Awan, SaraLambert, MagalieImtiaz, AliAlpy, FabienTomasetto, CatherineOulad-Abdelghani, MustaphaSchaeffer, ChristineMoritz, ChloéJulien-David, DianeNajib, SouadMartinez, Laurent OMatz, Rachel LCollet, XavierSilva-Rojas, RobertoBöhm, JohannHerz, JoachimTerrand, JérômeBoucher, Philippe2025-01-272025-01-272022-01-21Circ Res. 2022 Jan 21;130(2):184-199.https://hdl.handle.net/20.500.12105/26137This work was supported by grants from Fédération Française de Cardiologie, and the Fondation de France (No. 00107044). R. Silva-Rojas was funded by Fondation Recherche Médicale doctoral fellowship (FRM, PLP20170939073). J. Herz was supported by grants from the National Heart, Lung, and Blood Institute (R37 HL063762), NIA (RF AG053391), the National Institute of Neurological Disorders and Stroke and NIA (R01 NS093382), BrightFocus A2016396S, the Bluefield Project to Cure FTD, and a Harrington Scholar Innovator Award (2019).Impairment of cellular cholesterol trafficking is at the heart of atherosclerotic lesions formation. This involves egress of cholesterol from the lysosomes and 2 lysosomal proteins, the NPC1 (Niemann-Pick C1) and NPC2 that promotes cholesterol trafficking. However, movement of cholesterol out the lysosome and how disrupted cholesterol trafficking leads to atherosclerosis is unclear. As the Wnt ligand, Wnt5a inhibits the intracellular accumulation of cholesterol in multiple cell types, we tested whether Wnt5a interacts with the lysosomal cholesterol export machinery and studied its role in atherosclerotic lesions formation. We generated mice deleted for the gene in vascular smooth muscle cells. To establish whether Wnt5a also protects against cholesterol accumulation in human vascular smooth muscle cells, we used a CRISPR/Cas9 guided nuclease approach to generate human vascular smooth muscle cells knockout for Wnt5a. We show that Wnt5a is a crucial component of the lysosomal cholesterol export machinery. By increasing lysosomal acid lipase expression, decreasing metabolic signaling by the mTORC1 (mechanistic target of rapamycin complex 1) kinase, and through binding to NPC1 and NPC2, Wnt5a senses changes in dietary cholesterol supply and promotes lysosomal cholesterol egress to the endoplasmic reticulum. Consequently, loss of Wnt5a decoupled mTORC1 from variations in lysosomal sterol levels, disrupted lysosomal function, decreased cholesterol content in the endoplasmic reticulum, and promoted atherosclerosis. These results reveal an unexpected function of the Wnt5a pathway as essential for maintaining cholesterol homeostasis in vivo.engVoRhttp://creativecommons.org/licenses/by-nc-nd/4.0/atherosclerosishomeostasisligandslysosomesAttribution-NonCommercial-NoDerivatives 4.0 International34886684130(2)184-199open access