4>E3>E2) consistent with the NO induction observed by Sacre et al. (Reference Sacre, Stannard and Owen2003).

ApoE and smooth muscle cell migration and proliferation

Smooth muscle cell migration into the intima and subsequent proliferation are considered to play an important role in atherosclerosis. ApoE has been shown to inhibit platelet-derived growth factor-directed smooth muscle cell migration by binding to LDL receptor-related protein, which activates the cAMP, protein kinase cascade (Hui & Basford, Reference Hui and Basford2005). In addition, apoE inhibits cell proliferation through binding to cell surface proteoglycans, by a mechanism in which inducible NO synthase is increased (Hui & Basford, Reference Hui and Basford2005). It has been demonstrated that the isoforms do not differ in terms of cell migration inhibition, since binding of lipid-free apoE to LDL receptor-related protein does not show isoform preferences (Zeleny et al. Reference Zeleny, Swertfeger, Weisgraber and Hui2002). On the contrary, apoE2 and apoE3 are more efficient in inhibiting smooth muscle cell proliferation than apoE4 (Zeleny et al. Reference Zeleny, Swertfeger, Weisgraber and Hui2002), which is consistent with the different binding capacity of apoE to heparin sulphate proteoglycans (Cullen et al. Reference Cullen, Cignarella, Brennhausen, Mohr, Assmann and von Eckardstein1998; Hara et al. Reference Hara, Matsushima, Satoh, Iso-o, Noto, Togo, Kimura, Hashimoto and Tsukamoto2003).

ApoE and cellular cholesterol efflux and reverse cholesterol transport

The involvement of apoE in mediating cholesterol efflux from macrophages was first identified by Basu et al. (Reference Basu, Ho, Brown, Bilheimer, Anderson and Goldstein1982) and is now supported by several lines of evidence (Shimano et al. Reference Shimano, Ohsuga, Shimada, Namba, Gotoda, Harada, Katsuki, Yazaki and Yamada1995).

ApoE seems to promote cholesterol efflux when endogenously expressed and to a lesser extent when exogenously added (Lin et al. Reference Lin, Duan and Mazzone1999), and it has been hypothesised that the macrophage and non-macrophage apoE act via divergent mechanisms (Lin et al. Reference Lin, Duan and Mazzone1999) that work in parallel (Dove et al. Reference Dove, Linton and Fazio2005). The enhancing effect can be observed in the absence of acceptors (Zhang et al. Reference Zhang, Gaynor and Kruth1996) and in the presence of cholesterol acceptors such as HDL or phospholipid vesicles (Mazzone & Reardon, Reference Mazzone and Reardon1994). There is a very complex literature relating to the mechanisms by which apoE influences cholesterol efflux in macrophages, and several mechanisms have been proposed (Table 5).

Table 5. Proposed roles for apoE in reverse cholesterol transport

The metabolism of cholesterol in macrophages has been found to differ among the three isoforms. In the absence of extracellular acceptors cholesterol-loaded monocyte-derived macrophages isolated from E4/E4 carriers are less effective in cholesterol efflux than E3/E3 cells, which are less effective than E2/E2 cells (Cullen et al. Reference Cullen, Cignarella, Brennhausen, Mohr, Assmann and von Eckardstein1998). In mouse macrophages (RAW 264·7) the efficiency of cholesterol efflux is in the order E2>E3>E4, which is attributed to isoform variations in binding capacities to heparin sulphate proteoglycans. A higher binding activity of apoE4 is considered to result in higher uptake or degradation of apoE, which results in lower cholesterol efflux activity (Hara et al. Reference Hara, Matsushima, Satoh, Iso-o, Noto, Togo, Kimura, Hashimoto and Tsukamoto2003). This lower efficiency of cholesterol efflux in E4 individuals could make an important contribution to the higher CVD burden observed.

ApoE, NO production and inflammatory status

NO is regarded as a potent macrophage pro-inflammatory mediator. The addition of apoE has been shown to increase monocyte-derived macrophage NO production (Colton et al. Reference Colton, Czapiga, Snell-Callanan, Chernyshev and Vitek2001) by increasing the uptake of arginine (the substrate for NO production) as a result of the up-regulation of the cationic acid transporter family (Colton et al. Reference Colton, Czapiga, Snell-Callanan, Chernyshev and Vitek2001). ApoE isoform-mediated differences in monocyte-derived macrophage NO production have been observed in several models, with higher levels of NO produced by apoE4 macrophages compared with apoE3 macrophages (Colton et al. Reference Colton, Needham, Brown, Cook, Rasheed, Burke, Strittmatter, Schmechel and Vitek2004).

In addition to NO, macrophages produce and secrete an array of pro-inflammatory cytokines, including a number of chemokines, which impact on atherogenesis in both an autocrine and paracrine manner. Data on the impact of apoE genotype on the macrophage inflammatory response are very limited. In a recent studies by Ophir et al. (Reference Ophir, Meilin, Efrati, Chapman, Karussis, Roses and Michaelson2003, Reference Ophir, Amariglio, Jacob-Hirsch, Elkon, Rechavi and Michaelson2005) and Lynch et al. (Reference Lynch, Tang, Wang, Vitek, Bennett, Sullivan, Warner and Laskowitz2003) higher production of pro-inflammatory cytokines in the brain and serum was observed in E4 v. E3 transgenic mice following injection with lipopolysaccharide (inflammatory stimulus). The study of Lynch et al. (Reference Lynch, Tang, Wang, Vitek, Bennett, Sullivan, Warner and Laskowitz2003) highlights that the impact of genotype is largely attributable to a differential impact of E3 v. E4 on NF-κB signalling, which may be attributable to apoE genotype-mediated differences in oxidative status.

ApoE genotype and oxidative status

There are several lines of evidence demonstrating that apoE has antioxidant capacity (Hayek et al. Reference Hayek, Oiknine, Brook and Aviram1994; Pratico et al. Reference Pratico, Lee, Trojanowski, Rokach and Fitzgerald1998; Aviram et al. Reference Aviram, Dornfeld, Rosenblat, Volkova, Kaplan, Coleman, Hayek, Presser and Fuhrman2000; Kitagawa et al. Reference Kitagawa, Matsumoto, Kuwabara, Takasawa, Tanaka, Sasaki, Matsushita, Ohtsuki, Yanagihara and Hori2002). Miyata & Smith (Reference Miyata and Smith1996), whilst investigating the impact of apoE genotype on Alzheimer's disease pathology, were the first to propose allele-specific differences in the antioxidant capacities of apoE isoforms in the order E2>E3>E4, with E2 emerging in in vitro systems as having a 2-fold higher antioxidant capacity relative to E4. Subsequent in vitro studies and brain autopsy investigations of patients with Alzheimer's disease (Jolivalt et al. Reference Jolivalt, Leininger-Muller, Bertrand, Herber, Christen and Siest2000; Tamaoka et al. Reference Tamaoka, Miyatake, Matsuno, Ishii, Nagase and Sahara2000) have confirmed these earlier findings.

