Association of Circulating Endothelial Microparticles with Cardiovascular Risk Factors in the Framingham Heart Study

N. Amabile, S. Cheng, JM. Renard, MG. Larson, A. Ghorbani,

E. McCabe, G. Griffin, RS. Vasan, SY. Shaw, KS. Cohen,

C. Guerin, A. Tedgui, CM. Boulanger, TJ. Wang

Disclosure

• No disclosure

Background

• Endothelial microparticles (EMPs) are released during endothelial cell apoptosis and/or injury.

• EMPs interfere with vascular inflammation, coagulation pathways, and regulation of vascular tone.

Background: EMPs and CV risk factors

Preston R A et al. Hypertension 2003;41:211-217

Control Mild Severe

Hypertension

EM

Ps

(m

illi

on

/ml)

Diabetes

Sabatier F et al. 2002;51(9):2480-85

EM

Ps /µ

L

Background

• Clinical studies in select patient samples have demonstrated that circulating EMPs levels are increased in the presence of cardiovascular risk factors such as diabetes, dyslipidemia, and uncontrolled systemic hypertension.

• Because data on EMPs in humans has been limited to referral samples, the extent to which circulating EMPs may reflect chronic endothelial injury and cardiovascular risk in the general population is unknown

Aims

• We investigated whether high EMPs concentrations were associated with the presence of cardiovascular and metabolic risk factors in a large community-based population.

Methods

• N=844 individuals without history of cardiovascular disease from the Framingham Heart Study Offspring cohort were included in this study.

• All participants underwent a standardized assessment of cardiovascular risk factors, clinical examination and a fasting blood sample was drawn.

• Glucose, triglycerides and total and HDL Cholesterol values were measured.

• Metabolic syndrome was defined according to the NCEP criteria.

Methods (2)

• Poor platelet plasma was prepared by successive centrifugation steps.

• Endothelial microparticles (EMPs) levels were measured using flow cytometry methods on a Guava EasyCyte 8HT device.

• 3 subpopulations of EMPs were analyzed: CD144+ (VE-Cadherin) EMPs

CD62e+ (E selectin) EMPs

CD31+ (PECAM)/CD41- EMPs

Baseline characteristics (N=844)

Age, y 669

Female, % 57

Systolic blood pressure, mm Hg 12717

Diastolic blood pressure, mm Hg 7410

Body mass index, kg/m2 28.15.1

Waist circumference, inches 39.95.6

Cigarette smoking, % 8

Total cholesterol, mg/dL 18636

HDL cholesterol, mg/dL 5918

Total/HDL cholesterol ratio 3.41.0

Triglycerides, mg/dL 10955

Hypertension, % 59

Diabetes mellitus, % 12

Anti-hypertensive therapy, % 48

Cholesterol lowering therapy, % 46

Metabolic syndrome, % 48

Framingham Risk Score 8.53.9

CD144+ EMPs CD62e+ EMPs CD31+ EMPs

Regression Coefficient

p Regression Coefficient

p Regression Coefficient

p

Age, year -0.032 (0.029) 0.15 0.037 (0.029) 0.19 0.020 (0.032) 0.52

Female sex -0.113 (0.056) 0.045 -0.139 (0.055) 0.012 -0.055 (0.062) 0.37

Body mass index, kg/m2 0.066 (0.028) 0.020 0.005 (0.028) 0.87 0.062 (0.031) 0.043

Waist circumference, inch. 0.088 (0.028) 0.002 0.021 (0.028) 0.45 0.093 (0.031) 0.003

Systolic BP, mm Hg 0.044 (0.031) 0.15 0.018 (0.030) 0.56 0.051 (0.033) 0.13

Diastolic BP, mm Hg 0.068 (0.030) 0.024 0.026 (0.029) 0.37 0.064 (0.033) 0.050

Hypertension 0.202 (0.059) 0.0007 0.175 (0.058) 0.003 0.064 (0.066) 0.33

Total/HDL chol. ratio 0.112 (0.029) <0.0001 0.042 (0.028) 0.14 0.176 (0.031) <0.0001

