Posttranscriptional Regulation of the Human LDL Receptor by the U2-Spliceosome,Circulation Research

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Posttranscriptional Regulation of the Human LDL Receptor by the U2-Spliceosome
Circulation Research ( IF 16.5 ) Pub Date : 2021-11-23 , DOI: 10.1161/circresaha.120.318141
Paolo Zanoni _{1,

2,

3} , Grigorios Panteloglou _{1,

3} , Alaa Othman ₄ , Joel T Haas ₅ , Roger Meier ₆ , Antoine Rimbert _{7,

8} , Marta Futema ₉ , Yara Abou Khalil _{10,

11} , Simon F Norrelykke ₆ , Andrzej J Rzepiela ₆ , Szymon Stoma ₆ , Michael Stebler ₆ , Freerk van Dijk ₁₂ , Melinde Wijers ₇ , Justina C Wolters ₇ , Nawar Dalila ₁₃ , Nicolette C A Huijkman ₇ , Marieke Smit ₇ , Antonio Gallo ₁₄ , Valérie Carreau ₁₄ , Anne Philippi ₁₅ , Jean-Pierre Rabès _{10,

16,

17} , Catherine Boileau _{10,

18} , Michele Visentin ₁₉ , Luisa Vonghia _{20,

21} , Jonas Weyler ₂₀ , Sven Francque _{20,

21} , An Verrijken _{21,

22} , Ann Verhaegen _{21,

22} , Luc Van Gaal _{21,

22} , Adriaan van der Graaf ₁₂ , Belle V van Rosmalen ₂₃ , Jerome Robert ₁ , Srividya Velagapudi _{1,

3,

24} , Mustafa Yalcinkaya _{1,

3,

25} , Michaela Keel _{1,

3} , Silvija Radosavljevic _{1,

3} , Andreas Geier ₂₆ , Anne Tybjaerg-Hansen ₁₃ , Mathilde Varret ₁₀ , Lucia Rohrer _{1,

3} , Steve E Humphries ₂₇ , Bart Staels ₅ , Bart van de Sluis ₇ , Jan Albert Kuivenhoven ₇ , Arnold von Eckardstein _{1,

3}

Affiliation

Institute for Clinical Chemistry, University and University Hospital Zurich, Switzerland (P.Z., G.P., J.R., S.V., M.Y., M.K., S.R., L.R., A.v.E.).
Now with Institute of Medical Genetics, University of Zurich, Switzerland (P.Z.).
Center for Integrative Human Physiology, University of Zurich, Switzerland (P.Z., G.P., S.V., M.Y., M.K., S.R., L.R., A.v.E.).
Institute of Molecular Systems Biology, ETH Zurich, Switzerland (A.O.).
University of Lille, Inserm, CHU Lille, Institut Pasteur de Lille, U1011- EGID, France (J.T.H., B.S.).
Scientific center for optical and electron microscopy (ScopeM), ETH Zurich, Switzerland (R.M., S.F.N., A.J.R., S.S., M. Stebler).
Department of Pediatrics, Section Molecular Genetics, University of Groningen, University Medical Center Groningen, the Netherlands (A.R., M.W., J.C.W., N.C.A.H., M. Smit, B.v.d.S., J.A.K.).
Now with Inserm UMR 1087/CNRS UMR 6291 IRS-UN, Nantes, France (A.R.).
Cardiology Research Centre, Molecular and Clinical Sciences Research Institute, St George's, University of London, United Kingdom (M.F.).
LVTS-INSERM UMRS 1148 and University of Paris, CHU Xavier Bichat, Paris, France (Y.A.K., J.-P.R., C.B., M. Varret).
Laboratory of Biochemistry and Molecular Therapeutics (LBTM), Faculty of Pharmacy and Pôle technologie Santé (PTS), Saint-Joseph University, Beirut, Lebanon (Y.A.K.).
Department of Genetics, University of Groningen, University Medical Center Groningen, the Netherlands (F.v.D., A.v.d.G.).
Department of Clinical Biochemistry, Rigshospitalet, Copenhagen University Hospital, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark (N.D., A.T.-H.).
AP-HP, Endocrinology and Metabolism Department, Human Research Nutrition Center, Pitié-Salpêtrière Hospital, Paris, France (A. Gallo, V.C.).
Université de Paris, Faculté de Médecine Paris-Diderot, UMR-S958 Paris, France; Now with Université de Paris, Institut Cochin, INSERM U1016, CNRS UMR-8104, Paris, France (A.P.).
AP-HP, Université Paris-Saclay, Paris, France (J.-P.R.).
UFR Simone Veil des Sciences de la Santé, UVSQ, Montigny-Le-Bretonneux, France (J.-P.R.).
AP-HP, Genetics Department, CHU Xavier Bichat, Université de Paris, France (C.B.).
Department of Clinical Pharmacology and Toxicology, University Hospital Zurich, Switzerland (M. Visentin).
Department of Gastroenterology and Hepatology, Antwerp University Hospital, Edegem, Belgium (L.V., J.W., S.F.).
Laboratory of Experimental Medicine and Paediatrics, Faculty of Medicine, University of Antwerp, Belgium (L.V., J.W., S.F., A. Verrijken, A. Verhaegen, L.V.G.).
Department of Endocrinology, Diabetology and Metabolism, Antwerp University Hospital, Edegem, Belgium (A. Verrijken, A. Verhaegen, L.V.G.).
Department of Surgery, Academic Medical Center, University of Amsterdam, the Netherlands (B.V.v.R.).
Center for Molecular Cardiology, University of Zurich, Switzerland (S.V.).
Division of Molecular Medicine, Department of Medicine, Columbia University, New York, NY (M.Y.).
Division of Hepatology, Department of Medicine II, University Hospital Würzburg, Germany (A. Geier).
Cardiovascular Genetics, Institute of Cardiovascular Science, University College London, United Kingdom (S.E.H.).