Indirect but strong evidence for a role of apoE-mediated differences in oxidative stress being important in CVD pathology is provided by two recent prospective cardiovascular surveillance studies, i.e. the Northwick Park Heart Study (Humphries et al. Reference Humphries, Talmud, Hawe, Bolla, Day and Miller2001) and the Framingham Offspring Study (Talmud et al. Reference Talmud, Stephens, Hawe, Demissie, Cupples, Hurel, Humphries and Ordovas2005). Both studies conclude that after correction for classical risk factors (including lipids) an increased risk of CVD in E4 carriers is only evident in those who smoke, which strongly indicates that an impact of genotype on oxidative status is important. The results of the Northwick Park Heart Study are presented in Table 6, with an adjusted (including for blood lipids) hazard ratio of 2·79 in E4 carriers who were smokers compared with a combined genotype non-smoking group. Although no data is currently available, based on the smoking–genotype interaction observed it may also be speculated that the impact of an E4 genotype may be more evident in individuals with a low dietary antioxidant intake.

Table 6. CHD adjusted hazard ratios (HR) according to apoE genotype for men participating in the Northwick Park Heart StudyFootnote * (adapted from Humphries et al. Reference Humphries, Talmud, Hawe, Bolla, Day and Miller2001)

E2 carriers, E2/E2, E2/E3; E4 carriers, E3/E4, E4/E2.

* Results are compared with the never-smokers, all genotypes combined.

Results adjusted for clinic, age, BMI, systolic blood pressure, plasma lipids (cholesterol and TAG) and fibrinogen.

Recent evidence (Dietrich et al. Reference Dietrich, Hua, Block, Olano, Packer, Morrow, Hudes, Abdukeyum, Rimbach and Minihane2005; Jofre-Monseny et al. Reference Jofre-Monseny, de Pascual-Teresa, Plonka, Huebbe, Boesch-Saasatmandi, Minihane and Rimbach2007) supports a role of apoE genotype in mediating oxidative status. In a mixed smoking and non-smoking group 29% higher levels of lipid peroxidation (as measured by circulating F2-isoprostane levels) were observed in individuals with a total plasma cholesterol >5·6 mmol/l (Dietrich et al. Reference Dietrich, Hua, Block, Olano, Packer, Morrow, Hudes, Abdukeyum, Rimbach and Minihane2005). Furthermore, in a murine macrophage (RAW 264.7) cell line stably transfected with the human apoE3 and apoE4 gene it was observed that an apoE4 genotype is associated with increased membrane oxidation and NO and superoxide anion radical production (Jofre-Monseny et al. Reference Jofre-Monseny, de Pascual-Teresa, Plonka, Huebbe, Boesch-Saasatmandi, Minihane and Rimbach2007).

The exact molecular mechanism by which apoE could exert its antioxidant effects and why it is isoform-dependent is not well understood. A number of possible mechanisms have been suggested, including an effect of genotype on protein folding impacting on the metal-binding domain of the protein located in the amino terminal (Miyata & Smith, Reference Miyata and Smith1996; Pham et al. Reference Pham, Kodvawala and Hui2005). Whatever the mechanism, it seems likely that genotype differences in oxidative status, in particular within the microenvironment of the arterial intima, are partly responsible for the higher CVD risk in E4 carriers, and that therapies targeted at reducing oxidative status and its metabolic consequences could help negate the deleterious effects of an apoE genotype.

Conclusion

Although extensively investigated, the role of the apoE protein and the impact of apoE genotype on cardiovascular health and pathology are only partly understood. It is now evident that part of the CVD burden associated with an E4 genotype is independent of an effect on lipoprotein metabolism, with an impact of genotype on oxidative status and macrophage function being increasingly recognised. Furthermore, observational and intervention trials based on retrospective genotyping of the study participants have highlighted the impact of environmental factors such as smoking status and dietary fat composition on genotype–phenotype associations. Further studies using a large-scale retrospective-genotyping approach or a smaller more-focused approach with individuals prospectively recruited on the basis of genotype are needed to establish the potential of different dietary manipulations to counteract the increased CVD risk in E4 carriers (25% of the UK population). However, it is recognised that because of the complexity and cost such an approach cannot be used to investigate all potential genotype–environment–phenotype associations. Human transgenic cells and animal models can provide a useful tool to initially screen potential dietary components of interest.

