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 Flyvbjerg Nørrelykke; Andrzej Rzepiela; Szymon Stoma; Michael Stebler; Freerk van Dijk; Melinde Wijers; Justina C Wolters; Nawar Dalila; Nicolette Huijkman; Marieke Smit; Antonio Gallo; Valerie Carreau; Anne Philippi; Jean-Pierre Rabès; Catherine Boileau; Michele Visentin; Luisa Vonghia; Jonas Weyler; Sven Francque; An Verrijken; Ann Verhaegen; Luc F Van Gaal; Adriaan van der Graaf; Belle V van Rosmalen; Jérôme Robert; Srividya Velagapudi; Mustafa Yalcinkaya; Michaela Keel; Silvija Radosavljevic; Andreas Geier; Anne Tybjærg-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 Flyvbjerg Nørrelykke, Andrzej Rzepiela, Szymon Stoma, Michael Stebler, Freerk van Dijk, Melinde Wijers, Justina C Wolters, Nawar Dalila, Nicolette Huijkman, Marieke Smit, Antonio Gallo, Valerie CarreauAnne Philippi, Jean-Pierre Rabès, Catherine Boileau, Michele Visentin, Luisa Vonghia, Jonas Weyler, Sven Francque, An Verrijken, Ann Verhaegen, Luc F Van Gaal, Adriaan van der Graaf, Belle V van Rosmalen, Jérôme Robert, Srividya Velagapudi, Mustafa Yalcinkaya, Michaela Keel, Silvija Radosavljevic, Andreas Geier, Anne Tybjærg-Hansen, Mathilde Varret, Lucia Rohrer, Steve E Humphries, Bart Staels, Bart van de Sluis, Jan Albert Kuivenhoven, Arnold Von Eckardstein^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

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Abstract

Background: The low-density lipoprotein receptor (LDLR) 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 datasets 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 non-transfected cells nor in human plasma. The hepatic expression of the intron 3 retention transcript is increased in non-alcoholic fatty liver disease as well as after bariatric surgery. Its expression in blood cells correlates with LDL-cholesterol and age. Single nucleotide polymorphisms and three rare variants of one spliceosome gene, RBM25, are associated with LDL-cholesterol in the population and familial hypercholesterolemia, respectively. Compared to overexpression of wild type RBM25, overexpression of the three rare RBM25 mutants in Huh-7 cells led to lower LDL uptake.

Conclusions: We identified a novel mechanism of post-transcriptional regulation of LDLR activity in humans and associations of genetic variants of RBM25 with LDL-cholesterol levels.

Original language	English
Pages (from-to)	80-95
Journal	Circulation Research
Volume	130
Issue number	1
Early online date	23-Nov-2021
DOIs	https://doi.org/10.1161/CIRCRESAHA.120.318141
Publication status	Published - 7-Jan-2022

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Zanoni, P., Panteloglou, G., Othman, A., Haas, J. T., Meier, R., Rimbert, A., Futema, M., Abou-Khalil, Y., Nørrelykke, S. F., Rzepiela, A., Stoma, S., Stebler, M., Dijk, F. V., Wijers, M., Wolters, J. C., Dalila, N., Huijkman, N., 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

@article{c93ba9907afb46da9fcb24a8f7404a9b,

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

abstract = "Background: The low-density lipoprotein receptor (LDLR) 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 datasets 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 non-transfected cells nor in human plasma. The hepatic expression of the intron 3 retention transcript is increased in non-alcoholic fatty liver disease as well as after bariatric surgery. Its expression in blood cells correlates with LDL-cholesterol and age. Single nucleotide polymorphisms and three rare variants of one spliceosome gene, RBM25, are associated with LDL-cholesterol in the population and familial hypercholesterolemia, respectively. Compared to overexpression of wild type RBM25, overexpression of the three rare RBM25 mutants in Huh-7 cells led to lower LDL uptake.Conclusions: We identified a novel mechanism of post-transcriptional regulation of LDLR activity in humans and associations of genetic variants of RBM25 with LDL-cholesterol levels.",

