title: Inhibition of proteasome by bortezomib causes intracellular aggregation of hepatic serpins and increases the latent circulating form of antithrombin.
abstract: Conformational diseases include heterogeneous disorders sharing a similar pathological mechanism, leading to intracellular aggregation of proteins with toxic effects. Serpins are commonly involved in these diseases. These are structurally sensitive molecules that modify their folding under even minor genetic or environmental variations. Indeed, under normal conditions, the rate of misfolding of serpins is high and unfolded serpins must be degraded by the proteasome system. Our aim was to study the effects of bortezomib, a proteasome inhibitor, on conformationally sensitive serpins. The effects of bortezomib were analysed in patients with multiple myeloma, HepG2 cells, and Swiss mice, as well as in vitro. Levels, anti-FXa activity, heparin affinity, and conformational features of antithrombin, a relevant anticoagulant serpin, were analysed. Histological, ultrastructural features and immunohistological distribution of antithrombin and alpha1-antitrypsin (another hepatic serpin) were evaluated. We also studied the intracellular accumulation of conformationally sensitive (fibrinogen) or non-sensitive (prothrombin) hepatic proteins. The inhibition of the proteasome caused intracellular accumulation and aggregation of serpins within the endoplasmic reticulum that was associated with confronting cisternae and Mallory body formation. These effects were accompanied by a heat stress response. Bortezomib also increased the levels of intracellular fibrinogen, but has no significant effect on prothrombin. Finally, bortezomib had only minor effects on the mature circulating antithrombin, with increased amounts of latent antithrombin in plasma. These results suggest that the impairment of proteasomal activities leads to an intracellular accumulation of conformationally sensitive proteins and might facilitate the release of misfolded serpins into circulation where they adopt more stable conformations.
authors.affiliation: Department of Medicine, Centro Regional de Hemodonación, Murcia, Spain.
authors.forename: David
authors.initials: D
authors.lastname: Hernández-Espinosa
authors.forename: Antonia
authors.initials: A
authors.lastname: Miñano
authors.forename: Constantino
authors.initials: C
authors.lastname: Martínez
authors.forename: Adriana
authors.initials: A
authors.lastname: Ordoñez
authors.forename: Elena
authors.initials: E
authors.lastname: Pérez-Ceballos
authors.forename: Felipe
authors.initials: F
authors.lastname: de Arriba
authors.forename: Rubén A
authors.initials: RA
authors.lastname: Mota
authors.forename: Francisca
authors.initials: F
authors.lastname: Ferrer
authors.forename: Marcos
authors.initials: M
authors.lastname: González
authors.forename: Vicente
authors.initials: V
authors.lastname: Vicente
authors.forename: Javier
authors.initials: J
authors.lastname: Corral
mesh_terms: D000483:Alleles; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D000991:Antithrombins / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D001897:Boronic Acids / Q000008:administration & dosage / Q000494:pharmacology*; D000069286:Bortezomib; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy / Q000473:pathology; D016022:Case-Control Studies; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D004334:Drug Administration Schedule; D004353:Drug Evaluation, Preclinical; D004721:Endoplasmic Reticulum / Q000378:metabolism; D065427:Factor Xa Inhibitors; D005340:Fibrinogen / Q000096:biosynthesis; D006801:Humans; D007150:Immunohistochemistry; D007275:Injections, Intravenous; D019272:Leukocyte Elastase / Q000009:adverse effects / Q000097:blood / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure; D008099:Liver / Q000378:metabolism* / Q000473:pathology / Q000648:ultrastructure; D008113:Liver Neoplasms / Q000188:drug therapy / Q000473:pathology; D008297:Male; D051379:Mice; D018832:Molecular Chaperones / Q000378:metabolism; D009101:Multiple Myeloma / Q000188:drug therapy / Q000473:pathology; D061988:Proteasome Inhibitors; D011506:Proteins / Q000378:metabolism; D011719:Pyrazines / Q000008:administration & dosage / Q000494:pharmacology*; D015843:Serpins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D025801:Ubiquitin / Q000378:metabolism; D000515:alpha 1-Antitrypsin / Q000009:adverse effects / Q000097:blood / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D000970:Antineoplastic Agents; D000991:Antithrombins; D001897:Boronic Acids; D065427:Factor Xa Inhibitors; C488922:Mallory body protein, mouse; D018832:Molecular Chaperones; D061988:Proteasome Inhibitors; D011506:Proteins; D011719:Pyrazines; D015843:Serpins; D025801:Ubiquitin; D000515:alpha 1-Antitrypsin; C037741:alpha 1-antitrypsin-leukocyte elastase complex; D000069286:Bortezomib; D005340:Fibrinogen; D019272:Leukocyte Elastase