References

Acharya, P, Segall, ML, Zaiou, M, Morrow, J, Weisgraber, KH, Phillips, MC, Lund-Katz, S & Snow, J (2002) Comparison of the stabilities and unfolding pathways of human apolipoprotein E isoforms by differential scanning calorimetry and circular dichroism. Biochimica et Biophysica Acta 1584, 919.CrossRefGoogle ScholarPubMed
Aguilar, CA, Talavera, G, Ordovas, JM, Barriguete, JA, Guillen, LE, Leco, ME, Pedro-Botet, J, Gonzalez-Barranco, J, Gomez-Perez, FJ & Rull, JA (1999) The apolipoprotein E4 allele is not associated with an abnormal lipid profile in a Native American population following its traditional lifestyle. Atherosclerosis 142, 409414.CrossRefGoogle ScholarPubMed
Almeida, S, Fiegenbaum, M, de Andrade, FM, Osorio-Wender, MC & Hutz, MH (2006) ESR1 and APOE gene polymorphisms, serum lipids, and hormonal replacement therapy. Maturitas 54, 119126.CrossRefGoogle ScholarPubMed
Aviram, M, Dornfeld, L, Rosenblat, M, Volkova, N, Kaplan, M, Coleman, R, Hayek, T, Presser, D & Fuhrman, B (2000) Pomegranate juice consumption reduces oxidative stress, atherogenic modifications to LDL, and platelet aggregation: studies in humans and in atherosclerotic apolipoprotein E-deficient mice. American Journal of Clinical Nutrition 71, 10621076.CrossRefGoogle ScholarPubMed
Basu, SK, Brown, MS, Ho, YK, Havel, RJ & Goldstein, JL (1981) Mouse macrophages synthesize and secrete a protein resembling apolipoprotein E. Proceedings of the National Academy of Sciences USA 78, 75457549.CrossRefGoogle ScholarPubMed
Basu, SK, Ho, YK, Brown, MS, Bilheimer, DW, Anderson, RG & Goldstein, JL (1982) Biochemical and genetic studies of the apoprotein E secreted by mouse macrophages and human monocytes. Journal of Biological Chemistry 257, 97889795.CrossRefGoogle ScholarPubMed
Beilby, JP, Hunt, CC, Palmer, LJ, Chapman, CM, Burley, JP, McQuillan, BM, Thompson, PL & Hung, J (2003) Apolipoprotein E gene polymorphisms are associated with carotid plaque formation but not with intima-media wall thickening: results from the Perth Carotid Ultrasound Disease Assessment Study (CUDAS). Stroke 34, 869874.CrossRefGoogle Scholar
Bellosta, S, Mahley, RW, Sanan, DA, Murata, J, Newland, DL, Taylor, JM & Pitas, RE (1995) Macrophage-specific expression of human apolipoprotein E reduces atherosclerosis in hypercholesterolemic apolipoprotein E-null mice. Journal of Clinical Investigation 96, 21702179.CrossRefGoogle ScholarPubMed
Bercedo-Sanz, A, Gonzalez-Lamuno, D, Malaga, S & Garcia-Fuentes, M (1999) Impact of ApoE4 allele on total cholesterol levels of children in northern Spain. Clinical Genetics 55, 6970.Google ScholarPubMed
Bhatnagar, D & Durrington, P (1993) Does measurement of apolipoproteins add to the clinical diagnosis and management of dyslipidaemias. Current Opinion of Lipidology 4, 299304.CrossRefGoogle Scholar
Blue, ML, Williams, DL, Zucker, S, Khan, SA & Blum, CB (1983) Apolipoprotein E synthesis in human kidney, adrenal gland, and liver. Proceedings of the National Academy of Sciences USA 80, 283287.CrossRefGoogle Scholar
Boerwinkle, E, Brown, S, Sharrett, AR, Heiss, G & Patsch, W (1994) Apolipoprotein E polymorphism influences postprandial retinyl palmitate but not triglyceride concentrations. American Journal of Human Genetics 54, 341360.Google Scholar
Bradley, WA & Gianturco, SH (1986) ApoE is necessary and sufficient for the binding of large triglyceride-rich lipoproteins to the LDL receptor; apoB is unnecessary. Journal of Lipid Research 27, 4048.CrossRefGoogle Scholar
Brenninkmeijer, BJ, Stuyt, PM, Demacker, PN, Stalenhoef, AF & van't Laar, A (1987) Catabolism of chylomicron remnants in normolipidemic subjects in relation to the apoprotein E phenotype. Journal of Lipid Research 28, 361370.CrossRefGoogle Scholar
Brown, AJ & Roberts, DC (1991) The effect of fasting triacylglyceride concentration and apolipoprotein E polymorphism on postprandial lipemia. Arteriosclerosis, Thrombosis, and Vascular Biology 11, 17371744.CrossRefGoogle ScholarPubMed
Chroni, A, Nieland, TJ, Kypreos, KE, Krieger, M & Zannis, VI (2005) SR-BI mediates cholesterol efflux via its interactions with lipid-bound ApoE. Structural mutations in SR-BI diminish cholesterol efflux. Biochemistry 44, 1313213143.CrossRefGoogle ScholarPubMed
Colton, CA, Brown, CM, Cook, D, Needham, LK, Xu, Q, Czapiga, M, Saunders, AM, Schmechel, DE, Rasheed, K & Vitek, MP (2002) APOE and the regulation of microglial nitric oxide production: a link between genetic risk and oxidative stress. Neurobiology of Aging 23, 777785.CrossRefGoogle ScholarPubMed
Colton, CA, Czapiga, M, Snell-Callanan, J, Chernyshev, ON & Vitek, MP (2001) Apolipoprotein E acts to increase nitric oxide production in macrophages by stimulating arginine transport. Biochimica et Biophysica Acta 1535, 134144.CrossRefGoogle ScholarPubMed
Colton, CA, Needham, LK, Brown, C, Cook, D, Rasheed, K, Burke, JR, Strittmatter, WJ, Schmechel, DE & Vitek, MP (2004) APOE genotype-specific differences in human and mouse macrophage nitric oxide production. Journal of Neuroimmunology 147, 6267.CrossRefGoogle ScholarPubMed
Corbo, RM & Scacchi, R (1999) Apolipoprotein E (APOE) allele distribution in the world. Is APOE*4 a ‘thrifty’ allele? Annals of Human Genetics 63, 301310.CrossRefGoogle Scholar
Corella, D, Guillen, M, Saiz, C, Portoles, O, Sabater, A, Cortina, S, Folch, J, Gonzalez, JI & Ordovas, JM (2001 a) Environmental factors modulate the effect of the APOE genetic polymorphism on plasma lipid concentrations: ecogenetic studies in a Mediterranean Spanish population. Metabolism 50, 936944.CrossRefGoogle Scholar
Corella, D, Tucker, K, Lahoz, C, Coltell, O, Cupples, LA, Wilson, PW, Schaefer, EJ & Ordovas, JM (2001 b) Alcohol drinking determines the effect of the APOE locus on LDL-cholesterol concentrations in men: the Framingham Offspring Study. American Journal of Clinical Nutrition 73, 736745.CrossRefGoogle ScholarPubMed
Cullen, P, Cignarella, A, Brennhausen, B, Mohr, S, Assmann, G & von Eckardstein, A (1998) Phenotype-dependent differences in apolipoprotein E metabolism and in cholesterol homeostasis in human monocyte-derived macrophages. Journal of Clinical Investigation 101, 16701677.CrossRefGoogle ScholarPubMed
Dallongeville, J, Lussier-Cacan, S & Davignon, J (1992) Modulation of plasma triglyceride levels by apoE phenotype: a meta-analysis. Journal of Lipid Research 33, 447454.CrossRefGoogle ScholarPubMed
Dallongeville, J, Tiret, L, Visvikis, S, O'Reilly, DS, Saava, M, Tsitouris, G, Rosseneu, M, DeBacker, G, Humphries, SE & Beisiegel, U (1999) Effect of apo E phenotype on plasma postprandial triglyceride levels in young male adults with and without a familial history of myocardial infarction: the EARS II study. European Atherosclerosis Research Study. Atherosclerosis 145, 381388.CrossRefGoogle ScholarPubMed
Davignon, J, Gregg, RE & Sing, CF (1988) Apolipoprotein E polymorphism and atherosclerosis. Arteriosclerosis 8, 121.CrossRefGoogle ScholarPubMed
Dawson, PA, Lukaszewski, LM, Ells, PF, Malbon, CC & Williams, DL (1989) Quantification and regulation of apolipoprotein E expression in rat Kupffer cells. Journal of Lipid Research 30, 403413.CrossRefGoogle ScholarPubMed
