Introduction
Materials and methods
Study participants
Evaluation of cardiovascular outcomes
Isolation of HDL2 and HDL3 from serum
Measurement of serum amyloid A
Genotyping
Statistical analysis
Results
Subject characteristics
Control (n = 150) | CVD (n = 102) | p value | |
---|---|---|---|
Age (years) | 65 (10) | 66 (9) | 0.52 |
Males, n (%) | 83 (55) | 60 (59) | 0.34 |
Diabetes, number with diabetes (%) | 55 (37) | 40 (39) | 0.39 |
DBP (mmHg) | 81 (81) | 81 (81) | 0.76 |
SBP (mmHg) | 132 (118, 148) | 135 (123, 151) | 0.31 |
MABP (mmHg) | 98 (90, 106) | 98 (90, 108) | 0.75 |
HDL cholesterol (mmol/L) | 1.3 (1.0, 1.6) | 1.2 (1.0, 1.5) | 0.64 |
LDL cholesterol (mmol/L) | 2.2 (1.7, 2.9) | 2.0 (1.5, 2.6) | 0.07 |
Total cholesterol (mmol/L) | 3.8 (3.0, 4.8) | 3.6 (3.1, 4.3) | 0.10 |
Troponin (ng/L) | 9.9 (5.0, 16.0) | 10.5 (5.8, 15.7) | 0.72 |
HbA1c (mmol/mol) | 49 (39, 62) | 51(38, 69) | 0.35 |
CRP (mg/L) | 2.91 (1.22, 5.35) | 2.82 (0.95, 4.70) | 0.86 |
eGFR (mL/min/1.73 m2) | 97(75, 116) | 95 (71, 110) | 0.70 |
Weight (kg) | 87 (78, 100) | 87 (76, 102) | 0.84 |
Demi span (cm) | 80 (80) | 81 (81) | 0.30 |
Anti-platelet, n (%) | 46 (38) | 56 (55) | < 0.001* |
Anti-coagulant, n (%) | 27 (19) | 20 (20) | 0.49 |
Anti-oestrogen, n (%) | 1 (0.7) | 1 (1.0) | 0.65 |
Anti-convulsants, n (%) | 9 (6.2) | 11 (11) | 0.13 |
Anti-hypertensives, n (%) | 53 (37) | 32 (32) | 0.26 |
Hypoglycaemics, n (%) | 38 (26) | 32 (32) | 0.21 |
Statins, n (%) | 95 (66) | 78 (77) | 0.04* |
Corticosteroids, n (%) | 28 (19) | 15 (15) | 0.24 |
Beta-blockers, n (%) | 43 (30) | 49 (49) | < 0.001* |
ACE inhibitors, n (%) | 38 (26) | 42 (42) | < 0.001* |
Calcium channel blocker, n (%) | 33 (22.8) | 23 (22.5) | 0.55 |
Nitrates, n (%) | 4 (2.8) | 27 (26.7) | < 0.001* |
Diuretics, n (%) | 36 (24.8) | 29 (28.7) | 0.30 |
NSAIDs, n (%) | 7 (4.9) | 6 (6.0) | 0.46 |
Preliminary screening
Variants associated with serum amyloid A levels
Exon | SNP | Minor allele | MAF | βeta (95% CI) | p value |
---|---|---|---|---|---|
SAA1 exon 1
| rs11024595 | T | 0.26 | 0.73 (0.61–1.01) | 0.06 |
rs2445166 | A | 0.10 | 0.72 (0.51–1.00) | 0.09 | |
rs11545466 | G | 0.07 | 0.83 (0.56–1.18) | 0.27 | |
rs549103464 | Insertion | 0.07 | 1.07 (0.87–1.25) | 0.65 | |
SAA1 exon 3
| rs11024597 | T | 0.34 | 0.97 (0.81–1.19) | 0.83 |
rs149926073 | A | < 0.01 | 3.37 (1.70–4.00) | 0.02* | |
rs1136743 | C | 0.32 | 3.55 (0.89–1.32) | 0.40 | |
rs11545468 | C | 0.08 | 0.82 (0.55–1.13) | 0.20 | |
