Effect of Apolipoprotein E Alleles and Angiotensin- Converting Enzyme Insertion/Deletion Polymorphisms on Lipid and Lipoprotein Markers in Middle-Aged Men and in Patients With Stable Angina Pectoris or Healed Myocardial Infarction Pedro Marques-Vidal, MD, PhD, Vanina Bongard, MD, Jean-Bernard Ruidavets, MD, Josette Fauvel, PhD, Bertrand Perret, MD, PhD, and Jean Ferrie `res, MD The effects of the apolipoprotein E epsilon and angio- tensin-converting enzyme insertion/deletion alleles on lipid levels and hypolipidemic drug treatment was assessed in 400 men with stable angina pectoris or healed myocardial infarction and 338 healthy con- trols. The data indicate that 4 carriers have in- creased total and low-density lipoprotein cholesterol levels, that the 4 allele unfavorably decreases the efficiency of statin treatment, and that the angioten- sin-converting enzyme insertion/deletion polymor- phism exerts no significant effect, with the exception of an increase in apolipoprotein E levels. 2003 by Excerpta Medica, Inc. (Am J Cardiol 2003;92:1102–1105) W e used the data from a large case-control study conducted in southwestern France to assess the effects of apolipoprotein (apo) E alleles and the an- giotensin-converting enzyme insertion/deletion (I/D) polymorphism on lipid and lipoprotein levels in sub- jects treated for dyslipidemia. ••• The methods of the case-control study have been presented previously. 1 Patients with coronary artery disease (CAD) were men aged 35 to 64 years (mean 55) with previous myocardial infarction at least 6 months after the event or stable angina pectoris; pa- tients were consecutively referred to our university hospital (Toulouse, France) for coronary angiography over a 24-month period. The control subjects were healthy men (also aged 35 to 64 years) recruited during the same period; controls were randomly drawn from the electoral rolls of the Toulouse region. Only subjects without clinical evidence of CAD ac- cording to a physical examination, personal and fam- ily history, or an electrocardiogram were included in the control group. Cardiovascular risk factors, drug treatments, and personal history of CAD were assessed by question- naire in all subjects. Professional activity was based on current status (employed vs unemployed/retired/ sick leave). Educational level was based on the num- ber of years spent in school or at a university. Height and weight were measured using common scales. Body mass index was calculated as weight (kilo- grams)/height (meters squared). Overweight was de- fined as a body mass index 25 but 30 kg/m 2 ; obesity was defined as a body mass index 30 kg/m 2 . Systolic and diastolic blood pressures were measured twice on the right arm of subjects who were seated for at least 5 minutes in a comfortable position. Two consecutive measurements were recorded and the mean values were used in this analysis. 2 DNA was obtained from leukocytes using the salt- ing out procedure, 3 and apo E genotyping was per- formed in duplicate for each subject using a polymer- ase chain reaction as described previously. 4 Subjects were classified into 3 categories: 2 carriers (E2 ho- mozygotes and E2/E3 carriers), 3 homozygotes, and 4 carriers (E4 homozygotes and E3/E4 carriers). Ep- silon2/4 carriers (17 subjects) were excluded from the analysis. The presence (allele I) or absence (allele D) of the 287-bp Alu repeat in intron 16 of the angiotensin- converting enzyme gene was assessed by evaluating the size of DNA fragments after polymerase chain reaction amplification as described by Rigat et al. 5 Because some ID genotypes may be misclassified as DD, all DD genotypes were further subjected to a second polymerase chain reaction with insertion-spe- cific primers. 1 Plasma prepared with ethylene di-amino tetra ace- tate was used for the analysis of lipids and lipopro- teins. Plasma total cholesterol and triglycerides were assessed by enzymatic methods (Dimension; Dade- Behring, Le Pont de Claix, France). High-density li- poprotein (HDL) cholesterol was assessed after apo B-containing lipoprotein precipitation using phospho- tungstate-magnesium 2+ ; low-density lipoprotein (LDL) cholesterol was calculated using Friedewald’s formula. Apos A-I and B and lipoprotein(a) [Lp(a)] were measured by immunoturbidimetry in an auto- mated analyzer (Cobas-Mira; Roche Diagnostics, Penz- berg, Germany). Apo E and lipoparticles LpA-I, LpB:E, and LpB:C-III were assayed by a combination of immunoelectrodiffusion and immunoprecipitation techniques (Sebia, Issy-les-Moulineaux, France). The From INSERM U558, Faculte ´ de Me ´ decine Purpan; and INSERM U563, De ´ partement Lipoprote ´ines et Me ´ diateurs Lipidiques, CHU Purpan, Toulouse, France. This report was supported by grants from the Fondation de France Paris; and Bayer Pharmaceuticals, Puteaux, France. Dr. Ferrie ` res’ address is: INSERM U558, Faculte ´ de Me ´- decine, De ´ partement d’Epide ´miologie, 1er e ´tage, 37, Alle ´es Jules Guesde, 31073 Toulouse Cedex, France. E-mail: ferriere@cict.fr. Manuscript received March 5, 2003; revised manuscript received and accepted June 27, 2003. 1102 ©2003 by Excerpta Medica, Inc. All rights reserved. 0002-9149/03/$–see front matter The American Journal of Cardiology Vol. 92 November 1, 2003 doi:10.1016/S0002-9149(03)01046-4