author = "Paolo Zanoni and Grigorios Panteloglou and Alaa Othman and Haas, {Joel T} and Roger Meier and Antoine Rimbert and Marta Futema and Yara Abou-Khalil and N{\o}rrelykke, {Simon Flyvbjerg} and Andrzej Rzepiela and Szymon Stoma and Michael Stebler and Dijk, {Freerk van} and Melinde Wijers and Wolters, {Justina C} and Nawar Dalila and Nicolette Huijkman and Marieke Smit and Antonio Gallo and Valerie Carreau and Anne Philippi and Jean-Pierre Rab{\`e}s 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 F} and {van der Graaf}, Adriaan and {van Rosmalen}, {Belle V} and J{\'e}r{\^o}me Robert and Srividya Velagapudi and Mustafa Yalcinkaya and Michaela Keel and Silvija Radosavljevic and Andreas Geier and Anne Tybj{\ae}rg-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",

year = "2022",

month = jan,

day = "7",

doi = "10.1161/CIRCRESAHA.120.318141",

language = "English",

volume = "130",

pages = "80--95",

journal = "Circulation Research",

issn = "0009-7330",

publisher = "LIPPINCOTT WILLIAMS & WILKINS",

number = "1",

}

Zanoni, P, Panteloglou, G, Othman, A, Haas, JT, Meier, R, Rimbert, A, Futema, M, Abou-Khalil, Y, Nørrelykke, SF, Rzepiela, A, Stoma, S, Stebler, M, Dijk, FV , Wijers, M , Wolters, JC, Dalila, N, Huijkman, N , 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, LF, van der Graaf, A, van Rosmalen, BV, Robert, J, Velagapudi, S, Yalcinkaya, M, Keel, M, Radosavljevic, S, Geier, A, Tybjærg-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, vol. 130, no. 1, pp. 80-95. https://doi.org/10.1161/CIRCRESAHA.120.318141

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 - Abou-Khalil, Yara

AU - Nørrelykke, Simon Flyvbjerg

AU - Rzepiela, Andrzej

AU - Stoma, Szymon

AU - Stebler, Michael

AU - Dijk, Freerk van

AU - Wijers, Melinde

AU - Wolters, Justina C

AU - Dalila, Nawar

AU - Huijkman, Nicolette

AU - Smit, Marieke

AU - Gallo, Antonio

AU - Carreau, Valerie

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 F

AU - van der Graaf, Adriaan

AU - van Rosmalen, Belle V

AU - Robert, Jérôme

AU - Velagapudi, Srividya

AU - Yalcinkaya, Mustafa

AU - Keel, Michaela

AU - Radosavljevic, Silvija

AU - Geier, Andreas

AU - Tybjærg-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 - 2022/1/7

Y1 - 2022/1/7

N2 - Background: The low-density lipoprotein receptor (LDLR) 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 datasets 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 non-transfected cells nor in human plasma. The hepatic expression of the intron 3 retention transcript is increased in non-alcoholic fatty liver disease as well as after bariatric surgery. Its expression in blood cells correlates with LDL-cholesterol and age. Single nucleotide polymorphisms and three rare variants of one spliceosome gene, RBM25, are associated with LDL-cholesterol in the population and familial hypercholesterolemia, respectively. Compared to overexpression of wild type RBM25, overexpression of the three rare RBM25 mutants in Huh-7 cells led to lower LDL uptake.Conclusions: We identified a novel mechanism of post-transcriptional regulation of LDLR activity in humans and associations of genetic variants of RBM25 with LDL-cholesterol levels.

AB - Background: The low-density lipoprotein receptor (LDLR) 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 datasets 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 non-transfected cells nor in human plasma. The hepatic expression of the intron 3 retention transcript is increased in non-alcoholic fatty liver disease as well as after bariatric surgery. Its expression in blood cells correlates with LDL-cholesterol and age. Single nucleotide polymorphisms and three rare variants of one spliceosome gene, RBM25, are associated with LDL-cholesterol in the population and familial hypercholesterolemia, respectively. Compared to overexpression of wild type RBM25, overexpression of the three rare RBM25 mutants in Huh-7 cells led to lower LDL uptake.Conclusions: We identified a novel mechanism of post-transcriptional regulation of LDLR activity in humans and associations of genetic variants of RBM25 with LDL-cholesterol levels.

U2 - 10.1161/CIRCRESAHA.120.318141

DO - 10.1161/CIRCRESAHA.120.318141

M3 - Article

C2 - 34809444

SN - 0009-7330

VL - 130

SP - 80

EP - 95

JO - Circulation Research

JF - Circulation Research

IS - 1

ER -

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

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