Demant, T, Bedford, D, Packard, CJ & Shepherd, J (1991) Influence of apolipoprotein E polymorphism on apolipoprotein B-100 metabolism in normolipemic subjects. Journal of Clinical Investigation 88, 14901501.CrossRefGoogle ScholarPubMed
Desai, K, Bruckdorfer, KR, Hutton, RA & Owen, JS (1989) Binding of apoE-rich high density lipoprotein particles by saturable sites on human blood platelets inhibits agonist-induced platelet aggregation. Journal of Lipid Research 30, 831840.CrossRefGoogle ScholarPubMed
Dietrich, M, Hua, Y, Block, G, Olano, E, Packer, L, Morrow, JD, Hudes, M, Abdukeyum, G, Rimbach, G & Minihane, AM (2005) Associations between apolipoprotein E genotype and circulating F2-isoprostane levels in humans. Lipids 40, 329334.CrossRefGoogle ScholarPubMed
Dong, LM & Weisgraber, KH (1996) Human apolipoprotein E4 domain interaction. Arginine 61 and glutamic acid 255 interact to direct the preference for very low density lipoproteins. Journal of Biological Chemistry 271, 1905319057.CrossRefGoogle ScholarPubMed
Dove, DE, Linton, MF & Fazio, S (2005) ApoE-mediated cholesterol efflux from macrophages: separation of autocrine and paracrine effects. American Journal of Physiology 288, C586C592.CrossRefGoogle ScholarPubMed
Eichner, JE, Dunn, ST, Perveen, G, Thompson, DM, Stewart, KE & Stroehla, BC (2002) Apolipoprotein E polymorphism and cardiovascular disease: a HuGE review. American Journal of Epidemiology 155, 487495.CrossRefGoogle ScholarPubMed
Elosua, R, Ordovas, JM, Cupples, LA, Fox, CS, Polak, JF, Wolf, PA, SrD'Agostino, RA & O'Donnell, CJ (2004) Association of APOE genotype with carotid atherosclerosis in men and women: the Framingham Heart Study. Journal of Lipid Research 45, 18681875.CrossRefGoogle ScholarPubMed
Fazio, S & Yao, Z (1995) The enhanced association of apolipoprotein E with apolipoprotein B-containing lipoproteins in serum-stimulated hepatocytes occurs intracellularly. Arteriosclerosis, Thrombosis, and Vascular Biology 15, 593600.CrossRefGoogle ScholarPubMed
Gregg, RE & Brewer, HB Jr (1988) The role of apolipoprotein E and lipoprotein receptors in modulating the in vivo metabolism of apolipoprotein B-containing lipoproteins in humans. Clinical Chemistry 34, B28B32.Google ScholarPubMed
Gregg, RE, Zech, LA, Schaefer, EJ, Stark, D, Wilson, D & Brewer, HB Jr (1986) Abnormal in vivo metabolism of apolipoprotein E4 in humans. Journal of Clinical Investigation 78, 815821.CrossRefGoogle ScholarPubMed
Gylling, H, Kontula, K & Miettinen, TA (1995) Cholesterol absorption and metabolism and LDL kinetics in healthy men with different apoprotein E phenotypes and apoprotein B Xba I and LDL receptor Pvu II genotypes. Arteriosclerosis, Thrombosis, and Vascular Biology 15, 208213.CrossRefGoogle ScholarPubMed
Hallman, DM, Boerwinkle, E, Saha, N, Sandholzer, C, Menzel, HJ, Csazar, A & Utermann, G (1991) The apolipoprotein E polymorphism: a comparison of allele frequencies and effects in nine populations. American Journal of Human Genetics 49, 338349.Google ScholarPubMed
Hara, M, Matsushima, T, Satoh, H, Iso-o, N, Noto, H, Togo, M, Kimura, S, Hashimoto, Y & Tsukamoto, K (2003) Isoform-dependent cholesterol efflux from macrophages by apolipoprotein E is modulated by cell surface proteoglycans. Arteriosclerosis, Thrombosis, and Vascular Biology 23, 269274.CrossRefGoogle ScholarPubMed
Harris, WS (1997) n-3 fatty acids and serum lipoproteins: human studies. American Journal of Clinical Nutrition 65, 1645S1654S.CrossRefGoogle ScholarPubMed
Hatters, DM, Peters-Libeu, CA & Weisgraber, KH (2006) Apolipoprotein E structure: insights into function. Trends in Biochemical Sciences 31, 445454.CrossRefGoogle ScholarPubMed
Hayek, T, Oiknine, J, Brook, JG & Aviram, M (1994) Increased plasma and lipoprotein lipid peroxidation in apo E-deficient mice. Biochemical and Biophysical Research Communications 201, 15671574.CrossRefGoogle ScholarPubMed
Howard, BV, Gidding, SS & Liu, K (1998) Association of apolipoprotein E phenotype with plasma lipoproteins in African-American and white young adults. The CARDIA Study. Coronary Artery Risk Development in Young Adults. American Journal of Epidemiology 148, 859868.CrossRefGoogle ScholarPubMed
Huang, Y, Ji, ZS, Brecht, WJ, Rall, SC Jr, Taylor, JM & Mahley, RW (1999) Overexpression of apolipoprotein E3 in transgenic rabbits causes combined hyperlipidemia by stimulating hepatic VLDL production and impairing VLDL lipolysis. Arteriosclerosis, Thrombosis, and Vascular Biology 19, 29522959.CrossRefGoogle ScholarPubMed
Hui, DY & Basford, JE (2005) Distinct signaling mechanisms for apoE inhibition of cell migration and proliferation. Neurobiology of Aging 26, 317323.CrossRefGoogle ScholarPubMed
Humphries, SE, Talmud, PJ, Hawe, E, Bolla, M, Day, IN & Miller, GJ (2001) Apolipoprotein E4 and coronary heart disease in middle-aged men who smoke: a prospective study. Lancet 358, 115119.CrossRefGoogle ScholarPubMed
Ilveskoski, E, Perola, M, Lehtimaki, T, Laippala, P, Savolainen, V, Pajarinen, J et al. (1999) Age-dependent association of apolipoprotein E genotype with coronary and aortic atherosclerosis in middle-aged men: an autopsy study. Circulation 100, 608613.CrossRefGoogle ScholarPubMed
Inamdar, PA, Kelkar, SM, Devasagayam, TP & Bapat, MM (2000) Apolipoprotein E polymorphism in non-insulin-dependent diabetics of Mumbai, India and its effect on plasma lipids and lipoproteins. Diabetes Research and Clinical Practice 47, 217223.CrossRefGoogle ScholarPubMed
Innerarity, TL, Friedlander, EJ, Rall, SC Jr, Weisgraber, KH & Mahley, RW (1983) The receptor-binding domain of human apolipoprotein E. Binding of apolipoprotein E fragments. Journal of Biological Chemistry 258, 1234112347.CrossRefGoogle ScholarPubMed
Ishida, BY, Bailey, KR, Duncan, KG, Chalkley, RJ, Burlingame, AL, Kane, JP & Schwartz, DM (2004) Regulated expression of apolipoprotein E by human retinal pigment epithelial cells. Journal of Lipid Research 45, 263271.CrossRefGoogle ScholarPubMed
Jackson, KG, Maitin, V, Leake, DS, Yaqoob, P & Williams, CM (2006) Saturated fat-induced changes in Sf 60–400 particle composition reduces uptake of LDL by HepG2 cells. Journal of Lipid Research 47, 393403.CrossRefGoogle ScholarPubMed
Jarvik, GP, Austin, MA, Fabsitz, RR, Auwerx, J, Reed, T, Christian, JC & Deeb, S (1994) Genetic influences on age-related change in total cholesterol, low density lipoprotein-cholesterol, and triglyceride levels: longitudinal apolipoprotein E genotype effects. Genetic Epidemiology 11, 375384.CrossRefGoogle ScholarPubMed
Jofre-Monseny, L, de Pascual-Teresa, S, Plonka, E, Huebbe, P, Boesch-Saasatmandi, C, Minihane, AM & Rimbach, G (2007) Differential effects of apolipoprotein E3 and E4 on markers of oxidative status in macrophages. British Journal of Nutrition (In the Press).CrossRefGoogle ScholarPubMed
Jolivalt, C, Leininger-Muller, B, Bertrand, P, Herber, R, Christen, Y & Siest, G (2000) Differential oxidation of apolipoprotein E isoforms and interaction with phospholipids. Free Radical Biology and Medicine 28, 129140.CrossRefGoogle ScholarPubMed
Kayden, HJ, Maschio, F & Traber, MG (1985) The secretion of apolipoprotein E by human monocyte-derived macrophages. Archives of Biochemistry and Biophysics 239, 388395.CrossRefGoogle ScholarPubMed