rs1136745 | C | 0.16 | 0.70 (0.53–0.94) | 0.02* | |
rs1136747 | T | 0.24 | 1.53 (1.11–1.73) | 0.01* | |
SAA1 exon 4
| rs15790 | T | 0.08 | 1.28 (0.94–1.80) | 0.11 |
rs145680768 | C | < 0.01 | 0.95 (0.31–2.33) | 0.75 | |
rs12218 | T | 0.46 | 1.22 (0.93–1.41) | 0.19 | |
rs1059571 | G | < 0.01 | 0.73 (0.19–0.97) | 0.32 | |
SAA2 exon 4
| rs11540206 | A | < 0.01 | 1.14 (0.61–3.79) | 0.37 |
rs149402852 | A | < 0.01 | 0.83 (0.28–1.33) | 0.21 | |
rs116861605 | A | 0.01 | 2.69 (0.68–2.16) | 0.51 | |
rs2468844 | G | 0.18 | 1.73 (1.14–1.75) | 0.01* |
SNP | Beta (95% CI)1 | p value1 | Beta (95% CI)2 | p value 2 |
---|---|---|---|---|
rs1136745 | 0.65 (0.48–0.24) | 0.01 | 0.72 (0.54–0.74) | 0.03 |
rs2468844 | 1.43 (1.13–1.79) | < 0.001 | 1.32 (1.55–1.67) | 0.02 |
rs149926073 | 1.53 (0.70–3.37) | 0.29 | 2.66 (1.11–6.33) | 0.03 |
rs1136747 | 1.43 (1.13–1.80) | < 0.001 | 1.30 (1.35–1.65) | 0.03 |
Associations of genetic variants with cardiovascular disease status
SNP | Minor allele | Controls, n (%) | CVD, n (%) | Odds ratio (95% CI) | p value |
---|---|---|---|---|---|
rs11024595 | T | 49 (17) | 33 (16) | 0.94 (0.59–1.63) | 0.98 |
rs2445166 | A | 26 (9) | 15 (7) | 0.80 (0.40–1.61) | 0.54 |
rs11545466 | G | 18 (6) | 11 (5) | 0.89 (0.42–1.89) | 0.74 |
rs549103464 | Insertion | 18 (6) | 12 (6) | 1.00 (0.70–1.43) | 0.99 |
rs11024597 | T | 99 (33) | 61 (30) | 0.95 (0.65–1.38) | 0.78 |
rs1136743 | C | 94 (31) | 59 (29) | 0.94 (0.64–1.39) | 0.76 |
rs11545468 | C | 19(6) | 19 (9) | 1.71 (0.85–3.44) | 0.13 |
rs1136745 | C | 42 (14) | 31 (15) | 1.23 (0.70–2.16) | 0.47 |
rs15790 | T | 22 (7) | 16 (8) | 1.03 (0.54–2.00) | 0.93 |
rs145680768 | C | 4 (1) | 0 (0) | 0.00 (0.00) | 1.00 |
rs12218 | T | 122 (41) | 88 (43) | 1.06 (0.71–1.60) | 0.77 |
rs1059571 | G | 6 (2) | 0 (0) | 0.00 (0.00) | 1.00 |
rs112509629 | T | 0 (0) | 5 (2) | 0.00 (0.00) | 1.00 |
rs11540206 | A | 1 (0.3) | 4 (2) | 6.07 (0.67–55.1) | 0.11 |
rs149402852 | A | 4 (1) | 3 (1) | 1.10 (0.24–5.03) | 0.90 |
rs116861605 | A | 7 (2) | 6 (3) | 1.27 (0.41–3.91) | 0.68 |
rs2468844 | G | 60 (20) | 27 (13) | 0.65 (0.41–1.04) | 0.07 |
rs149926073 | A | 4 (1) | 3 (1) | 1.15 (0.51–5.26) | 0.86 |
rs1136747 | T | 68 (23) | 41 (20) | 0.89 (0.57–1.40) | 0.63 |
Serum amyloid A 1 haplotypes
Haplotype | n (%) | Mean serum SAA, mg/L (IQR) | Beta (95% CI)1 | p value1 | Beta (95% CI)2 | p value2 |
---|---|---|---|---|---|---|
1.1/1.1 | 109 (46) | 21 (11, 45) | Reference | Reference | ||
