Glucosylated cholesterol accumulates in atherosclerotic lesions and impacts macrophage immune response

Marques, A.R.A.; Ferreira, I.S.; Ribeiro, Q.; Guimaraes Da Lomba Ferraz, M.J.; Lopes, E.; Pinto, D.; ... ; Aerts, J.M.F.G. (2025)

Article / Letter to editor

Atherosclerosis can be described as a local acquired lysosomal storage disorder (LSD), resulting from the build-up of undegraded material in lysosomes. Atherosclerotic foam cells accumulate cholesterol (Chol) and glycosphingolipids (GSLs) within lysosomes. This constitutes the ideal milieu for the formation of a side product of lysosomal storage: glucosylated cholesterol (GlcChol), previously found in several LSDs. Using LC-MS/MS, we demonstrated that GlcChol is abundant in atherosclerotic lesions. Patients suffering from cardiovascular diseases presented unaltered plasma GlcChol levels but slightly elevated GlcChol/Chol ratios. Furthermore, we mimicked GlcChol formation in vitro by exposing macrophages (M phi) to a pro-atherogenic oxidized cholesteryl ester, an atherosclerosis foam cell model. Additionally, M phi exposed to GlcChol exhibited an enlarged and multi-nucleated phenotype. These M phi present signs of decreased proliferation and reduced pro-inflammatory capacity. Mechanistically, the process seems to be associated with activating the AMPK signaling pathway and the cyclin-dependent kinase inhibitor1 (CDKN1A/p21), in response to DNA damage inflicted by reactive oxygen species (ROS). At the organelle level, exposure to GlcChol impacted the lysosomal compartment, resulting in the activation of the mTOR signaling pathway and lysosomal biogenesis mediated by the transcription factor EB (TFEB). This suggests that high concentrations of GlcChol impact cellular homeostasis. In contrast, under this threshold, GlcChol formation most likely represents a relatively innocuous compensatory mechanism to cope with Chol and GSL build-up within lesions. Our findings demonstrate that glycosidase-mediated lipid modifications may play a role in the etiology of genetic and acquired LSDs, warranting further investigation.

• All authors: Marques, A.R.A.; Ferreira, I.S.; Ribeiro, Q.; Guimaraes Da Lomba Ferraz, M.J.; Lopes, E.; Pinto, D.; Hall, M.; Ramalho, J.; Artola Perez de Azanza, M.E.; Almeida, M.S.; Rodrigues, G.; Goncalves, P.A.; Ferreira, J.; Borbinha, C.; Marto, J.P.; Viana-Baptista, M.; Gouveiae e Melo, R.; Pedro, L.M.; Soares, M.I.L.; Vaz, W.L.C.; Vieira, O.V.; Aerts, J.M.F.G.
• Date: 2025-06-01
• Journal: Journal of Lipid Research
• Volume: 66
• Issue: 6
• DOI: doi:10.1016/j.jlr.2025.100825