Kesaniemi, YA, Ehnholm, C & Miettinen, TA (1987) Intestinal cholesterol absorption efficiency in man is related to apoprotein E phenotype. Journal of Clinical Investigation 80, 578581.CrossRefGoogle Scholar
Kim, DH, Iijima, H, Goto, K, Sakai, J, Ishii, H, Kim, HJ, Suzuki, H, Kondo, H, Saeki, S & Yamamoto, T (1996) Human apolipoprotein E receptor 2. A novel lipoprotein receptor of the low density lipoprotein receptor family predominantly expressed in brain. Journal of Biological Chemistry 271, 83738380.CrossRefGoogle ScholarPubMed
Kitagawa, K, Matsumoto, M, Kuwabara, K, Takasawa, K, Tanaka, S, Sasaki, T, Matsushita, K, Ohtsuki, T, Yanagihara, T & Hori, M (2002) Protective effect of apolipoprotein E against ischemic neuronal injury is mediated through antioxidant action. Journal of Neuroscience Research 68, 226232.CrossRefGoogle ScholarPubMed
Kuusisto, J, Mykkanen, L, Kervinen, K, Kesaniemi, YA & Laakso, M (1995) Apolipoprotein E4 phenotype is not an important risk factor for coronary heart disease or stroke in elderly subjects. Arteriosclerosis, Thrombosis, and Vascular Biology 15, 12801286.CrossRefGoogle ScholarPubMed
Lahoz, C, Schaefer, EJ, Cupples, LA, Wilson, PW, Levy, D, Osgood, D, Parpos, S, Pedro-Botet, J, Daly, JA & Ordovas, JM (2001) Apolipoprotein E genotype and cardiovascular disease in the Framingham Heart Study. Atherosclerosis 154, 529537.CrossRefGoogle ScholarPubMed
Law, MR, Wald, NJ & Thompson, SG (1994) By how much and how quickly does reduction in serum cholesterol concentration lower risk of ischaemic heart disease? British Medical Journal 308, 367372.CrossRefGoogle ScholarPubMed
Lehtimaki, T, Metso, S, Ylitalo, R, Rontu, R, Nikkila, M, Wuolijoki, E & Ylitalo, P (2005) Microcrystalline chitosan is ineffective to decrease plasma lipids in both apolipoprotein E epsilon 4 carriers and non-carriers: a long-term placebo-controlled trial in hypercholesterolaemic volunteers. Basic & Clinical Pharmacology & Toxicology 97, 98103.CrossRefGoogle ScholarPubMed
Lenzen, HJ, Assmann, G, Buchwalsky, R & Schulte, H (1986) Association of apolipoprotein E polymorphism, low-density lipoprotein cholesterol, and coronary artery disease. Clinical Chemistry 32, 778781.CrossRefGoogle ScholarPubMed
Li, X, Kypreos, K, Zanni, EE & Zannis, V (2003) Domains of apoE required for binding to apoE receptor 2 and to phospholipids: implications for the functions of apoE in the brain. Biochemistry 42, 1040610417.CrossRefGoogle ScholarPubMed
Libeu, CP, Lund-Katz, S, Phillips, MC, Wehrli, S, Hernaiz, MJ, Capila, I, Linhardt, RJ, Raffai, RL, Newhouse, YM, Zhou, F & Weisgraber, KH (2001) New insights into the heparan sulfate proteoglycan-binding activity of apolipoprotein E. Journal of Biological Chemistry 276, 3913839144.CrossRefGoogle ScholarPubMed
Lin, CY, Duan, H & Mazzone, T (1999) Apolipoprotein E-dependent cholesterol efflux from macrophages: kinetic study and divergent mechanisms for endogenous versus exogenous apolipoprotein E. Journal of Lipid Research 40, 16181627.CrossRefGoogle ScholarPubMed
Lynch, JR, Tang, W, Wang, H, Vitek, MP, Bennett, ER, Sullivan, PM, Warner, DS & Laskowitz, DT (2003) APOE genotype and an ApoE-mimetic peptide modify the systemic and central nervous system inflammatory response. Journal of Biological Chemistry 278, 4852948533.CrossRefGoogle Scholar
Mahley, RW (1988) Apolipoprotein E: cholesterol transport protein with expanding role in cell biology. Science 240, 622630.CrossRefGoogle ScholarPubMed
Mahley, RW & Ji, ZS (1999) Remnant lipoprotein metabolism: key pathways involving cell-surface heparan sulfate proteoglycans and apolipoprotein E. Journal of Lipid Research 40, 116.CrossRefGoogle ScholarPubMed
Marques-Vidal, P, Bongard, V, Ruidavets, JB, Fauvel, J, Hanaire-Broutin, H, Perret, B & Ferrieres, J (2003) Obesity and alcohol modulate the effect of apolipoprotein E polymorphism on lipids and insulin. Obesity Research 11, 12001206.CrossRefGoogle Scholar
Masson, LF, McNeill, G & Avenell, A (2003) Genetic variation and the lipid response to dietary intervention: a systematic review. American Journal of Clinical Nutrition 77, 10981111.CrossRefGoogle ScholarPubMed
Mazzone, T & Reardon, C (1994) Expression of heterologous human apolipoprotein E by J774 macrophages enhances cholesterol efflux to HDL3. Journal of Lipid Research 35, 13451353.CrossRefGoogle ScholarPubMed
Miettinen, TA, Gylling, H, Vanhanen, H & Ollus, A (1992) Cholesterol absorption, elimination, and synthesis related to LDL kinetics during varying fat intake in men with different apoprotein E phenotypes. Arteriosclerosis, Thrombosis, and Vascular Biology 12, 10441052.CrossRefGoogle ScholarPubMed
Miltiadous, G, Hatzivassiliou, M, Liberopoulos, E, Bairaktari, E, Tselepis, A, Cariolou, M & Elisaf, M (2005) Gene polymorphisms affecting HDL-cholesterol levels in the normolipidemic population. Nutrition, Metabolism and Cardiovascular Diseases 15, 219224.CrossRefGoogle ScholarPubMed
Minihane, AM, Khan, S, Leigh-Firbank, EC, Talmud, P, Wright, JW, Murphy, MC, Griffin, BA & Williams, CM (2000) ApoE polymorphism and fish oil supplementation in subjects with an atherogenic lipoprotein phenotype. Arteriosclerosis, Thrombosis, and Vascular Biology 20, 19901997.CrossRefGoogle ScholarPubMed
Mistry, MJ, Clay, MA, Kelly, ME, Steiner, MA & Harmony, JA (1995) Apolipoprotein E restricts interleukin-dependent T lymphocyte proliferation at the G1A/G1B boundary. Cellular Immunology 160, 1423.CrossRefGoogle ScholarPubMed
Miyata, M & Smith, JD (1996) Apolipoprotein E allele-specific antioxidant activity and effects on cytotoxicity by oxidative insults and beta-amyloid peptides. Nature Genetics 14, 5561.CrossRefGoogle ScholarPubMed
Morrow, JA, Hatters, DM, Lu, B, Hochtl, P, Oberg, KA, Rupp, B & Weisgraber, KH (2002) Apolipoprotein E4 forms a molten globule. A potential basis for its association with disease. Journal of Biological Chemistry 277, 5038050385.CrossRefGoogle ScholarPubMed
National Center for Biotechnology Information (2006) Single nucleotide polymorphism. www.ncbi.nlm.nih.gov/SNP/ (accessed November 2006)Google Scholar
Newman, TC, Dawson, PA, Rudel, LL & Williams, DL (1985) Quantitation of apolipoprotein E mRNA in the liver and peripheral tissues of nonhuman primates. Journal of Biological Chemistry 260, 24522457.CrossRefGoogle ScholarPubMed
Nolte, RT & Atkinson, D (1992) Conformational analysis of apolipoprotein A-I and E-3 based on primary sequence and circular dichroism. Biophysical Journal 63, 12211239.CrossRefGoogle ScholarPubMed
Ophir, G, Amariglio, N, Jacob-Hirsch, J, Elkon, R, Rechavi, G & Michaelson, DM (2005) Apolipoprotein E4 enhances brain inflammation by modulation of the NF-kappaB signaling cascade. Neurobiology of Disease 20, 709718.CrossRefGoogle ScholarPubMed
Ophir, G, Meilin, S, Efrati, M, Chapman, J, Karussis, D, Roses, A & Michaelson, DM (2003) Human apoE3 but not apoE4 rescues impaired astrocyte activation in apoE null mice. Neurobiology of Disease 12, 5664.CrossRefGoogle Scholar
Orth, M, Wahl, S, Hanisch, M, Friedrich, I, Wieland, H & Luley, C (1996) Clearance of postprandial lipoproteins in normolipemics: role of the apolipoprotein E phenotype. Biochimica et Biophysica Acta 1303, 2230.CrossRefGoogle ScholarPubMed
