Apolipoprotein E Deficiency Increases Remnant Lipoproteins and Accelerates Progressive Atherosclerosis, But Not Xanthoma Formation, in Gene-Modified Minipigs

Shim, Jeong; Poulsen, Christian Bo; Hagensen, Mette K.; Larsen, Torben; Heegaard, Peter M. H.; Christoffersen, Christina; Bolund, Lars; Schmidt, M.; Liu, Ying; Li, Juan; Li, Rong; Callesen, Henrik; Bentzon, Jacob F.; Sørensen, Charlotte Brandt

doi:10.1016/j.jacbts.2017.06.004

Cited by 10 publications

(6 citation statements)

References 28 publications

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“…It is interesting to note that food-derived remnant cholesterol has a modest contribution to the total cholesterol level. Moreover, it was previously reported that remnant lipoproteinemia in APOE Ϫ/Ϫ minipigs is not efficient in initiating atherosclerosis but contributes to the atherosclerotic progression of preexisting lesions (28). Therefore, the miR-27a mimic may diminish overall plasma total cholesterol levels in high-cholesterol-diet-fed Apoe Ϫ/Ϫ mice by modulating LDL, HDL, and other cholesterol fractions.…”

Section: Discussionmentioning

confidence: 99%

MicroRNA 27a Is a Key Modulator of Cholesterol Biosynthesis

Khan

Agarwal

Reddy

et al. 2020

Molecular and Cellular Biology

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Hypercholesterolemia is a strong predictor of cardiovascular diseases. The 3-hydroxy-3-methylglutaryl coenzyme A reductase gene (Hmgcr) coding for the rate-limiting enzyme in the cholesterol biosynthesis pathway is a crucial regulator of plasma cholesterol levels. However, the posttranscriptional regulation of Hmgcr remains poorly understood. The main objective of this study was to explore the role of microRNAs (miRNAs) in the regulation of Hmgcr expression. Systematic in silico predictions and experimental analyses reveal that miRNA 27a (miR-27a) specifically interacts with the Hmgcr 3′ untranslated region in murine and human hepatocytes. Moreover, our data show that Hmgcr expression is inversely correlated with miR-27a levels in various cultured cell lines and in human and rodent tissues. Actinomycin D chase assays and relevant experiments demonstrate that miR-27a regulates Hmgcr by translational attenuation followed by mRNA degradation. Early growth response 1 (Egr1) regulates miR-27a expression under basal and cholesterol-modulated conditions. miR-27a augmentation via tail vein injection of miR-27a mimic in high-cholesterol-diet-fed Apoe−/− mice shows downregulation of hepatic Hmgcr and plasma cholesterol levels. Pathway and gene expression analyses show that miR-27a also targets several other genes (apart from Hmgcr) in the cholesterol biosynthesis pathway. Taken together, miR-27a emerges as a key regulator of cholesterol biosynthesis and has therapeutic potential for the clinical management of hypercholesterolemia.

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Section: Discussionmentioning

confidence: 99%

MicroRNA 27a Is a Key Modulator of Cholesterol Biosynthesis

Khan

Agarwal

Reddy

et al. 2020

Molecular and Cellular Biology

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“…The genome of the μMP has also been partially sequenced and the genomic information further facilitates atherosclerosis studies (31). In addition, knockout swine using domestic and miniature pigs are currently available for atherosclerosis research, including LDLr and/or apolipoprotein E knockout swine reproducing human atherogenesis (32)(33)(34)(35)(36). The body size of these domestic and traditional miniature pigs, however, is still too big to be maintained and handled in an ordinary laboratory, especially for long-term experiments.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, μMPs compensate for the disadvantages of body size; cloned-μMPs, as well as noncloned-μMP, would be a valuable tool for human atherosclerosis research and related diseases. As in the cases with other swine strains (32)(33)(34)(35)(36), transgenic and knockout μMPs may be produced by gene modification techniques in somatic cell nuclear transfer or fertilized eggs. The development of swine embryonal stem cells would be another advancement for the production of gene-modified-μMPs, possibly as a more efficient method than the somatic cell nuclear transfer.…”

Section: Discussionmentioning

confidence: 99%

High Pathological Reproducibility of Diet-induced Atherosclerosis in MicrominipigsviaCloning Technology