Total chol., mg/dL 0.001 (0.001) 0.16 0.000 (0.001) 0.55 0.001 (0.001) 0.31

HDL chol., mg/dL -0.005 (0.002) 0.005 -0.001(0.002) 0.74 -0.009 (0.002) <0.0001

Log triglycerides 0.149 (0.038) <0.0001 0.029 (0.027) 0.29 0.226 (0.030) <0.0001

Diabetes 0.048 (0.088) 0.59 -0.004 (0.085) 0.96 0.006 (0.096) 0.95

Current smoker -0.045 (0.105) 0.67 -0.169 (0.103) 0.10 0.044 (0.116) 0.70

Age- and Sex-Adjusted relations of clinical characteristics with EMPs

CD144+ EMPs CD62e+ EMPs CD31+ EMPs

Regression Coefficient

p Regression Coefficient

p Regression Coefficient

p

Age, year -0.025 (0.018) 0.16 0.007 (0.016) 0.67 0.016 (0.030) 0.59

Female sex -0.043 (0.035) 0.22 -0.058 (0.032) 0.07 -0.019 (0.058) 0.74

Waist circumference, inch. 0.020 (0.018) 0.26 -0.006 (0.016) 0.69 0.041 (0.030) 0.17

Hypertension 0.083 (0.037) 0.025 0.094 (0.034) 0.005 -0.014 (0.061) 0.81

Diabetes -0.044 (0.053) 0.41 -0.025 (0.048) 0.60 -0.111 (0.087) 0.20

Total/HDL chol. ratio 0.018 (0.022) 0.40 0.026 (0.020) 0.20 0.039 (0.036) 0.27

Log triglycerides 0.068 (0.022) 0.002 -0.005 (0.020) 0.80 0.166 (0.036) <0.0001

Multivariable-adjusted relations of clinical characteristics with EMPs phenotypes

CD144+ EMPs CD62e+ EMPs CD31+ EMPs

Regression Coefficient

p Regression Coefficient

p Regression Coefficient

p

Age, year -0.06 (0.03) 0.04 0.03 (0.03) 0.23 0.01 (0.03) 0.7

Female sex -0.15 (0.06) 0.02 -0.13 (0.06) 0.03 -0.14 (0.06) 0.02

Metabolic syndrome 0.25 (0.06) <0.001 0.13 (0.06) 0.04 0.10 (0.06) 0.1

Age- and Sex-Adjusted relations of Metabolic syndrome with EMPs in patients without diabetes (n=741)

FRS by tertiles of circulating CD31+ EMPs

1st Tertile 2nd Tertile 3rd Tertile

p=0.03

Discussion

• These data indicates that EMPs levels are associated with several cardiovascular risk factors in a large community-based cross sectional analysis.

• EMPs levels are related to TG levels and presence of metabolic syndrome, which confirms the deleterious impact of these abnormalities on endothelium.

• As EMPs display potential pro-coagulant/pro-inflammatory properties, they might represent mediators enhancing the deleterious impact of high TG and MS on cardiovascular homeostasis

TG-rich LPs

HYPERINSULINEMIA

HYPERGLYCEMIA

ADIPOCYTOKINES

Adipocytes

Monocytes END. CELL

Inflammation Oxidative Stress

END DYSFUNCTION

HYPERTENSION

Pro Inflammatory

Pro Coagulant

Conclusion

• These data underscores the potential influence of high-risk metabolic profiles on endothelial integrity.

• Our results advocate for the need of further investigation to explore the potential mechanisms linking metabolic and lipids abnormalities to production of endothelial MPs

Effect of 80 mg atorvastatin treatment on circulating CD144+ EMPs levels

N=19 patients with symptomatic PAD ; 8 weeks of atorvastatin treatment

Mobarrez F.. et al , Thrombosis Research 2012; 129 (1) :95-97

Effect of 10/40 mg atorvastatin treatment on circulating CD31+ EMPs levels

0

500

1000

1500

2000

2500

Control group(n=100)

Ator 10 mg group(n=48)

Ator 40 mg group(n=48)

Baseline

12 months

*

Huang B. et al , Clinical Cardiology 2012; 35 (2) :125-30