Pablos-Mendez, A, Mayeux, R, Ngai, C, Shea, S & Berglund, L (1997) Association of apo E polymorphism with plasma lipid levels in a multiethnic elderly population. Arteriosclerosis, Thrombosis, and Vascular Biology 17, 35343541.CrossRefGoogle Scholar
Paik, YK, Chang, DJ, Reardon, CA, Davies, GE, Mahley, RW & Taylor, JM (1985) Nucleotide sequence and structure of the human apolipoprotein E gene. Proceedings of the National Academy of Sciences USA 82, 34453449.CrossRefGoogle ScholarPubMed
Patsch, W, Esterbauer, H, Foger, B & Patsch, JR (2000) Postprandial lipemia and coronary risk. Current Atherosclerosis Reports 2, 232242.CrossRefGoogle ScholarPubMed
Pham, T, Kodvawala, A & Hui, DY (2005) The receptor binding domain of apolipoprotein e is responsible for its antioxidant activity. Biochemistry 44, 75777582.CrossRefGoogle ScholarPubMed
Polacek, D, Beckmann, MW & Schreiber, JR (1992) Rat ovarian apolipoprotein E: localization and gonadotropic control of messenger RNA. Biology of Reproduction 46, 6572.CrossRefGoogle ScholarPubMed
Pratico, D, Lee, VM-Y, Trojanowski, JQ, Rokach, J & Fitzgerald, GA (1998) Increased F2-isoprostanes in Alzheimer's disease: evidence for enhanced lipid peroxidation in vivo. FASEB Journal 12, 17771783.CrossRefGoogle ScholarPubMed
Rall, SC Jr, Weisgraber, KH & Mahley, RW (1982) Human apolipoprotein E. The complete amino acid sequence. Journal of Biological Chemistry 257, 41714178.CrossRefGoogle ScholarPubMed
Ranjith, N, Pegoraro, RJ, Rom, L, Rajput, MC & Naidoo, DP (2004) Lp(a) and apoE polymorphisms in young South African Indians with myocardial infarction. Cardiovascular Journal of South Africa 15, 111117.Google ScholarPubMed
Rastas, S, Mattila, K, Verkkoniemi, A, Niinisto, L, Juva, K, Sulkava, R & Lansimies, E (2004) Association of apolipoprotein E genotypes, blood pressure, blood lipids and ECG abnormalities in a general population aged 85+. BMC Geriatrics 4, 1.CrossRefGoogle Scholar
Remaley, AT, Stonik, JA, Demosky, SJ, Neufeld, EB, Bocharov, AV, Vishnyakova, TG, Eggerman, TL, Patterson, AP, Duverger, NJ, Santamarina-Fojo, S & Brewer, HB Jr (2001) Apolipoprotein specificity for lipid efflux by the human ABCAI transporter. Biochemical and Biophysical Research Communications 280, 818823.CrossRefGoogle ScholarPubMed
Riddell, DR, Graham, A & Owen, JS (1997) Apolipoprotein E inhibits platelet aggregation through the L-arginine:nitric oxide pathway. Implications for vascular disease. Journal of Biological Chemistry 272, 8995.CrossRefGoogle ScholarPubMed
Riddell, DR, Sun, XM, Stannard, AK, Soutar, AK & Owen, JS (2001) Localization of apolipoprotein E receptor 2 to caveolae in the plasma membrane. Journal of Lipid Research 42, 9981002.CrossRefGoogle ScholarPubMed
Riddell, DR, Vinogradov, DV, Stannard, AK, Chadwick, N & Owen, JS (1999) Identification and characterization of LRP8 (apoER2) in human blood platelets. Journal of Lipid Research 40, 19251930.CrossRefGoogle ScholarPubMed
Ruiz, J, Kouiavskaia, D, Migliorini, M, Robinson, S, Saenko, EL, Gorlatova, N, Li, D, Lawrence, D, Hyman, BT, Weisgraber, KH & Strickland, DK (2005) The apoE isoform binding properties of the VLDL receptor reveal marked differences from LRP and the LDL receptor. Journal of Lipid Research 46, 17211731.CrossRefGoogle ScholarPubMed
Sacre, SM, Stannard, AK & Owen, JS (2003) Apolipoprotein E (apoE) isoforms differentially induce nitric oxide production in endothelial cells. FEBS Letters 540, 181187.CrossRefGoogle ScholarPubMed
Saito, H, Dhanasekaran, P, Baldwin, F, Weisgraber, KH, Phillips, MC & Lund-Katz, S (2003) Effects of polymorphism on the lipid interaction of human apolipoprotein E. Journal of Biological Chemistry 278, 4072340729.CrossRefGoogle ScholarPubMed
Saito, M, Eto, M, Nitta, H, Kanda, Y, Shigeto, M, Nakayama, K et al. (2004) Effect of apolipoprotein E4 allele on plasma LDL cholesterol response to diet therapy in type 2 diabetic patients. Diabetes Care 27, 12761280.CrossRefGoogle ScholarPubMed
Sanada, M, Nakagawa, H, Kodama, I, Sakasita, T & Ohama, K (1998) Apolipoprotein E phenotype associations with plasma lipoproteins and bone mass in postmenopausal women. Climacteric 1, 188195.CrossRefGoogle ScholarPubMed
Sarkkinen, E, Korhonen, M, Erkkila, A, Ebeling, T & Uusitupa, M (1998) Effect of apolipoprotein E polymorphism on serum lipid response to the separate modification of dietary fat and dietary cholesterol. American Journal of Clinical Nutrition 68, 12151222.CrossRefGoogle Scholar
Schaefer, EJ, Gregg, RE, Ghiselli, G, Forte, TM, Ordovas, JM, Zech, LA & Brewer, HB Jr (1986) Familial apolipoprotein E deficiency. Journal of Clinical Investigation 78, 12061219.CrossRefGoogle ScholarPubMed
Schaefer, EJ, Lamon-Fava, S, Ausman, LM, Ordovas, JM, Clevidence, BA, Judd, JT, Goldin, BR, Woods, M, Gorbach, S & Lichtenstein, AH (1997) Individual variability in lipoprotein cholesterol response to National Cholesterol Education Program Step 2 diets. American Journal of Clinical Nutrition 65, 823830.CrossRefGoogle ScholarPubMed
Schaefer, EJ, Lamon-Fava, S, Johnson, S, Ordovas, JM, Schaefer, MM, Castelli, WP & Wilson, PW (1994) Effects of gender and menopausal status on the association of apolipoprotein E phenotype with plasma lipoprotein levels. Results from the Framingham Offspring Study. Arteriosclerosis, Thrombosis, and Vascular Biology 14, 11051113.CrossRefGoogle ScholarPubMed
Scuteri, A, Najjar, SS, Muller, D, Andres, R, Morrell, CH, Zonderman, AB & Lakatta, EG (2005) apoE4 allele and the natural history of cardiovascular risk factors. American Journal of Physiology 289, E322E327.Google ScholarPubMed
Sheehan, D, Bennett, T & Cashman, K (2000) Apolipoprotein E gene polymorphisms and serum cholesterol in healthy Irish adults: a proposed genetic marker for coronary artery disease risk. Irish Journal of Medical Sciences 169, 5054.CrossRefGoogle ScholarPubMed
Shimano, H, Ohsuga, J, Shimada, M, Namba, Y, Gotoda, T, Harada, K, Katsuki, M, Yazaki, Y & Yamada, N (1995) Inhibition of diet-induced atheroma formation in transgenic mice expressing apolipoprotein E in the arterial wall. Journal of Clinical Investigation 95, 469476.CrossRefGoogle ScholarPubMed
Shin, MH, Kim, HN, Cui, LH, Kweon, SS, Park, KS, Heo, H, Nam, HS, Jeong, SK, Chung, EK & Choi, JS (2005) The effect of apolipoprotein E polymorphism on lipid levels in Korean adults. Journal of Korean Medical Science 20, 361366.CrossRefGoogle ScholarPubMed
Shore, VG & Shore, B (1973) Heterogeneity of human plasma very low density lipoproteins. Separation of species differing in protein components. Biochemistry 12, 502507.CrossRefGoogle ScholarPubMed
Singh, P, Singh, M & Mastana, S (2006) APOE distribution in world populations with new data from India and the UK. Annals of Human Biology 33, 297308.CrossRefGoogle ScholarPubMed
Song, Y, Stampfer, MJ & Liu, S (2004) Meta-analysis: apolipoprotein E genotypes and risk for coronary heart disease. Annals of Internal Medicine 141, 137147.CrossRefGoogle ScholarPubMed
Srinivasan, SR, Ehnholm, C, Elkasabany, A & Berenson, G (1999) Influence of apolipoprotein E polymorphism on serum lipids and lipoprotein changes from childhood to adulthood: the Bogalusa Heart Study. Atherosclerosis 143, 435443.CrossRefGoogle ScholarPubMed
