Overexpression of tissue-nonspecific alkaline phosphatase (TNAP) in endothelial cells accelerates coronary artery disease in a mouse model of familial hypercholesterolemia.

Filippo Romanelli,Alexei Y Savinov,A Martin Gerdes,Anthony B Pinkerton,Anthonymarco Corbo,Manisha C Yadav,Soha Salman,Maria Plummer,David Petrosian,José Luis Millán,Olga V Savinova,Kenneth B Margulies,Maryam Salehi,Jes Kuruvilla,Ilian Radichev,Jesse Chait,Omid J Rashidbaigi,Elena Aikawa

doi:10.1371/journal.pone.0186426

Abstract

ObjectiveOverexpression of tissue-nonspecific alkaline phosphatase (TNAP) in endothelium leads to arterial calcification in mice. The purpose of this study was to examine the effect of elevated endothelial TNAP on coronary atherosclerosis. In addition, we aimed to examine endogenous TNAP activity in human myocardium.Approach and resultsA vascular pattern of TNAP activity was observed in human non-failing, ischemic, and idiopathic dilated hearts (5 per group); no differences were noted between groups in this study. Endothelial overexpression of TNAP was achieved in mice harboring a homozygous recessive mutation in the low density lipoprotein receptor (whc allele) utilizing a Tie2-cre recombinase (WHC-eTNAP mice). WHC-eTNAP developed significant coronary artery calcification at baseline compared WHC controls (4312 vs 0μm2 alizarin red area, p<0.001). Eight weeks after induction of atherosclerosis, lipid deposition in the coronary arteries of WHC-eTNAP was increased compared to WHC controls (121633 vs 9330μm2 oil red O area, p<0.05). Coronary lesions in WHC-eTNAP mice exhibited intimal thickening, calcifications, foam cells, and necrotic cores. This was accompanied by the reduction in body weight and left ventricular ejection fraction (19.5 vs. 23.6g, p<0.01; 35% vs. 47%, p<0.05). In a placebo-controlled experiment under atherogenic conditions, pharmacological inhibition of TNAP in WHC-eTNAP mice by a specific inhibitor SBI-425 (30mg*kg-1*d-1, for 5 weeks) reduced coronary calcium (78838 vs.144622μm2) and lipids (30754 vs. 77317μm2); improved body weight (22.4 vs.18.8g) and ejection fraction (59 vs. 47%). The effects of SBI-425 were significant in the direct comparisons with placebo but disappeared after TNAP-negative placebo-treated group was included in the models as healthy controls.ConclusionsEndogenous TNAP activity is present in human cardiac tissues. TNAP overexpression in vascular endothelium in mice leads to an unusual course of coronary atherosclerosis, in which calcification precedes lipid deposition. The prevalence and significance of this mechanism in human atherosclerosis requires further investigations.

Highlights

Coronary artery calcification is an independent predictor of cardiovascular morbidity and mortality [1,2,3,4,5,6,7,8]
Endogenous Tissue-nonspecific alkaline phosphatase (TNAP) activity is present in human cardiac tissues
TNAP overexpression in vascular endothelium in mice leads to an unusual course of coronary atherosclerosis, in which calcification precedes lipid deposition

Summary

Introduction

Coronary artery calcification is an independent predictor of cardiovascular morbidity and mortality [1,2,3,4,5,6,7,8]. Tissue-nonspecific alkaline phosphatase (TNAP) promotes vascular calcification by regulating the levels of pyrophosphate (PPi), an inhibitor of mineralization. Upregulation of TNAP expression in vascular smooth muscle cells or in endothelial cells leads to pathological calcification in animal models [10, 11]. Increased TNAP expression has been detected in patients suffering from vascular calcification associated with diabetes [12] and chronic kidney disease [13, 14]. Recent epidemiological studies point to a potential pathophysiological role and a prognostic value of increased blood alkaline phosphatase activity in human coronary artery disease [15, 16]

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