title: Proteomic analysis of phospholipidosis in citalopram treated U937 cells--support for the cholesterol biosynthesis hypothesis.
abstract: Excessive accumulation of phospholipids leads to phospholipidosis (PL), which disrupts cellular functions, in extreme cases leading to acute or chronic disease. Citalopram and many other cationic amphiphilic drugs (CADs) have been shown to cause PL both in vitro and in vivo. Recent toxicogenomic studies suggest four hypothetical mechanisms for PL (lysosomal enzyme transport decrease, lysosomal phospholipase activity decrease, phospholipids biosynthesis increase or cholesterol biosynthesis increase). However, the post-genomic steps remain largely unknown and proteomic analyses hold significant promise for defining mechanisms of PL induction. In this study U937 monoblastoid cells were exposed to citalopram hydrobromide for 24 h (0, 20, 100 or 200 microM as citalopram free base) and then harvested for whole cell proteomic analysis using 2-D gel electrophoresis, or transmission electron microscopy (TEM). Protein spots that were significantly altered versus controls were analysed by MALDI-TOF mass spectrometry. Up-regulated proteins were Glyoxalase-I (Glo 1) and 3-Hydroxy-3-methylglutaryl-Coenzyme A synthase 1 (HMGCS1) in cells with PL shown by TEM (favouring the cholesterol biosynthesis increase hypothesis for citalopram induced PL). Other altered proteins were catalase (up-regulated), beta-actin (up-regulated) and 14-3-3 protein (down-regulated). The function of several of the successfully identified proteins indicates a potential perturbed lipid metabolism.
authors.affiliation: AstraZeneca R&D, Safety Assessment, S-151 85 Södertälje, Sweden.
authors.forename: Thomas H
authors.initials: TH
authors.lastname: Hutchinson
authors.forename: Yilmaz
authors.initials: Y
authors.lastname: Mahshid
authors.forename: Raija
authors.initials: R
authors.lastname: Jönsson
authors.forename: Christina
authors.initials: C
authors.lastname: Björklund
authors.forename: Kerstin
authors.initials: K
authors.lastname: Kenne
mesh_terms: D002470:Cell Survival / Q000187:drug effects; D002784:Cholesterol / Q000096:biosynthesis*; D015283:Citalopram / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D006904:Hydroxymethylglutaryl-CoA Synthase / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D017367:Selective Serotonin Reuptake Inhibitors / Q000494:pharmacology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D020298:U937 Cells / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D015854:Up-Regulation
chemical_list: D002784:Cholesterol; D015283:Citalopram; D006904:Hydroxymethylglutaryl-CoA Synthase; D010743:Phospholipids; D017367:Selective Serotonin Reuptake Inhibitors