title: Clozapine-induced mitochondria alterations and inflammation in brain and insulin-responsive cells.
abstract: BACKGROUND
  Metabolic syndrome (MetS) is a constellation of factors including abdominal obesity, hyperglycemia, dyslipidemias, and hypertension that increase morbidity and mortality from diabetes and cardiovascular diseases and affects more than a third of the population in the US. Clozapine, an atypical antipsychotic used for the treatment of schizophrenia, has been found to cause drug-induced metabolic syndrome (DIMS) and may be a useful tool for studying cellular and molecular changes associated with MetS and DIMS. Mitochondria dysfunction, oxidative stress and inflammation are mechanisms proposed for the development of clozapine-related DIMS. In this study, the effects of clozapine on mitochondrial function and inflammation in insulin responsive and obesity-associated cultured cell lines were examined.
  
  
  RESULTS
  Cultured mouse myoblasts (C2C12), adipocytes (3T3-L1), hepatocytes (FL-83B), and monocytes (RAW 264.7) were treated with 0, 25, 50 and 75 µM clozapine for 24 hours. The mitochondrial selective probe TMRM was used to assess membrane potential and morphology. ATP levels from cell lysates were determined by bioluminescence assay. Cytokine levels in cell supernatants were assessed using a multiplex array. Clozapine was found to alter mitochondria morphology, membrane potential, and volume, and reduce ATP levels in all cell lines. Clozapine also significantly induced the production of proinflammatory cytokines IL-6, GM-CSF and IL12-p70, and this response was particularly robust in the monocyte cell line.
  
  
  CONCLUSIONS
  Clozapine damages mitochondria and promotes inflammation in insulin responsive cells and obesity-associated cell types. These phenomena are closely associated with changes observed in human and animal studies of MetS, obesity, insulin resistance, and diabetes. Therefore, the use of clozapine in DIMS may be an important and relevant tool for investigating cellular and molecular changes associated with the development of these diseases in the general population.
authors.affiliation: Department of Biological Sciences, Saint Mary's University, San Antonio, Texas, United States of America.
authors.forename: Verόnica
authors.initials: V
authors.lastname: Contreras-Shannon
authors.forename: Dylan L
authors.initials: DL
authors.lastname: Heart
authors.forename: R Madelaine
authors.initials: RM
authors.lastname: Paredes
authors.forename: Erica
authors.initials: E
authors.lastname: Navaira
authors.forename: Gabriel
authors.initials: G
authors.lastname: Catano
authors.forename: Shivani Kaushal
authors.initials: SK
authors.lastname: Maffi
authors.forename: Consuelo
authors.initials: C
authors.lastname: Walss-Bass
mesh_terms: D041721:3T3-L1 Cells; D000255:Adenosine Triphosphate / Q000096:biosynthesis; D017667:Adipocytes / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D014150:Antipsychotic Agents / Q000009:adverse effects*; D001921:Brain / Q000378:metabolism / Q000473:pathology; D002460:Cell Line; D003024:Clozapine / Q000009:adverse effects*; D016207:Cytokines / Q000096:biosynthesis; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism; D007249:Inflammation / Q000378:metabolism; D018836:Inflammation Mediators / Q000378:metabolism; D007328:Insulin / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D024821:Metabolic Syndrome; D051379:Mice; D008928:Mitochondria / Q000187:drug effects*; D032446:Myoblasts / Q000187:drug effects / Q000378:metabolism; D009447:Neuroblastoma / Q000378:metabolism
chemical_list: D014150:Antipsychotic Agents; D016207:Cytokines; D018836:Inflammation Mediators; D007328:Insulin; D000255:Adenosine Triphosphate; D003024:Clozapine