title: Cholesteryl hemiesters alter lysosome structure and function and induce proinflammatory cytokine production in macrophages.
abstract: Cholesteryl hemiesters are oxidation products of polyunsaturated fatty acid esters of cholesterol. Their oxo-ester precursors have been identified as important components of the "core aldehydes" of human atheromata and in oxidized lipoproteins (Ox-LDL). We had previously shown, for the first time, that a single compound of this family, cholesteryl hemisuccinate (ChS), is sufficient to cause irreversible lysosomal lipid accumulation (lipidosis), and is toxic to macrophages. These features, coupled to others such as inflammation, are typically seen in atherosclerosis.
  
  
  
  To obtain insights into the mechanism of cholesteryl hemiester-induced pathological changes in lysosome function and induction of inflammation in vitro and assess their impact in vivo.
  
  
  
  We have examined the effects of ChS on macrophages (murine cell lines and primary cultures) in detail. Specifically, lysosomal morphology, pH, and proteolytic capacity were examined. Exposure of macrophages to sub-toxic ChS concentrations caused enlargement of the lysosomes, changes in their luminal pH, and accumulation of cargo in them. In primary mouse bone marrow-derived macrophages (BMDM), ChS-exposure increased the secretion of IL-1β, TNF-α and IL-6. In zebrafish larvae (wild-type AB and PU.1:EGFP), fed with a ChS-enriched diet, we observed lipid accumulation, myeloid cell-infiltration in their vasculature and decrease in larval survival. Under the same conditions the effects of ChS were more profound than the effects of free cholesterol (FC).
  
  
  
  Our data strongly suggest that cholesteryl hemiesters are pro-atherogenic lipids able to mimic features of Ox-LDL both in vitro and in vivo.
authors: CEDOC, NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, 1169-056 Lisboa, Portugal. Electronic address: neuzadomingues16@gmail.com.
authors: Neuza
authors: N
authors: Domingues
authors: CNC - Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal. Electronic address: estronca@ci.uc.pt.
authors: Luís M B B
authors: LMBB
authors: Estronca
authors: CNC - Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal. Electronic address: joao.silva@nms.unl.pt.
authors: João
authors: J
authors: Silva
authors: CEDOC, NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, 1169-056 Lisboa, Portugal. Electronic address: marisarego@gmail.com.
authors: Marisa R
authors: MR
authors: Encarnação
authors: CEDOC, NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, 1169-056 Lisboa, Portugal. Electronic address: Rita.Mateus@unige.ch.
authors: Rita
authors: R
authors: Mateus
authors: Life and Health Sciences Research Institute (ICVS), School of Health Sciences, University of Minho, Braga, Portugal; ICVS/3B's PT Government Associate Laboratory, Braga, Guimarães, Portugal. Electronic address: diogosilva@ecsaude.uminho.pt.
authors: Diogo
authors: D
authors: Silva
authors: CEDOC, NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, 1169-056 Lisboa, Portugal. Electronic address: isantarino@gmail.com.
authors: Inês B
authors: IB
authors: Santarino
authors: Life and Health Sciences Research Institute (ICVS), School of Health Sciences, University of Minho, Braga, Portugal; ICVS/3B's PT Government Associate Laboratory, Braga, Guimarães, Portugal. Electronic address: Margarida.Saraiva@ibmc.up.pt.
authors: Margarida
authors: M
authors: Saraiva
authors: CQC, Department of Chemistry, University of Coimbra, 3004-535 Coimbra, Portugal. Electronic address: misoares@ci.uc.pt.
authors: Maria I L
authors: MIL
authors: Soares
authors: CQC, Department of Chemistry, University of Coimbra, 3004-535 Coimbra, Portugal. Electronic address: tmelo@ci.uc.pt.
authors: Teresa M V D
authors: TMVD
authors: Pinho E Melo
authors: CEDOC, NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, 1169-056 Lisboa, Portugal. Electronic address: antonio.jacinto@nms.unl.pt.
authors: António
authors: A
authors: Jacinto
authors: CEDOC, NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, 1169-056 Lisboa, Portugal. Electronic address: winchil.vaz@nms.unl.pt.
authors: Winchil L C
authors: WLC
authors: Vaz
authors: CEDOC, NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, 1169-056 Lisboa, Portugal. Electronic address: otilia.vieira@nms.unl.pt.
authors: Otília V
authors: OV
authors: Vieira
mesh_terms: D000818:Animals; D050197:Atherosclerosis / Q000378:metabolism; D002460:Cell Line; D002784:Cholesterol / Q000378:metabolism*; D002788:Cholesterol Esters / Q000378:metabolism; D004952:Esters / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007249:Inflammation / Q000378:metabolism*; D053583:Interleukin-1beta / Q000378:metabolism*; D015850:Interleukin-6 / Q000378:metabolism*; D007814:Larva / Q000378:metabolism; D008064:Lipidoses / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D000067996:RAW 264.7 Cells; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*; D015027:Zebrafish
chemical_list: D002788:Cholesterol Esters; D004952:Esters; D053583:Interleukin-1beta; D015850:Interleukin-6; D014409:Tumor Necrosis Factor-alpha; D002784:Cholesterol; C013440:cholesteryl succinate
keywords: Atherosclerosis; Cholesteryl hemiesters; Inflammation; Lysosome malfunction; Oxidized lipids; Zebrafish larvae