Plasma legumain in familial hypercholesterolemia: associations with statin use and cardiovascular risk markers

Figures & data

Figure 1. Plasma levels of legumain in the (A) total population of FH subjects and controls (controls, n = 96; FH, n = 251), (B) in children and young adults (controls, n = 64; FH, n = 85) and (C) in the adult populations (controls, n = 32; FH, n = 166). FH, familial hypercholesterolemia. P values are from Welch Two Sample t-tests.

Figure 2. Plasma levels of legumain in adult FH subjects according to statin use (no statin use, n = 33; statin use, n = 133). FH: familial hypercholesterolemia. p value is from Welch Two Sample t-test.

Table 1. FH subjects and healthy control <18 years of age.

	Overall, N = 149	Controls, N = 64	FH, N = 85	p value
Statin use	0 (0%)	0 (0%)	0 (0%)
Female	70 (47%)	26 (41%)	44 (52%)	0.18
Age, y	13 (10, 14)	14 (13, 14)	12 (10, 14)	<0.001
Weight, kg	48 (15)	52 (13)	46 (16)	0.050
Height, cm	156 (15)	162 (13)	152 (15)	<0.001
BMI, kg/m^2	19.3 (3.3)	19.2 (2.9)	19.3 (3.5)	0.89
TC, mmol/L	5.7 (1.9)	4.0 (0.7)	7.0 (1.5)	<0.001
LDL-C, mmol/L	3.9 (1.8)	2.2 (0.5)	5.1 (1.4)	<0.001
HDL-C, mmol/L	1.4 (0.3)	1.5 (0.3)	1.4 (0.3)	0.027
TG, mmol/L	0.7 (0.5, 1.0)	0.7 (0.5, 1.0)	0.8 (0.6, 1.0)	0.13
ApoB, g/L	1.0 (0.4)	0.6 (0.1)	1.3 (0.3)	<0.001
ApoA-I, g/L	1.4 (0.2)	1.4 (0.2)	1.4 (0.3)	0.67
Glucose, mmol/L	4.78 (0.54)	4.67 (0.62)	4.82 (0.51)	0.33
CRP, mg/L	0.60 (0.60, 1.00)	0.60 (0.60, 0.62)	1.00 (0.60, 1.00)	<0.001
Legumain, pg/mL	2881 (2430, 3,778)	2868 (2423, 3,590)	2910 (2451, 4,005)	0.72

n (%) for categorical variables; median (IQR) and mean (SD) for right-skewed and normally distributed continuous variables, respectively. For hypothesis testing, we used Wilcoxon rank sum test and Welch Two Sample t-test for right-skewed and normally distributed continuous variables, respectively; we used Pearson’s Chi-squared test for categorical variables. FH: familial hypercholesterolemia; BMI: body mass index; TC: total cholesterol; LDL-C: low density lipoprotein cholesterol; HDL-C: high density lipoprotein cholesterol; TG: triglycerides; apoB, apolipoprotein B; apoA-1: apolipoprotein A1; CRP: C reactive protein.

Table 2. FH subjects and healthy control 18 years of age and older.

	Overall, N = 198	Controls, N = 32	FH, N = 166	p value
CHD
No CHD	124 (86%)	0 (0%)	124 (86%)
CHD	21 (14%)	0 (0%)	21 (14%)
Statin use				<0.001
No statin use	65 (33%)	32 (100%)	33 (20%)
Statin use	133 (67%)	0 (0%)	133 (80%)
Sex				0.15
Male	91 (46%)	11 (34%)	80 (48%)
Female	107 (54%)	21 (66%)	86 (52%)
Age, y	40 (27, 56)	29 (24, 41)	42 (28, 58)	0.001
Weight, kg	76 (17)	67 (13)	77 (17)	<0.001
Height, cm	173 (9)	174 (10)	173 (9)	0.39
BMI, kg/m^2	25.2 (5.0)	21.7 (2.8)	25.9 (5.0)	<0.001
TC, mmol/L	5.1 (1.8)	4.5 (0.7)	5.3 (2.0)	<0.001
LDL-C, mmol/L	3.3 (1.8)	2.6 (0.7)	3.4 (1.9)	<0.001
HDL-C, mmol/L	1.5 (0.4)	1.6 (0.5)	1.4 (0.4)	0.11
TG, mmol/L	1.0 (0.7, 1.5)	0.9 (0.7, 1.3)	1.0 (0.7, 1.5)	0.21
ApoB, g/L	1.0 (0.4)	0.8 (0.2)	1.1 (0.4)	<0.001
ApoA-I, g/L	1.5 (0.3)	1.6 (0.3)	1.5 (0.3)	0.049
Glucose, mmol/L	5.40 (0.94)	4.97 (1.10)	5.45 (0.91)	0.12
CRP, mg/L	0.60 (0.60, 1.08)	0.65 (0.60, 1.23)	0.60 (0.60, 0.99)	0.47
Legumain, pg/mL	5264 (4061, 6,707)	4003 (3435, 5,120)	5560 (4465, 6,735)	<0.001

n (%) for categorical variables; median (IQR) and mean (SD) for right-skewed and normally distributed continuous variables, respectively. For hypothesis testing, we used Wilcoxon rank sum test and Welch Two Sample t-test for right-skewed and normally distributed continuous variables, respectively; we used Pearson’s Chi-squared test for categorical variables. FH: familial hypercholesterolemia; CHD: coronary heart disease; BMI: body mass index; TC: total cholesterol; LDL-C: low density lipoprotein cholesterol; HDL-C: high density lipoprotein cholesterol; TG: triglycerides; apoB: apolipoprotein B; apoA-1: apolipoprotein A1; CRP: C reactive protein.

Figure 3. Association between legumain and clinical and inflammatory/platelet activation markers in the total population of FH subjects and controls (right), only FH subjects (Middle) and controls (left). The volcano plots show regression coefficients and 95% CIs from multiple regression analyses adjusted for age, sex, BMI, and statin use (statin use not relevant for control group). Colors indicate p-value. Prior to modeling, to enable visualization in a volcano plot, we scaled each clinical and inflammatory/platelet activation marker to a standard normal distribution by subtracting the mean and dividing by its standard deviation. FH: familial hypercholesterolemia; LDL-C: low-density lipoprotein cholesterol; HDL-C: high-density lipoprotein cholesterol; apoB: apolipoprotein B; apoA-1: apolipoprotein A1; CRP: C reactive protein; Lp-PLA2: LDL-associated phospholipase A2; PF4: platelet factor 4; NAP2: neutrophil activating protein; TNF: tumor necrosis factor; SD: standard deviation.