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Cited by 27

The DOI matches its Crossref record; 5 of the 5 references it cites were found in a registry or an index.

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

Zanoni, Paolo, Panteloglou, Grigorios, Othman, Alaa, Haas, Joel T., Meier, Roger, Rimbert, Antoine, Futema, Marta, Abou Khalil, Yara, Norrelykke, Simon F., Rzepiela, Andrzej J., Stoma, Szymon, Stebler, Michael, van Dijk, Freerk, Wijers, Melinde, Wolters, Justina C., Dalila, Nawar, Huijkman, Nicolette C. A., Smit, Marieke, Gallo, Antonio, Carreau, Valérie, Philippi, Anne, Rabès, Jean-Pierre, Boileau, Catherine, Visentin, Michele, Vonghia, Luisa, Weyler, Jonas, Francque, Sven, Verrijken, An, Verhaegen, Ann, Van Gaal, Luc, van der Graaf, Adriaan, van Rosmalen, Belle V., Robert, Jerome, Velagapudi, Srividya, Yalcinkaya, Mustafa, Keel, Michaela, Radosavljevic, Silvija, Geier, Andreas, Tybjaerg-Hansen, Anne, Varret, Mathilde, Rohrer, Lucia, Humphries, Steve E., Staels, Bart, van de Sluis, Bart, Kuivenhoven, Jan Albert, von Eckardstein, Arnold·2022·Circulation Research
DOI 10.1161/circresaha.120.318141
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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.

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Crossref · 10.1161/circresaha.120.318141

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  • LDLR mRNA is regulated via multiple microRNAs, alternative splicing to produce non-functional LDLR protein, and regulation of LDLR mRNA decay.

    “At the post-transcriptional level, LDLR mRNA is regulated via several mechanisms including multiple microRNAs [30], alternative splicing to produce non-functional LDLR protein [31], and regulation of LDLR mRNA decay [32].”

    From Post-translational regulation of the low-density lipoprotein receptor provides new targets for cholesterol regulation (2024) · cites this paper as [31]

    The Discussion (page unavailable) reports that intron 3 retention produces a truncated LDLR receptor, supporting the alternative-splicing mechanism attributed to citation [31]. It describes that receptor as “most probably non-functional,” whereas the citing sentence states non-functionality without qualification. The microRNA and mRNA-decay clauses have separate citations and are not judged here. (LLM, conf=0.83)

    Experimental — accuracy not yet measured

  • Experimental evidence has recapitulated key modulators of cellular LDL uptake, as well as posttranscriptional and posttranslational regulators of LDLR.

    “Moreover, experimental evidence has recapitulated key modulators of cellular LDL uptake, as well as posttranscriptional and posttranslational regulators of LDLR79–82.”

    From ABCC4 impairs the clearance of plasma LDL cholesterol through suppressing LDLR expression in the liver (2025) · cites this paper as [82]

    The Results report that the RNAi screen identified genes affecting cellular LDL uptake, and the Discussion describes experimental validation of U2-spliceosome components as posttranscriptional regulators of LDLR. But the Discussion (p. 9) says the screen missed canonical regulators including MYLIP and PCSK9, so this paper does not support the claim that its experiments recapitulated posttranslational LDLR regulators. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • A whole genome RNA-interference screen in Huh7 cells revealed the involvement of 15 U2-spliceosome components in LDL uptake.

    “A whole genome RNA-interference screen in Huh7, a human hepatic carcinoma cell line, aimed at studying LDL uptake revealed the involvement of 15 U2-spliceosome components (Fig. 2) [18].”

    From The low-density lipoprotein receptor: Emerging post-transcriptional regulatory mechanisms (2025) · cites this paper as [18]

    The Introduction reports a genome-wide RNAi screen in Huh-7 cells and identifies 15 genes whose knockdown reduced LDL uptake. The Results distinguish six core U2-spliceosome components from other proteins that interact with the U2-spliceosome, so the source does not establish that all 15 are components. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • Silencing the expression of 11 U2-spliceosome genes significantly increases retention of intron 3 in LDLR mRNA without altering expression of the full-length transcript.

    “Silencing the expression of 11 of these genes significantly increase the retention of intron 3 in LDLR mRNA, without altering the expression of the full-length transcript.”

