13 research outputs found

    Application of zebrafish and murine models in lipoprotein metabolism and atherosclerosis research

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    Cardiovascular diseases are still a major concern for the global health. The main underlying pathology of this disease is atherosclerosis which is characterized by the accumulation of lipids and immune cells in the arterial wall leading to a chronic local inflammation and lesion formation. In this thesis, we aimed to (1) validate the use of zebrafish in cholesterol metabolism and atherosclerosis research, (2) study the role of certain classes of scavenger receptors in lipoprotein uptake and cholesterol-based functions, and (3) validated two immune-based potential targets for atherosclerosis.Biopharmaceutic

    Zebrafish as outgroup model to study evolution of scavenger receptor class B type I functions

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    Background and aims: Scavenger receptor class B1 (SCARB1) -also known as the high-density lipoprotein (HDL) receptor -is a multi-ligand scavenger receptor that is primarily expressed in liver and steroidogenic organs. This receptor is known for its function in reverse cholesterol transport (RCT) in mammals and hence disruption leads to a massive increase in HDL cholesterol in these species. The extracellular domain of SCARB1 -which is important for cholesterol handling -is highly conserved across multiple vertebrates, except in zebrafish. Methods: To examine the functional conservation of SCARB1 among vertebrates, two stable scarb1 knockout zebrafish lines, scarb1 715delA (scarb1-1 nt) and scarb1 715_716insGG (scarb1 +2 nt), were created using CRISPR-Cas9 technology. Results: We demonstrate that, in zebrafish, SCARB1 deficiency leads to disruption of carotenoid-based pigmen-tation, reduced fertility, and a decreased larvae survival rate, whereas steroidogenesis was unaltered. The observed reduced fertility is driven by defects in female fertility (-50 %, p Drug Delivery Technolog

    PRMT4 inhibitor TP-064 impacts both inflammatory and metabolic processes without changing the susceptibility for early atherosclerotic lesions in male apolipoprotein E knockout mice

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    Atherosclerotic cardiovascular disease is a metabolic and inflammatory disorder. In vitro studies have suggested that protein arginine methyltransferase 4 (PRMT4) may act as a transcriptional coactivator to modulate inflammatory and metabolic processes. Here we investigated the potential anti-atherogenic effect of PRMT4 inhibitor TP-064 in vivo.\nMale apolipoprotein E knockout mice fed a high cholesterol/high fat Western-type diet were intraperitoneally injected three times a week with 2.5 mg/kg (low dose) or 10 mg/kg (high dose) TP-064 or with DMSO control.\nTP-064 induced a dose-dependent decrease in lipopolysaccharide-induced ex vivo blood monocyte Tnfα secretion (p Biopharmaceutic

    Stabilin 1 and 2 are important regulators for cellular uptake of apolipoprotein B-containing lipoproteins in zebrafish

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    BACKGROUND AND AIMS\nMETHODS\nRESULTS\nCONCLUSIONS\nScavenger receptors form a superfamily of membrane-bound receptors that bind and internalize different types of ligands, including pro-atherogenic oxidized low-density lipoproteins (oxLDLs). In vitro studies have indicated a role for the liver sinusoidal endothelial cell receptors stabilin 1 (stab1) and 2 (stab2) in oxLDL clearance. In this study, we evaluated the potential role of stab1 and stab2 in lipoprotein uptake in zebrafish, an upcoming model for studying cholesterol metabolism and atherosclerosis.\nLipoproteins were injected in the duct of Cuvier of wild-type (ABTL) or stab1 and stab2 mutant (stab1-/-stab2-/-) zebrafish larvae at 3 days post-fertilization. To examine the effect of stabilin deficiency on lipoprotein and cholesterol metabolism, zebrafish larvae were challenged with a high cholesterol diet (HCD; 4% w/w) for 10 days.\nLipoprotein injections showed impaired uptake of both LDL and oxLDL into the vessel wall of caudal veins of stab1-/-stab2-/- zebrafish, which was paralleled by redistribution to tissue macrophages. Total body cholesterol levels did not differ between HCD-fed stab1-/-stab2-/- and ABTL zebrafish. However, stab1-/-stab2-/- larvae exhibited 1.4-fold higher mRNA expression levels of ldlra involved in (modified) LDL uptake, whereas the expression levels of scavenger receptors scarb1 and cd36 were significantly decreased.\nWe have shown that stabilins 1 and 2 have an important scavenging function for apolipoprotein B-containing lipoproteins in zebrafish and that combined deficiency of these two proteins strongly upregulates the clearance of lipoproteins by macrophages within the caudal vein. Our current study highlights the use of zebrafish as model to study lipoprotein metabolism and liver sinusoidal endothelial cell function.Drug Delivery Technolog

    Application of zebrafish and murine models in lipoprotein metabolism and atherosclerosis research