Background:The LDLR (low-density lipoprotein receptor) in the liver is the major determinant of LDL-cholesterol levels in human plasma. The discovery of genes that regulate the activity of LDLR helps to identify pathomechanisms of hypercholesterolemia and novel therapeutic targets against atherosclerotic cardiovascular disease.Methods:We performed a genome-wide RNA interference screen for genes limiting the uptake of fluorescent LDL into Huh-7 hepatocarcinoma cells. Top hit genes were validated by in vitro experiments as well as analyses of data sets on gene expression and variants in human populations.Results:The knockdown of 54 genes significantly inhibited LDL uptake. Fifteen of them encode for components or interactors of the U2-spliceosome. Knocking down any one of 11 out of 15 genes resulted in the selective retention of intron 3 of LDLR. The translated LDLR fragment lacks 88% of the full length LDLR and is detectable neither in nontransfected cells nor in human plasma. The hepatic expression of the intron 3 retention transcript is increased in nonalcoholic fatty liver disease as well as after bariatric surgery. Its expression in blood cells correlates with LDL-cholesterol and age. Single nucleotide polymorphisms and 3 rare variants of one spliceosome gene, RBM25, are associated with LDL-cholesterol in the population and familial hypercholesterolemia, respectively. Compared with overexpression of wild-type RBM25, overexpression of the 3 rare RBM25 mutants in Huh-7 cells led to lower LDL uptake.Conclusions:We identified a novel mechanism of posttranscriptional regulation of LDLR activity in humans and associations of genetic variants of RBM25 with LDL-cholesterol levels.

中文翻译：

U2剪接体对人低密度脂蛋白受体的转录后调控

背景：肝脏中的 LDLR（低密度脂蛋白受体）是人血浆中 LDL-胆固醇水平的主要决定因素。调节 LDLR 活性的基因的发现有助于确定高胆固醇血症的病理机制和针对动脉粥样硬化性心血管疾病的新治疗靶点。方法：我们对限制荧光 LDL 摄取到 Huh-7 肝癌细胞的基因进行了全基因组 RNA 干扰筛选. 热门基因通过体外实验以及人群中基因表达和变异的数据集分析得到验证。结果：54 个基因的敲低显着抑制了 LDL 摄取。其中 15 个编码 U2 剪接体的成分或相互作用物。敲除 15 个基因中的 11 个基因中的任何一个，都会导致内含子 3 的选择性保留。低密度脂蛋白。翻译后的 LDLR 片段缺少全长 LDLR 的 88%，在未转染的细胞和人血浆中都无法检测到。内含子 3 保留转录物的肝脏表达在非酒精性脂肪性肝病以及减肥手术后增加。它在血细胞中的表达与低密度脂蛋白胆固醇和年龄相关。一个剪接体基因RBM25的单核苷酸多态性和 3 个罕见变体分别与人群中的低密度脂蛋白胆固醇和家族性高胆固醇血症有关。与野生型RBM25的过表达相比，3 种罕见的RBM25过表达Huh-7 细胞中的突变体导致 LDL 摄取降低。结论：我们确定了人类 LDLR 活性转录后调控的新机制以及RBM25遗传变异与 LDL-胆固醇水平的关联。

更新日期：2022-01-08

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