    From The low-density lipoprotein receptor: Emerging post-transcriptional regulatory mechanisms (2025) · cites this paper as [18]

    In “Loss of U2-spliceosome genes and their interactors causes selective retention of LDLR intron 3” (page unavailable), the RNA-sequencing results report increased intron 3 retention after knockdown of ten of the eleven genes, without altering full-length LDLR transcript expression. RBM25 was the exception in that analysis; a separate qRT-PCR assay found an increase after RBM25 knockdown, but the passage does not establish the claimed significant increase for all eleven genes. (LLM, conf=0.90)

    Experimental — accuracy not yet measured

  • RBM25 variants result in increased retention of intron 3 and are associated with elevated plasma LDL-cholesterol levels and potentially linked to unexplained familial hypercholesterolemia.

    “Additionally, this study identified variants in RBM25, a U2-spliceosome component that resulted in increased retention of intron 3. Consequently, these variants were associated with elevated plasma LDL-cholesterol levels and were potentially linked to cases of unexplained familial hypercholesterolemia (FH).”

    From The low-density lipoprotein receptor: Emerging post-transcriptional regulatory mechanisms (2025) · cites this paper as [18]

    The section on RBM25 variants (page not specified) reports that rare variants were enriched in patients with familial hypercholesterolemia unexplained by known FH genes. But it found no significant increase in LDLR intron 3 retention when those mutants were expressed, and the Discussion says their effect on LDL uptake may be unrelated to intron 3 retention. The paper also reports that the RBM25 SNPs associated with LDL-cholesterol in the UK Biobank were associated with lower, not elevated, levels. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

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Citations

Cites

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Metadata

Authors
Zanoni, P., Panteloglou, G., Othman, A., Haas, J. T., Meier, R., Rimbert, A., Futema, M., Abou Khalil, Y., Norrelykke, S. F., Rzepiela, A. J., Stoma, S., Stebler, M., van Dijk, F., Wijers, M., Wolters, J. C., Dalila, N., Huijkman, N. C. A., Smit, M., Gallo, A., Carreau, V., Philippi, A., Rabès, J.-P., Boileau, C., Visentin, M., Vonghia, L., Weyler, J., Francque, S., Verrijken, A., Verhaegen, A., Van Gaal, L., van der Graaf, A., van Rosmalen, B. V., Robert, J., Velagapudi, S., Yalcinkaya, M., Keel, M., Radosavljevic, S., Geier, A., Tybjaerg-Hansen, A., Varret, M., Rohrer, L., Humphries, S. E., Staels, B., van de Sluis, B., Kuivenhoven, J. A., & von Eckardstein, A.
Affiliations
  • 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.)
  • 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.)
  • 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).
  • 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.).
  • 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 Surgery, Academic Medical Center, University of Amsterdam, the Netherlands (B.V.v.R.).
  • 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.).
ORCID
Venue
Circulation Research
Publisher
Ovid Technologies (Wolters Kluwer Health)
Published
2022-01-07
Volume
130
Issue
1
Pages
80-95
ISSN
  • 0009-7330
  • 1524-4571
DOI
10.1161/circresaha.120.318141
Cited by
27
References
5
Language
en
Funders
  • EC | FP7 | FP7 Health (603091, 305707)
  • European Genomic Institute for Diabetes (ANR-10-LABX-46)
  • Swiss National Science Foundation (31003A-160126, 310030-185109)
  • Dutch Heart Foundation (2015T068)
  • Swiss Atherosclerosis Society (AGLA Award)
  • DACH Gesellschaft für Praevention (DACH Award)
  • GeniusII (CVON2017-2020)
  • Fondation Maladies Rares (AOM06024)
  • Agence Nationale de la Recherche (ANR-16-RHUS-0007, ANR 16-RHUS-0006 (PreciNASH ))
  • EMBO (ALTF277-2014)
  • EC | European Research Council (694717)
  • EC | Sixth Framework Programme (LSHM-CT-2005-018734)
  • Fonds Wetenschappelijk Onderzoek (1802154N)
  • Ministere de l'Education Nationale et de la Technologie (NA)
  • British Heart Foundation (RG3008, PG008/08)
  • NIHR | UCLH Biomedical Research Centre (NN)
  • Forschungskommission University of Zurich (FK-20-037)
  • International Atherosclerosis Society (24052829)

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