Posttranscriptional Regulation of the Human LDL Receptor by the U2-Spliceosome

Paolo Zanoni^#^{1

2

3}, Grigorios Panteloglou^#^{1

3}, Alaa Othman⁴, Joel T Haas⁵, Roger Meier⁶, Antoine Rimbert^{7

8}, Marta Futema⁹, Yara Abou Khalil^{10

11}, Simon F Norrelykke⁶, Andrzej J Rzepiela⁶, Szymon Stoma⁶, Michael Stebler⁶, Freerk van Dijk¹², Melinde Wijers⁷, Justina C Wolters⁷, Nawar Dalila¹³, Nicolette C A Huijkman⁷, Marieke Smit⁷, Antonio Gallo¹⁴, Valérie Carreau¹⁴, Anne Philippi¹⁵, Jean-Pierre Rabès^{10

16

17}, Catherine Boileau^{10

18}, Michele Visentin¹⁹, Luisa Vonghia^{20

21}, Jonas Weyler²⁰, Sven Francque^{20

21}, An Verrijken^{21

22}, Ann Verhaegen^{21

22}, Luc Van Gaal^{21

22}, Adriaan van der Graaf¹², Belle V van Rosmalen²³, Jerome Robert¹, Srividya Velagapudi^{1

24

3}, Mustafa Yalcinkaya^{1

25

3}, Michaela Keel^{1

3}, Silvija Radosavljevic^{1

3}, Andreas Geier²⁶, Anne Tybjaerg-Hansen¹³, Mathilde Varret¹⁰, Lucia Rohrer^{1

3}, Steve E Humphries²⁷, Bart Staels⁵, Bart van de Sluis⁷, Jan Albert Kuivenhoven⁷, Arnold von Eckardstein^{1

3}

Affiliations

¹ 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.).
² Now with Institute of Medical Genetics, University of Zurich, Switzerland (P.Z.).
³ 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.).
⁴ Institute of Molecular Systems Biology, ETH Zurich, Switzerland (A.O.).
⁵ University of Lille, Inserm, CHU Lille, Institut Pasteur de Lille, U1011- EGID, France (J.T.H., B.S.).
⁶ Scientific center for optical and electron microscopy (ScopeM), ETH Zurich, Switzerland (R.M., S.F.N., A.J.R., S.S., M. Stebler).
⁷ 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.).
⁸ Now with Inserm UMR 1087/CNRS UMR 6291 IRS-UN, Nantes, France (A.R.).
⁹ Cardiology Research Centre, Molecular and Clinical Sciences Research Institute, St George's, University of London, United Kingdom (M.F.).
¹⁰ LVTS-INSERM UMRS 1148 and University of Paris, CHU Xavier Bichat, Paris, France (Y.A.K., J.-P.R., C.B., M. Varret).
¹¹ Laboratory of Biochemistry and Molecular Therapeutics (LBTM), Faculty of Pharmacy and Pôle technologie Santé (PTS), Saint-Joseph University, Beirut, Lebanon (Y.A.K.).
¹² Department of Genetics, University of Groningen, University Medical Center Groningen, the Netherlands (F.v.D., A.v.d.G.).
¹³ Department of Clinical Biochemistry, Rigshospitalet, Copenhagen University Hospital, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark (N.D., A.T.-H.).
¹⁴ AP-HP, Endocrinology and Metabolism Department, Human Research Nutrition Center, Pitié-Salpêtrière Hospital, Paris, France (A. Gallo, V.C.).
¹⁵ 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.).
¹⁶ AP-HP, Université Paris-Saclay, Paris, France (J.-P.R.).
¹⁷ UFR Simone Veil des Sciences de la Santé, UVSQ, Montigny-Le-Bretonneux, France (J.-P.R.).
¹⁸ AP-HP, Genetics Department, CHU Xavier Bichat, Université de Paris, France (C.B.).
¹⁹ Department of Clinical Pharmacology and Toxicology, University Hospital Zurich, Switzerland (M. Visentin).
²⁰ Department of Gastroenterology and Hepatology, Antwerp University Hospital, Edegem, Belgium (L.V., J.W., S.F.).
²¹ 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.).
²² Department of Endocrinology, Diabetology and Metabolism, Antwerp University Hospital, Edegem, Belgium (A. Verrijken, A. Verhaegen, L.V.G.).
²³ Department of Surgery, Academic Medical Center, University of Amsterdam, the Netherlands (B.V.v.R.).
²⁴ Center for Molecular Cardiology, University of Zurich, Switzerland (S.V.).
²⁵ Division of Molecular Medicine, Department of Medicine, Columbia University, New York, NY (M.Y.).
²⁶ Division of Hepatology, Department of Medicine II, University Hospital Würzburg, Germany (A. Geier).
²⁷ Cardiovascular Genetics, Institute of Cardiovascular Science, University College London, United Kingdom (S.E.H.).

