Apolipoprotein E deficient rats generated via zinc-finger nucleases exhibit pronounced in-stent restenosis

Abstract

The long-term success of coronary stent implantation is limited by in-stent restenosis (ISR). In spite of a broad variety of animal models available, an ideal high-throughput model of ISR has been lacking. Apolipoprotein E (apoE) deficient rats enable the evaluation of human-sized coronary stents while at the same time providing an atherogenic phenotype. Whereas apoE deficient rats have been proposed as animal model of atherosclerosis, to date it is unknown whether they also develop pronounced ISR. We sought to assess ISR after abdominal aorta stent implantation in apoE deficient rats. A total of 42 rats (16 wildtype, 13 homozygous apoE−/− and 13 heterozygous apoE+/− rats) underwent abdominal aorta stent implantation. After 28 days blood samples were analyzed to characterize lipid profiles. ISR was assessed by histomorphometric means. Homozygous apoE−/− rats exhibited significantly higher total cholesterol and low-density cholesterol levels than wildtype apoE+/+ and heterozygous apoE+/− rats. ISR was significantly pronounced in homozygous apoE−/− rats as compared to wildtype apoE+/+ (p = <0.0001) and heterozygous apoE+/− rats (p = 0.0102) on western diet. Abdominal aorta stenting of apoE−/− rats is a reliable model to investigate ISR after stent implantation and thus can be used for the evaluation of novel stent concepts. Apolipoprotein E (apoE) deficient rats have been proposed as animal model of atherosclerosis. We investigated the development of restenosis 28 days after stent implantation into the abdominal aorta of wildtype apoE+/+, homozygous apoE−/− and heterozygous apoE+/− rats, respectively. Homozygous apoE−/− rats exhibited significantly higher LDL and significantly lower HDL cholesterol levels compared to wildtype apoE+/+ and heterozygous apoE+/− rats. Restenosis after stent implantation was significantly pronounced in western-diet-fed homozygous apoE−/− rats, accompanied by a significantly increased neointimal thickness. Thus, apoE knockout rats exhibit elevated restenosis and might provide a novel tool for testing of innovative stent concepts.

Apolipoprotein E4 Is Associated with Right Ventricular Dysfunction in Dilated Cardiomyopathy—An Animal and In-Human Comparative Study

“…All animal experiments were performed after approval (AZ 84-02.04.2012.A277), according to the European legislation and according to the FELASA guideline for the care and use of experimental animals. Eight-week-old apoE +/+ (Sprague Dawley, Charles River, Cologne, Germany) and apoE −/− (SDapoEtm1sage rats; product number: TGRA3710HM9; Compor Zr ® Zinc-finger nuclease target site: 5′ CAGGCCCTGAACCGAttctggGATTACCTGCGCTGGG; NCBI GeneID: NC_005100.2 GenBank; Sigma Aldrich Genetic Engineering Labs, Saint Louis, MO, USA) [ 8 ] rats were fed a normal diet (3.3% crude fat, cholesterol-free) or western type diet (WD, Ssniff Spezialdiaten, Soest, Germany) for 12 weeks. Rats were kept in standardized conditions (21 °C ± 2 °C, 60% ± 5% humidity, and a 12 h light/dark cycle, free access to food and water ad libitum).…”

Toward the Effective Bioengineering of a Pathological Tissue for Cardiovascular Disease Modeling: Old Strategies and New Frontiers for Prevention, Diagnosis, and Therapy

“…In several animal models, species propensity to spontaneously develop atherosclerosis and/or feeding with cholesterol-rich diets have been exploited to induce or accelerate the development of atherosclerotic lesions in experimental settings. Through transgenesis strategies, it is possible to reproduce more efficiently the human disease phenotype in animal models, classically in mice, rats, rabbits, and in the last years in pigs, too ( 136 , 138 , 139 , 141 ). Of note, hypercholesterolemia is almost the unique factor that has been considered in animal-based approaches ( 138 ).…”

Heart function assessment during aging in apolipoprotein E knock-out mice

“…Apolipoprotein E (apoE) is a ubiquitous protein, essential for the lipid metabolism in the whole body 1,2 . The role of apoE in lipid metabolism, ensuring the fulfillment of cholesterol homeostasis and adequate clearance to circulating lipids 3,4 is the best known. Numerous studies have shown new other functions of this protein, independent of lipid metabolism, thus highlighting the polymorphism and multifunctionality of apoE 5 .…”