Stannard, AK, Riddell, DR, Sacre, SM, Tagalakis, AD, Langer, C, von Eckardstein, A, Cullen, P, Athanasopoulos, T, Dickson, G & Owen, JS (2001) Cell-derived apolipoprotein E (ApoE) particles inhibit vascular cell adhesion molecule-1 (VCAM-1) expression in human endothelial cells. Journal of Biological Chemistry 276, 4601146016.CrossRefGoogle ScholarPubMed
Strickland, DK, Gonias, SL & Argraves, WS (2002) Diverse roles for the LDL receptor family. Trends in Endocrinology and Metabolism 13, 6674.CrossRefGoogle ScholarPubMed
Swertfeger, DK & Hui, DY (2001) Apolipoprotein E receptor binding versus heparan sulfate proteoglycan binding in its regulation of smooth muscle cell migration and proliferation. Journal of Biological Chemistry 276, 2504325048.CrossRefGoogle ScholarPubMed
Szalai, C, Csaszar, A, Czinner, A, Palicz, T, Halmos, B & Romics, L (1999) Genetic investigation of patients with hypercholesterolemia type IIa. Clinical Genetics 55, 6768.Google ScholarPubMed
Szalai, C, Czinner, A & Csaszar, A (2000) Influence of apolipoprotein E genotypes on serum lipid parameters in a biracial sample of children. European Journal of Pediatrics 159, 257260.CrossRefGoogle Scholar
Talmud, PJ, Stephens, JW, Hawe, E, Demissie, S, Cupples, LA, Hurel, SJ, Humphries, SE & Ordovas, JM (2005) The significant increase in cardiovascular disease risk in APOEepsilon4 carriers is evident only in men who smoke: potential relationship between reduced antioxidant status and ApoE4. Annals of Human Genetics 69, 613622.CrossRefGoogle ScholarPubMed
Tamaoka, A, Miyatake, F, Matsuno, S, Ishii, K, Nagase, S, Sahara, N et al. (2000) Apolipoprotein E allele-dependent antioxidant activity in brains with Alzheimer's disease. Neurology 54, 23192321.CrossRefGoogle ScholarPubMed
Tan, CE, Tai, ES, Tan, CS, Chia, KS, Lee, J, Chew, SK & Ordovas, JM (2003) APOE polymorphism and lipid profile in three ethnic groups in the Singapore population. Atherosclerosis 170, 253260.CrossRefGoogle ScholarPubMed
Terry, JG, Howard, G, Mercuri, M, Bond, MG & Crouse, JR 3rd (1996) Apolipoprotein E polymorphism is associated with segment-specific extracranial carotid artery intima-media thickening. Stroke 27, 17551759.CrossRefGoogle ScholarPubMed
Thorngate, FE, Rudel, LL, Walzem, RL & Williams, DL (2000) Low levels of extrahepatic nonmacrophage ApoE inhibit atherosclerosis without correcting hypercholesterolemia in ApoE-deficient mice. Arteriosclerosis, Thrombosis, and Vascular Biology 20, 19391945.CrossRefGoogle ScholarPubMed
Utermann, G, Hardewig, A & Zimmer, F (1984) Apolipoprotein E phenotypes in patients with myocardial infarction. Human Genetics 65, 237241.CrossRefGoogle ScholarPubMed
Von Bergmann, K, Lutjohann, D, Lindenthal, B & Steinmetz, A (2003) Efficiency of intestinal cholesterol absorption in humans is not related to apoE phenotype. Journal of Lipid Research 44, 193197.CrossRefGoogle Scholar
Wallis, SC, Rogne, S, Gill, L, Markham, A, Edge, M, Woods, D, Williamson, R & Humphries, S (1983) The isolation of cDNA clones for human apolipoprotein E and the detection of apoE RNA in hepatic and extra-hepatic tissues. The EMBO Journal 2, 23692373.CrossRefGoogle ScholarPubMed
Wang-Iverson, P, Gibson, JC & Brown, WV (1985) Plasma apolipoprotein secretion by human monocyte-derived macrophages. Biochimica et Biophysica Acta 834, 256262.CrossRefGoogle ScholarPubMed
Weintraub, MS, Eisenberg, S & Breslow, JL (1987) Dietary fat clearance in normal subjects is regulated by genetic variation in apolipoprotein E. Journal of Clinical Investigation 80, 15711577.CrossRefGoogle ScholarPubMed
Weisgraber, KH (1990) Apolipoprotein E distribution among human plasma lipoproteins: role of the cysteine-arginine interchange at residue 112. Journal of Lipid Research 31, 15031511.CrossRefGoogle ScholarPubMed
Weisgraber, KH (1994) Apolipoprotein E: structure-function relationships. Advances in Protein Chemistry 45, 249302.CrossRefGoogle ScholarPubMed
Weisgraber, KH, Innerarity, TL & Mahley, RW (1982) Abnormal lipoprotein receptor-binding activity of the human E apoprotein due to cysteine-arginine interchange at a single site. Journal of Biological Chemistry 257, 25182521.CrossRefGoogle Scholar
Weisgraber, KH, Rall, SC Jr & Mahley, RW (1981) Human E apoprotein heterogeneity. Cysteine-arginine interchanges in the amino acid sequence of the apo-E isoforms. Journal of Biological Chemistry 256, 90779083.CrossRefGoogle Scholar
Welty, FK, Lichtenstein, AH, Barrett, PH, Jenner, JL, Dolnikowski, GG & Schaefer, EJ (2000) Effects of ApoE genotype on ApoB-48 and ApoB-100 kinetics with stable isotopes in humans. Arteriosclerosis, Thrombosis, and Vascular Biology 20, 18071810.CrossRefGoogle ScholarPubMed
Wetterau, JR, Aggerbeck, LP, Rall, SC Jr & Weisgraber, KH (1988) Human apolipoprotein E3 in aqueous solution. I. Evidence for two structural domains. Journal of Biological Chemistry 263, 62406248.CrossRefGoogle ScholarPubMed
Wilson, C, Wardell, MR, Weisgraber, KH, Mahley, RW & Agard, DA (1991) Three-dimensional structure of the LDL receptor-binding domain of human apolipoprotein E. Science 252, 18171822.CrossRefGoogle ScholarPubMed
Wilson, PW, Myers, RH, Larson, MG, Ordovas, JM, Wolf, PA & Schaefer, EJ (1994) Apolipoprotein E alleles, dyslipidemia, and coronary heart disease. The Framingham Offspring Study. Journal of the American Medical Association 272, 16661671.CrossRefGoogle ScholarPubMed
Wilson, PW, Schaefer, EJ, Larson, MG & Ordovas, JM (1996) Apolipoprotein E alleles and risk of coronary disease. A meta-analysis. Arteriosclerosis, Thrombosis, and Vascular Biology 16, 12501255.CrossRefGoogle ScholarPubMed
Yue, P, Isley, WL, Harris, WS, Rosipal, S, Akin, CD & Schonfeld, G (2005) Genetic variants of ApoE account for variability of plasma low-density lipoprotein and apolipoprotein B levels in FHBL. Atherosclerosis 178, 107113.CrossRefGoogle ScholarPubMed
Zechner, R, Moser, R, Newman, TC, Fried, SK & Breslow, JL (1991) Apolipoprotein E gene expression in mouse 3T3-L1 adipocytes and human adipose tissue and its regulation by differentiation and lipid content. Journal of Biological Chemistry 266, 1058310588.CrossRefGoogle ScholarPubMed
Zeleny, M, Swertfeger, DK, Weisgraber, KH & Hui, DY (2002) Distinct apolipoprotein E isoform preference for inhibition of smooth muscle cell migration and proliferation. Biochemistry 41, 1182011823.CrossRefGoogle ScholarPubMed
Zhang, B, Liu, ZH, Zeng, CH, Zheng, JM, Chen, HP, Zhou, H & Li, LS (2005) Plasma level and genetic variation of apolipoprotein E in patients with lipoprotein glomerulopathy. Chinese Medical Journal 118, 555560.Google ScholarPubMed
Zhang, WY, Gaynor, PM & Kruth, HS (1996) Apolipoprotein E produced by human monocyte-derived macrophages mediates cholesterol efflux that occurs in the absence of added cholesterol acceptors. Journal of Biological Chemistry 271, 2864128646.CrossRefGoogle ScholarPubMed
Zhao, Y, Thorngate, FE, Weisgraber, KH, Williams, DL & Parks, JS (2005) Apolipoprotein E is the major physiological activator of lecithin-cholesterol acyltransferase (LCAT) on apolipoprotein B lipoproteins. Biochemistry 44, 10131025.CrossRefGoogle ScholarPubMed
Zilversmit, DB (1979) Atherogenesis: a postprandial phenomenon. Circulation 60, 473485.CrossRefGoogle ScholarPubMed
Figure 0

