TABLE 1. Logistic Regression Analysis of KL-VS Frequency and Age

Ashkenazi Ashkenazi Ashkenazi (Age Restricted) Czech Combined Ashkenazi (Age Restricted) Czech Combined Age 63 to 81 75 to 81 75 to 81 75 to 81 79 to 104 79 to 94 79 to 94 79 to 94 Beta (cid:5)0.096 (cid:5)0.060 (cid:5)0.0169 (cid:5)0.135 0.052 0.092 0.087 0.089 SE 0.032 0.148 0.100 0.083 0.018 0.034 0.034 0.024 OR 0.908 0.942 0.845 0.874 1.053 1.096 1.091 1.094 95% CI 0.852 to 0.968 0.705 to 1.259 0.695 to 1.028 0.743 to 1.029 1.017 to 1.091 1.026 to 1.171 1.021 to 1.166 1.043 to 1.146 5-year survival OR 0.62 0.74 0.43 0.51 1.29 1.58 1.55 1.57 P 0.003 0.69 0.092 0.11 0.004 0.007 0.010 0.0002

2. Cardiovascular Risk Factors Stratified by KL-VS Genotype

FF (n) FV (n) VV (n) P* Male 33.3% (357) 29.5% (149) 26.3% (19) 0.59 Age, years 85.1(cid:6)11.7 (357) 85.9(cid:6)12.6 (149) 85.3(cid:6)12.5 (19) 0.78 LDL-C, mg/dL 116.9(cid:6)33.2 (309) 110.4(cid:6)32.9 (132) 126.3(cid:6)31.7 (16) 0.07 HDL-C, mg/dL 54.7(cid:6)17.1 (313) 58.3(cid:6)18.1 (136) 50.7(cid:6)14.5 (16) 0.06 Triglycerides, mg/dL 140.1(cid:6)84.8 (321) 140.9(cid:6)103.6 (139) 140.9(cid:6)69.6 (16) 1.0 BMI, kg/m2 23.8(cid:6)3.7 (216) 23.7(cid:6)4.0 (108) 24.7(cid:6)3.7 (12) 0.69 SBP, mm Hg 138.2(cid:6)21.3 (252) 132.0(cid:6)23.4 (122) 153.4(cid:6)39.5 (14) 0.001 DBP, mm Hg 75.3(cid:6)10.9 (252) 73.1(cid:6)12.1 (122) 79.3(cid:6)10.7 (14) 0.07 Diabetes mellitus 17.7% (226) 18.4% (114) 0 (12) 0.27

BMI indicates body mass index; FF, homozygous wild-type; FV, heterozygous; VV, KL-VS.

BMI indicates body mass index; FF, homozygous wild-type; FV, heterozygous; VV, homozygous

*P refers to the significance of differences between all three genotypes as measured by ANOVA for continuous variable and by the (cid:3)2 test for dichotomous variables.

tively; Table 1). When the Ashkenazi cohort is restricted to the same age range as the Czech (79 to 94 years), the heterozygous advantage is still significant (P(cid:1) 0.007), and the Be magnitude of the effect, as indicated by the -estimates, is remarkably similar (0.092(cid:6) 0.034 versus 0.087(cid:6) 0.034). Combining these two data sets, we find a 1.094-fold (95% CI, 1.043 to 1.146) increased odds ratio (OR) for survival per year for heterozygous individuals from age 79 to 94 years in two independent populations. This corresponds to a 1.57-fold (95% CI, 1.23 to 1.98) increased OR for 5-year survival (P(cid:1) 0.0002).

CVD risk factors. Table 2 displays CVD risk factors stratified by KL-VS genotype. Univariate analysis indicates that sys- tolic blood pressure (SBP) levels were significantly associ- ated with KL-VS genotype (P(cid:1) 0.001), and suggestive results (P(cid:1) 0.10) were seen for LDL-C, HDL-C, and DBP. Figure 2 indicates that for each of these measurements, the heterozy- gous individuals have the best CVD risk profile and homozy- gous KL-VS individuals the worst. These data do not fit traditional genetic models, and thus, linear regression was performed for each of these CVD risk factors, with each genotype entered as an independent variable (Table 3). HDL-C was associated significantly with KL-VS genotype after adjusting for potential confounders in a multivariate analysis (P(cid:1) 0.050), with a decrease of 3.47 mg/dL (95% CI,   aut»—0 7.19 to 0.24) and 9.80 mg/dL (95% CI, 19.05 to 0.54) of HDL-C for homozygous wild-type and homozygous KL-VS individuals, respectively, relative to heterozygous individuals. In contrast, KL-VS allele status was not signifi- cantly associated with LDL-C levels after adjustment for potential confounders (P(cid:3) 0.28).

In the Ashkenazi population, a decrease in the frequency of heterozygous individuals between the ages of 63 to 81 years is observed (OR, 0.908; 95% CI, 0.852 to 0.968; P(cid:1) 0.003). Although not noted previously, the Czech population shows a similar decrease in frequency of heterozygous individuals 75 to 81 years of age (OR, 0.845; 95% CI, 0.695 to 1.028; P(cid:1) 0.092). When the Ashkenazi cohort was restricted to the same age range (75 to 81 years), the negative trend for frequency of heterozygous individuals was also not statisti- cally significant (P(cid:1) 0.69), nor was the trend significant in the combined data set (P(cid:1) 0.11).

SBP was significantly associated with KL-VS allele status after adjusting for potential confounders (P(cid:1) 0.008), whereas suggestive data were seen for diastolic blood pressure (DBP; P(cid:1) 0.089; Table 3). Individuals heterozygous for the KL-VS

Analysis of CVD Risk Factors

Previous results have indicated that the KL-VS allele influ- ences the risk for early-onset CAD,5 and therefore, we

LDL (mg/dL); SBP (mm Hg)16015014013012011080mFFmFVawNisHDL-C  HDL (mg/dL); DBP (mm Hg)

TABLE 3. Multivariate Linear Regression Analysis for Cardiovascular Risk Factors*

Risk Factor P † Genotype (cid:2)‡ SE P 95% CI robust   LDL-C, mg/dL 0.28 FF 0.01 4.10 1.0 (cid:5)8.06 to 8.07 VV 14.82 10.18 0.15 (cid:5)5.22 to 34.87 HDL-C, mg/dL 0.050 FF (cid:5)3.47 1.89 0.067 (cid:5)7.19 to 0.24 VV (cid:5)9.80 4.70 0.038 (cid:5)19.05 to (cid:5)0.54 SBP, mm Hg 0.008 FF 6.51 2.82 0.022 0.97 to 12.05 VV 18.97 7.01 0.007 5.19 to 32.77 DBP, mm Hg 0.089 FF 2.57 1.33 0.055 (cid:5)0.05 to 5.19 VV 5.15 3.32 0.12 (cid:5)1.38 to 11.68

*Model is adjusted for age, sex, body mass index, LDL-C, HDL-C, triglycerides, SBP, DBP, and proband status, with the dependent variable excluded from each specific model. SBP and DBP are excluded from the analysis of blood pressure measurements.

†P refers to the overall significance of the KL-VS allele (see Material and Methods); robust ‡homozygous wild-type (FF) and KL-VS (VV) individuals are compared with heterozygous individuals

allele had lower SBP (6.51 mm Hg; 95% CI, 0.97 to 12.05) and DBP (2.57 mm Hg; 95% CI, outs 0.05 to 5.19) than wild-type individuals. Remarkably, homozygosity for the KL-VS allele was associated with an increase of 18.97 mm Hg (95% CI, 5.18 to 32.77) in SBP relative to heterozygous individuals.