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

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

doi:10.1161/CIRCRESAHA.120.318141

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

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

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

12 Citationer (Scopus)

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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.

Originalsprog	Engelsk
Tidsskrift	Circulation Research
Vol/bind	130
Udgave nummer	1
Sider (fra-til)	80-95
Antal sider	16
ISSN	0009-7330
DOI	https://doi.org/10.1161/CIRCRESAHA.120.318141
Status	Udgivet - 2022

Bibliografisk note

Funding Information:
We acknowledge the use of data from BIOS-consortium ( http://wiki.bbmri.nl/wiki/BIOS_bios ) which is funded by BBMRI-NL (NWO project 184.021.007). Flow cytometry was performed with equipment of the flow cytometry facility, University of Zurich.

Funding Information:
This work was conducted as part of the TransCard project of the seventh Framework Program (FP7) granted by the European Commission, to J. Albert Kuivenhoven, A. Tybjaerg-Hansen, and A. von Eckardstein (number 603091) as well as partially the FP7 RESOLVE project (to J.T. Haas, B. Staels, A. Verhaegen, S. Francque, L. Van Gaal, and A. von Eckardstein) and the European Genomic Institute for Diabetes (EGID, ANR-10-LABX-46 to B. Staels). Additional work by A. von Eckardstein’s team was funded by the Swiss National Science Foundation (31003A-160126, 310030-185109) and the Swiss Systems X program (2014/267 [Medical Research and Development (MRD)] HDL-X). P. Zanoni received funding awards from the Swiss Atherosclerosis Society (Arbeitsgruppe Lipide und Atherosklerose [AGLA] and the DACH Society for Prevention of Cardiovascular Diseases). G. Panteloglou received funding from the University of Zurich (Forschungskredit, grant no. FK-20-037). J. Albert Kuivenhoven is an Established Investigator from the Dutch Heart Foundation (2015T068). J. Albert Kuivenhoven was also supported by GeniusII (CVON2017-2020). The Laboratory for Vascular Translational Science (L.V.T.S.) team is supported by Fondation Maladies Rares, Programme Hospitalier de Recherche Clinique (PHRC) (AOM06024), and the national project CHOPIN (CHolesterol Personalized Innovation), granted by the Agence Nationale de la Recherche (ANR-16-RHUS-0007). Y. Abou Khalil is supported by a grant from Ministère de l’Education Nationale et de la Technologie (France). J.T. Haas was supported by an EMBO Long Term Fellowship (ALTF277-2014). B. Staels is a recipient of an ERC Advanced Grant (no. 694717). Both are also supported by PreciNASH (ANR 16-RHUS-0006). Research at the Antwerp University Hospital was supported by the European Union: FP6 (HEPADIP Contract LSHM-CT-2005-018734). S. Francque has a senior clinical research fellowship from the Fund for Scientific Research (FWO) Flanders (1802154 N). S.E. Humphries received grants RG3008 and PG008/08 from the British Heart Foundation, and the support of the UCLH NIHR BRC. S.E. Humphries directs the UK Children’s FH Register which has been supported by a grant from Pfizer (24052829) given by the International Atherosclerosis Society.

Funding Information:
This work was conducted as part of the TransCard project of the seventh Framework Program (FP7) granted by the European Commission, to J. Albert Kuivenhoven, A. Tybjaerg-Hansen, and A. von Eckardstein (number 603091) as well as partially the FP7 RESOLVE project (to J.T. Haas, B. Staels, A. Verhaegen, S. Francque, L. Van Gaal, and A. von Eckardstein) and the European Genomic Institute for Diabetes (EGID, ANR-10-LABX-46 to B. Staels). Additional work by A. von Eckardstein's team was funded by the Swiss National Science Foundation (31003A-160126, 310030-185109) and the Swiss Systems X program (2014/267 [Medical Research and Development (MRD)] HDL-X). P. Zanoni received funding awards from the Swiss Atherosclerosis Society (Arbeitsgruppe Lipide und Atherosklerose [AGLA] and the DACH Society for Prevention of Cardiovascular Diseases). G. Panteloglou received funding from the University of Zurich (Forschungskredit, grant no. FK-20-037). J. Albert Kuivenhoven is an Established Investigator from the Dutch Heart Foundation (2015T068). J. Albert Kuivenhoven was also supported by GeniusII (CVON2017-2020). The Laboratory for Vascular Translational Science (L.V.T.S.) team is supported by Fondation Maladies Rares, Programme Hospitalier de Recherche Clinique (PHRC) (AOM06024), and the national project CHOPIN (CHolesterol Personalized Innovation), granted by the Agence Nationale de la Recherche (ANR-16-RHUS-0007). Y. Abou Khalil is supported by a grant from Minist?re de l'Education Nationale et de la Technologie (France). J.T. Haas was supported by an EMBO Long Term Fellowship (ALTF277-2014). B. Staels is a recipient of an ERC Advanced Grant (no. 694717). Both are also supported by PreciNASH (ANR 16-RHUS-0006). Research at the Antwerp University Hospital was supported by the European Union: FP6 (HEPADIP Contract LSHMCT- 2005-018734). S. Francque has a senior clinical research fellowship from the Fund for Scientific Research (FWO) Flanders (1802154 N). S.E. Humphries received grants RG3008 and PG008/08 from the British Heart Foundation, and the support of the UCLH NIHR BRC. S.E. Humphries directs the UK Children's FH Register which has been supported by a grant from Pfizer (24052829) given by the International Atherosclerosis Society.

Publisher Copyright:
© 2021 American Heart Association, Inc.