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    Cardiovascular diseases are still a major concern for the global health. The main underlying pathology of this disease is atherosclerosis which is characterized by the accumulation of lipids and immune cells in the arterial wall leading to a chronic local inflammation and lesion formation. In this thesis, we aimed to (1) validate the use of zebrafish in cholesterol metabolism and atherosclerosis research, (2) study the role of certain classes of scavenger receptors in lipoprotein uptake and cholesterol-based functions, and (3) validated two immune-based potential targets for atherosclerosis.</p

    Vulnerable plaque and vulnerable blood: Two critical factors for spontaneous atherothrombosis in mouse models

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    Atherothrombotic events such as myocardial infarction and ischemic stroke are a major cause of morbidity and mortality worldwide. Understanding the molecular and cellular mechanisms of atherosclerotic plaque destabilization or erosion, and developing new therapeutics to prevent acute cardiovascular events is important for vascular biology research and clinical cardiovascular medicine. However, basic research on plaque destabilization, rupture and erosion is hampered by the lack of appropriate animal models of atherothrombosis. Unprovoked atherothrombosis is very scarce in commonly used mouse models for atherosclerosis, the low-density lipoprotein receptor knockout and apolipoprotein E knockout mice. Therefore, specific interventions are required to induce atherothrombosis in these models. Two strategies can be employed to induce atherothrombosis: 1) plaque destabilization and 2) induction of blood hypercoagulability. Although the individual strategies yield atherothrombosis at low incidence, it appears that the combination of both plaque destabilization and an increase in blood coagulability is the most promising strategy to induce atherothrombosis on a larger scale. In this review, we summarize the recent developments on mouse models for the investigation of atherothrombosis.Biopharmaceutic

    The role of endothelial and macrophage scavenger receptor stabilin 1 and 2 in early atherosclerosis development in zebrafish

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    Atherosclerosis, the main underlying cause of cardiovascular disease and related morbidities, is a multifactorial disease in which liver endothelial cells and macrophages have shown to play an important role. Recently, it has been demonstrated that oxidized phospholipids in oxidized LDL (oxLDL) are pro-inflammatory and pro-atherogenic in hypercholesterolaemic mice. Multiple scavenger receptors are known for oxLDL. They form a superfamily of membrane bound receptors that bind and internalize different types of ligands including other modified lipoproteins, endogenous proteins and pathogens. Several studies, including a microarray study of our research group, have already identified scavenger receptors stabilin-1 and -2 (STAB1 and STAB2) as a risk factor for atherosclerosis, and therefore as potential targets for treatment. An upcoming and powerful model organism to study early atherogenesis and that allows non-invasive cell tracking studies is the zebrafish. Recent studies have demonstrated the possibility to induce (early) atherosclerosis in zebrafish.We have used Crispr/Cas9 mutagenesis to generate zebrafish stab1 and stab2 mutants.Our results show that oxLDL clearance by zebrafish scavenger endothelial cells (homologous to liver sinusoidal endothelial cells) is diminished in stab2 zebrafish mutants and is absent in stab1/2 double mutants. In stabilin mutants, macrophage oxLDL uptake is increased, suggesting a protective role for stabilins in foam cell formation.To study this further, we are currently performing an atherosclerosis study to examine foam cell formation and vascular lipid accumulation in stabilin mutant zebrafish.Biopharmaceutic

    Silencing anticoagulant protein c causes spontaneous atherothrombosis in ApoE knockout mice

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    Objectives: Studies on atherothrombosis, a major cause of cardiovascularevents, are hampered by the lack of animal models spontaneously developingatherothrombosis. Major underlying problems are the stability ofthe atherosclerotic plaques, combined with the strong plasma anticoagulantactivity. We hypothesized that inhibition of the anticoagulant systemby silencing of protein C (Proc) may cause murine atherosclerotic plaquesto become prone to atherothrombosis.Biopharmaceutic

    Elimination of adrenocortical apolipoprotein E production does not impact glucocorticoid output in wild-type mice

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    Apolipoprotein E (APOE) deficient mice exhibit unexplained hypercorticosteronemia. Given that APOE is also produced locally within the adrenals, we evaluated the effect of adrenal-specific APOE deficiency on the glucocorticoid function. Hereto, one adrenal containing or lacking APOE was transplanted into adrenalectomized wild-type mice. Adrenal APOE deficiency did not impact adrenal total cholesterol levels. Importantly, the ability of the two adrenal types to produce glucocorticoids was also not different as judged from the similar plasma corticosterone levels. Adrenal mRNA expression levels of HMG-CoA reductase and the LDL receptor were decreased by respectively 72% (p < 0.01) and 65% (p = 0.07), suggesting that cholesterol acquisition pathways were inhibited to possibly compensate the lack of APOE. In support, a parallel increase in the expression level of the cholesterol accumulation-associated ER stress marker CHOP was detected (+117%; p < 0.05). In conclusion, our studies show that elimination of adrenocortical APOE production does not impact glucocorticoid output in wild-type mice.Biopharmaceutic
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