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

Onderzoeksoutput: Bijdrage aan tijdschriftArtikelpeer review

Samenvatting

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.

Originele taal-2Engels
Pagina's (van-tot)80-95
Aantal pagina's16
TijdschriftCirculation Research
Volume130
Nummer van het tijdschrift1
DOI's
StatusGepubliceerd - 7 jan. 2022
Extern gepubliceerdJa

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