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10.1161/CIRCRESAHA.120.318141Licens: CC BY

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Citationsformater

Zanoni, P., Panteloglou, G., Othman, A., Haas, J. T., Meier, R., Rimbert, A., Futema, M., Khalil, Y. A., 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., ... Von Eckardstein, A. (2022). Posttranscriptional Regulation of the Human LDL Receptor by the U2-Spliceosome. Circulation Research, 130(1), 80-95. https://doi.org/10.1161/CIRCRESAHA.120.318141

Zanoni, P, Panteloglou, G, Othman, A, Haas, JT, Meier, R, Rimbert, A, Futema, M, Khalil, YA, Norrelykke, SF, Rzepiela, AJ, Stoma, S, Stebler, M, Van Dijk, F, Wijers, M, Wolters, JC, Dalila, N, Huijkman, NCA, Smit, M, Gallo, A, Carreau, V, Philippi, A, Rabès, JP, Boileau, C, Visentin, M, Vonghia, L, Weyler, J, Francque, S, Verrijken, A, Verhaegen, A, Van Gaal, L, Van Der Graaf, A, Van Rosmalen, BV, Robert, J, Velagapudi, S, Yalcinkaya, M, Keel, M, Radosavljevic, S, Geier, A, Tybjaerg-Hansen, A, Varret, M, Rohrer, L, Humphries, SE, Staels, B, Van De Sluis, B, Kuivenhoven, JA & Von Eckardstein, A 2022, 'Posttranscriptional Regulation of the Human LDL Receptor by the U2-Spliceosome', Circulation Research, bind 130, nr. 1, s. 80-95. https://doi.org/10.1161/CIRCRESAHA.120.318141

@article{80ba810e1e804fb586161532e1482dba,

title = "Posttranscriptional Regulation of the Human LDL Receptor by the U2-Spliceosome",

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",

author = "Paolo Zanoni and Grigorios Panteloglou and Alaa Othman and Haas, {Joel T.} and Roger Meier and Antoine Rimbert and Marta Futema and Khalil, {Yara Abou} and Norrelykke, {Simon F.} and Rzepiela, {Andrzej J.} and Szymon Stoma and Michael Stebler and {Van Dijk}, Freerk and Melinde Wijers and Wolters, {Justina C.} and Nawar Dalila and Huijkman, {Nicolette C.A.} and Marieke Smit and Antonio Gallo and Val{\'e}rie Carreau and Anne Philippi and Rab{\`e}s, {Jean Pierre} and Catherine Boileau and Michele Visentin and Luisa Vonghia and Jonas Weyler and Sven Francque and An Verrijken and Ann Verhaegen and {Van Gaal}, Luc and {Van Der Graaf}, Adriaan and {Van Rosmalen}, {Belle V.} and Jerome Robert and Srividya Velagapudi and Mustafa Yalcinkaya and Michaela Keel and Silvija Radosavljevic and Andreas Geier and Anne Tybjaerg-Hansen and Mathilde Varret and Lucia Rohrer and Humphries, {Steve E.} and Bart Staels and {Van De Sluis}, Bart and Kuivenhoven, {Jan Albert} and {Von Eckardstein}, Arnold",

note = "Publisher Copyright: {\textcopyright} 2021 American Heart Association, Inc.",

year = "2022",

doi = "10.1161/CIRCRESAHA.120.318141",

language = "English",

volume = "130",

pages = "80--95",

journal = "Circulation Research",

issn = "0009-7330",

publisher = "AHA/ASA",

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TY - JOUR

T1 - Posttranscriptional Regulation of the Human LDL Receptor by the U2-Spliceosome

AU - Zanoni, Paolo

AU - Panteloglou, Grigorios

AU - Othman, Alaa

AU - Haas, Joel T.

AU - Meier, Roger

AU - Rimbert, Antoine

AU - Futema, Marta

AU - Khalil, Yara Abou

AU - Norrelykke, Simon F.

AU - Rzepiela, Andrzej J.

AU - Stoma, Szymon

AU - Stebler, Michael

AU - Van Dijk, Freerk

AU - Wijers, Melinde

AU - Wolters, Justina C.

AU - Dalila, Nawar

AU - Huijkman, Nicolette C.A.

AU - Smit, Marieke

AU - Gallo, Antonio

AU - Carreau, Valérie

AU - Philippi, Anne

AU - Rabès, Jean Pierre

AU - Boileau, Catherine

AU - Visentin, Michele

AU - Vonghia, Luisa

AU - Weyler, Jonas

AU - Francque, Sven

AU - Verrijken, An

AU - Verhaegen, Ann

AU - Van Gaal, Luc

AU - Van Der Graaf, Adriaan

AU - Van Rosmalen, Belle V.

AU - Robert, Jerome

AU - Velagapudi, Srividya

AU - Yalcinkaya, Mustafa

AU - Keel, Michaela

AU - Radosavljevic, Silvija

AU - Geier, Andreas

AU - Tybjaerg-Hansen, Anne

AU - Varret, Mathilde

AU - Rohrer, Lucia

AU - Humphries, Steve E.

AU - Staels, Bart

AU - Van De Sluis, Bart

AU - Kuivenhoven, Jan Albert

AU - Von Eckardstein, Arnold

PY - 2022

Y1 - 2022

N2 - 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.

AB - 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.

KW - Cardiovascular diseases

KW - Endocytosis

KW - Hepatocytes

KW - Hypercholesterolemia

KW - Spliceosomes

U2 - 10.1161/CIRCRESAHA.120.318141

DO - 10.1161/CIRCRESAHA.120.318141

M3 - Journal article

C2 - 34809444

AN - SCOPUS:85123207785

SN - 0009-7330

VL - 130

SP - 80

EP - 95

JO - Circulation Research

JF - Circulation Research

IS - 1

ER -