Fig. 1. Key structural elements of apo E (reprinted from Hatters et al.2006, with permission from Elsevier). (a) The amino-terminal domain consists of a four-helix bundle that contains the LDL receptor-binding region of the protein contained between amino acids 136–150 in helix 4. Contained within the ‘hinge region’, amino acid 172 is thought to be essential for receptor binding. The carboxyl-terminal contains the lipoprotein-binding region. (b) The model demonstrates the impact of the replacement of Cys with Arg on position 112 in the protein. This replacement facilitates the interaction between Arg 61 and Glu 255, which mediates closer contact between the amino-terminal and carboxyl-terminal domains.

Figure 1

Table 1. Polymorphisms found in apoE gene exons (data from National Center for Biotechnology Information (2006) single-nucleotide polymorphism database)

Figure 2

Table 2. ApoE isoform amino acid differences and physio-chemical changes

Figure 3

Table 3. ApoE allelic distribution in select populations worldwide (derived from Singh et al.2006)

Figure 4

Table 4. The impact of apoE genotype on LDL-cholesterol levels (E2/E4 excluded if present)

Figure 5

Fig. 2. Local effects of apoE on the artery wall. M, monocyte; MΦ, macrophage; EC, endothelial cell; P, platelet; T, T lymphocyte; SMC, smooth muscle cells; VCAM-1, vascular cell adhesion molecule-1.

Figure 6

Table 5. Proposed roles for apoE in reverse cholesterol transport

Figure 7

Table 6. CHD adjusted hazard ratios (HR) according to apoE genotype for men participating in the Northwick Park Heart Study* (adapted from Humphries et al.2001)