Functional interrogation of cellular Lp(a) uptake by genome-scale CRISPR screening
Functional interrogation of cellular Lp(a) uptake by genome-scale CRISPR screening
ABSTRACT An elevated level of lipoprotein(a), or Lp(a), in the bloodstream has been causally linked to the development of atherosclerotic cardiovascular disease and calcific aortic valve stenosis. Steady state levels of circulating lipoproteins are modulated by their rate of clearance, but the identity of the Lp(a) uptake receptor(s) has been controversial. In this study, we performed a genome-scale CRISPR screen to functionally interrogate all potential Lp(a) uptake regulators in HuH7 cells. Strikingly, the top positive and negative regulators of Lp(a) uptake in our screen were LDLR and MYLIP, encoding the LDL receptor and its ubiquitin ligase IDOL, respectively. We also found a significant correlation for other genes with established roles in LDLR regulation. No other gene products, including those previously proposed as Lp(a) receptors, exhibited a significant effect on Lp(a) uptake in our screen. We validated the functional influence of LDLR expression on HuH7 Lp(a) uptake, confirmed in vitro binding between the LDLR extracellular domain and purified Lp(a), and detected an association between loss-of-function LDLR variants and increased circulating Lp(a) levels in the UK Biobank cohort. Together, our findings support a central role for the LDL receptor in mediating Lp(a) uptake by hepatocytes.
Khan Taslima G.、Cunha Juliana Bragazzi、Raut Chinmay、Burroughs Michael、Smrcka Alan V.、Emmer Brian T.、Goonewardena Sascha N.、Speliotes Elizabeth K.
Program in Chemical Biology, University of MichiganDivision of Hospital Medicine, Department of Internal Medicine, University of MichiganDivision of Gastroenterology and Hepatology, Department of Internal Medicine, University of Michigan||Department of Computational Medicine and Bioinformatics, University of Michigan Medical SchoolDepartment of Pharmacology, University of MichiganDepartment of Pharmacology, University of MichiganDivision of Hospital Medicine, Department of Internal Medicine, University of Michigan||Frankel Cardiovascular Center, University of MichiganDivision of Cardiovascular Medicine, Department of Internal Medicine, University of Michigan||Frankel Cardiovascular Center, University of MichiganDivision of Gastroenterology and Hepatology, Department of Internal Medicine, University of Michigan||Department of Computational Medicine and Bioinformatics, University of Michigan Medical School
基础医学生物科学研究方法、生物科学研究技术分子生物学
Khan Taslima G.,Cunha Juliana Bragazzi,Raut Chinmay,Burroughs Michael,Smrcka Alan V.,Emmer Brian T.,Goonewardena Sascha N.,Speliotes Elizabeth K..Functional interrogation of cellular Lp(a) uptake by genome-scale CRISPR screening[EB/OL].(2025-03-28)[2025-05-18].https://www.biorxiv.org/content/10.1101/2024.05.11.593568.点此复制
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