1.1/1.3 | 19 (8) | 16 (3.7, 73) | 0.74 (0.45–1.21) | 0.23 | 0.80 (0.48–1.33) | 0.40 |
1.1/1.5 | 80 (34) | 26 (15, 70) | 1.25 (0.93–1.67) | 0.14 | 1.32 (0.79–1.79) | 0.07 |
1.3/1.3 | 9 (4) | 9.1 (4.9, 19) | 0.43 (0.22–0.85) | 0.02* | 0.34 (0.17–0.69) | < 0.001* |
1.3/1.5 | 7 (3) | 55 (17, 87) | 2.58 (1.19–5.57) | 0.02* | 2.48 (1.16–5.32) | 0.02* |
1.5/1.5 | 11 (5) | 32 (11, 72) | 1.51 (0.81–2.82) | 0.19 | 1.50 (0.79–2.83) | 0.21 |
Haplotype | n (%) | Mean SAA mg/L (IQR) | Beta estimate (95% CI)1 | p value1 | Beta estimate (95% CI)2 | p value2 |
---|---|---|---|---|---|---|
HDL2SAA | ||||||
1.1/1.1 | 109 (46) | 0.70 (0.26, 2.50) | Reference | Reference | ||
1.1/1.3 | 19 (8) | 0.60 (0.21, 1.58) | 0.91 (0.75, 1.20) | 0.66 | 0.79 (0.60, 1.17) | 0.80 |
1.1/1.5 | 80 (34) | 0.80 (0.31, 2.50) | 1.12 (0.70, 2.05) | 0.50 | 1.23 (0.77, 2.54) | 0.27 |
1.3/1.3 | 9 (4) | 0.25 (0.11, 0.57) | 0.64 (0.33, 0.97) | 0.04* | 0.56 (0.26, 0.88) | 0.02* |
1.3/1.5 | 7 (3) | 4.06 (2.78, 6.00) | 1.94 (1.46, 5.10) | < 0.001* | 2.03 (1.47, 6.27) | < 0.001* |
1.5/1.5 | 11 (5) | 0.60 (0.27, 1.61) | 0.93 (0.31, 2.29) | 0.74 | 0.84 (0.22, 2.09) | 0.49 |
HDL3SAA | ||||||
1.1/1.1 | 109 (46) | 10 (5.0, 24) | Reference | Reference | ||
1.1/1.3 | 19 (8) | 10 (3.0, 33) | 0.99 (0.73, 1.36) | 0.96 | 1.06 (0.74, 1.49) | 0.79 |
1.1/1.5 | 80 (34) | 12 (6.0, 29) | 1.17 (0.69, 2.87) | 0.34 | 1.25 (0.74, 3.53) | 0.23 |
1.3/1.3 | 9 (4) | 4.0 (2.0, 7.0) | 0.58 (0.18, 0.79) | 0.01* | 0.48 (0.12, 0.60) | < 0.001* |
1.3/1.5 | 7 (3) | 43 (26, 60) | 2.11 (2.04, 11.1) | < 0.001* | 2.15 (2.05, 14.7) | < 0.001* |
1.5/1.5 | 11 (5) | 0.7 (0.3, 2.5) | 1.35 (0.69, 10.3) | 0.16 | 1.34 (0.56, 12.3) | 0.22 |
Genotype | Controls, n (%) | Cases, n (%) | Odds ratio (95% CI)1 | p value1 | Odds ratio (95% CI)2 | p value2 |
---|---|---|---|---|---|---|
1.1/1.1 | 64 (45) | 45 (48) | Reference | Reference | ||
1.1/1.3 | 12 (9) | 7 (8) | 0.83 (0.30–2.27) | 0.72 | 0.87 (0.28–2.73) | 0.82 |
1.1/1.5 | 50 (35) | 30 (32) | 0.53 (0.47–1.54) | 0.60 | 0.90 (0.46–1.75) | 0.76 |
1.3/1.3 | 4 (3) | 5 (5) | 1.78 (0.45–6.99) | 0.41 | 1.72 (0.34–8.67) | 0.51 |
1.3/1.5 | 6 (4) | 1 (1) | 0.24 (0.03–2.04) | 0.19 | 0.23 (0.02–2.26) | 0.21 |
1.5/1.5 | 6 (4) | 5 (5) | 1.19 (0.34–4.12) | 0.80 | 0.96 (0.23–3.91) | 0.96 |
1.1 allele | 190 (63) | 127 (68) | Reference | Reference | ||
1.3 allele | 26 (18) | 18 (10) | 0.97 (0.51–1.83) | 0.97 | 0.82 | 0.93 |
1.5 allele | 68 (48) | 41 (22) | 1.11 (0.71–1.74) | 0.65 | 0.89 | 0.48 |