Otake

Kawaguchi

Enya

et al. 2021

In Vivo

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Background/Aim: The reproducibility of atherosclerotic lesions was evaluated after the production of clonedmicrominipigs and their offspring. Materials and Methods: Cloned-microminipig-parents were produced by microminipigsomatic cell nuclei. These parents were crossbred and delivered males (F1-offspring) were divided into two groups: normal chow diet (NcD)-fed and high-fat/high-cholesterol diet (HcD)-fed groups. One of the F1-offsprings was subjected to cloning, and delivered males (F1-clones) were fed with HcD. After 8 weeks, all animals were necropsied for pathophysiological studies compared to non-cloned-microminipigs. Results: HcD-fed F1-offspring and F1-clones, but not NcDfed F1-offspring, exhibited increased serum lipid levels and systemic atherosclerosis, which were comparable to those of HcD-fed non-cloned-microminipigs. Homogeneity of variance analysis demonstrated that standard deviation values of serum lipoprotein and aortic atherosclerosis area from HcD-fed animals decreased in F1-offspring and F1-clones. Conclusion: HcD-induced atherogenesis was highly reproducible in F1offsprings and F1-clones, indicating that the atherosclerosisprone genomic background was preserved in the clonedmicrominipigs, which can be used for studies on human atherosclerosis and related diseases.To study the pathogenesis of atherosclerosis, many animals, including mice, rats, rabbits, and swine are widely used worldwide (1). In contrast to human lipid metabolism, mice and rats are high-density lipoprotein (HDL)-cholesterol dominant in their serum lipid profiles and are essentially resistant to the high-fat/high-cholesterol diet (HcD) used to develop atherosclerosis. This is partially explained by the absence of expression of several genes that regulate lipid metabolism and atherogenesis (1-3). Therefore, gene modification, for example gene knockout of apolipoprotein E or low-density lipoprotein receptor (LDLr), is necessary to efficiently produce atherosclerosis in mice (1, 4). However, since the pathogenesis of atherosclerosis includes both genetic and environmental factors, animal models should ideally have anatomy, physiology, and life habits similar to humans (5). In this sense, swine are a potentially useful animal model for atherosclerosis research because their anatomy, physiology, lipid metabolism, and life habits are very similar to those of humans (6, 7). 2025This article is freely accessible online.

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“…Genetic studies have revealed that variants in ApoE are associated with the susceptibility to coronary heart disease and stroke (Bennet et al, 2007;Khan et al, 2013). ApoE knockout (KO) mice, rats, rabbits, and pigs show increased levels of cholesterols together with apparent atherosclerosis in large arteries such as the aorta (Zhang et al, 1992;Plump et al, 1992;Shim et al, 2017;Fang et al, 2018;Niimi et al, 2016;Zhao et al, 2018). However, clinical manifestations associated with advanced atherosclerosis have been rarely reported in these ApoE mutants.…”

Section: Introductionmentioning

confidence: 99%

Dogs lacking Apolipoprotein E show advanced atherosclerosis leading to apparent clinical complications

Zhao

et al. 2021

Sci. China Life Sci.

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Atherosclerotic cardiovascular disease resulting from dysregulated lipid metabolism is the leading cause of morbidity and mortality worldwide. Apolipoprotein E (ApoE) plays a critical role in cholesterol metabolism. Knockouts in lipid-metabolizing proteins including ApoE in multiple model organisms such as mice and rats exhibiting elevated levels of cholesterol have been widely used for dissecting the pathology of atherosclerosis, but few of these animal models exhibit advanced atherosclerotic plaques leading to ischemia-induced clinical symptoms, limiting their use for translational studies. Here we report hypercholesterolemia and severe atherosclerosis characterized by stenosis and occlusion of arteries, together with clinical manifestations of stroke and gangrene, in ApoE knockout dogs generated by CRISPR/Cas9 and cloned by somatic cell nuclear transfer technologies. Importantly, the hypercholesterolemia and atherosclerotic complications in F0 mutants are recapitulated in their offspring. As the ApoE-associated atherosclerosis and clinical manifestations in mutant dogs are more similar to that in human patients compared with those in other animal models, these mutant dogs will be invaluable in developing and evaluating new therapies, including endovascular procedures, against atherosclerosis and related disorders.

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Apolipoprotein E Deficiency Increases Remnant Lipoproteins and Accelerates Progressive Atherosclerosis, But Not Xanthoma Formation, in Gene-Modified Minipigs

Abstract: Visual Abstract

Cited by 10 publications

References 28 publications

MicroRNA 27a Is a Key Modulator of Cholesterol Biosynthesis

MicroRNA 27a Is a Key Modulator of Cholesterol Biosynthesis

High Pathological Reproducibility of Diet-induced Atherosclerosis in MicrominipigsviaCloning Technology

Dogs lacking Apolipoprotein E show advanced atherosclerosis leading to apparent clinical complications

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