^# Contributed equally.

PMID: 34809444
DOI: 10.1161/CIRCRESAHA.120.318141

Free article

Posttranscriptional Regulation of the Human LDL Receptor by the U2-Spliceosome

Paolo Zanoni et al. Circ Res. 2022.

Free article

. 2022 Jan 7;130(1):80-95.

doi: 10.1161/CIRCRESAHA.120.318141. Epub 2021 Nov 23.

Authors

Affiliations

¹ 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.).
² Now with Institute of Medical Genetics, University of Zurich, Switzerland (P.Z.).
³ 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.).
⁴ Institute of Molecular Systems Biology, ETH Zurich, Switzerland (A.O.).
⁵ University of Lille, Inserm, CHU Lille, Institut Pasteur de Lille, U1011- EGID, France (J.T.H., B.S.).
⁶ Scientific center for optical and electron microscopy (ScopeM), ETH Zurich, Switzerland (R.M., S.F.N., A.J.R., S.S., M. Stebler).
⁷ 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.).
⁸ Now with Inserm UMR 1087/CNRS UMR 6291 IRS-UN, Nantes, France (A.R.).
⁹ Cardiology Research Centre, Molecular and Clinical Sciences Research Institute, St George's, University of London, United Kingdom (M.F.).
¹⁰ LVTS-INSERM UMRS 1148 and University of Paris, CHU Xavier Bichat, Paris, France (Y.A.K., J.-P.R., C.B., M. Varret).
¹¹ Laboratory of Biochemistry and Molecular Therapeutics (LBTM), Faculty of Pharmacy and Pôle technologie Santé (PTS), Saint-Joseph University, Beirut, Lebanon (Y.A.K.).
¹² Department of Genetics, University of Groningen, University Medical Center Groningen, the Netherlands (F.v.D., A.v.d.G.).
¹³ Department of Clinical Biochemistry, Rigshospitalet, Copenhagen University Hospital, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark (N.D., A.T.-H.).
¹⁴ AP-HP, Endocrinology and Metabolism Department, Human Research Nutrition Center, Pitié-Salpêtrière Hospital, Paris, France (A. Gallo, V.C.).
¹⁵ 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.).
¹⁶ AP-HP, Université Paris-Saclay, Paris, France (J.-P.R.).
¹⁷ UFR Simone Veil des Sciences de la Santé, UVSQ, Montigny-Le-Bretonneux, France (J.-P.R.).
¹⁸ AP-HP, Genetics Department, CHU Xavier Bichat, Université de Paris, France (C.B.).
¹⁹ Department of Clinical Pharmacology and Toxicology, University Hospital Zurich, Switzerland (M. Visentin).
²⁰ Department of Gastroenterology and Hepatology, Antwerp University Hospital, Edegem, Belgium (L.V., J.W., S.F.).
²¹ 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.).
²² Department of Endocrinology, Diabetology and Metabolism, Antwerp University Hospital, Edegem, Belgium (A. Verrijken, A. Verhaegen, L.V.G.).
²³ Department of Surgery, Academic Medical Center, University of Amsterdam, the Netherlands (B.V.v.R.).
²⁴ Center for Molecular Cardiology, University of Zurich, Switzerland (S.V.).
²⁵ Division of Molecular Medicine, Department of Medicine, Columbia University, New York, NY (M.Y.).
²⁶ Division of Hepatology, Department of Medicine II, University Hospital Würzburg, Germany (A. Geier).
²⁷ Cardiovascular Genetics, Institute of Cardiovascular Science, University College London, United Kingdom (S.E.H.).

^# Contributed equally.

PMID: 34809444
DOI: 10.1161/CIRCRESAHA.120.318141

Abstract

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.

Keywords: cardiovascular diseases; endocytosis; hepatocytes; hypercholesterolemia; spliceosomes.

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Posttranscriptional Regulation of the Human LDL Receptor by the U2-Spliceosome

Affiliations

Posttranscriptional Regulation of the Human LDL Receptor by the U2-Spliceosome

Authors

Affiliations

Abstract

Publication types

MeSH terms

Substances

Grants and funding

LinkOut - more resources

Full Text Sources

Molecular Biology Databases