title: Endothelin-1 receptor antagonists regulate cell surface-associated protein disulfide isomerase in sickle cell disease.
abstract: Increased endothelin-1 (ET-1) levels, disordered thiol protein status, and erythrocyte hydration status play important roles in sickle cell disease (SCD) through unresolved mechanisms. Protein disulfide isomerase (PDI) is an oxidoreductase that mediates thiol/disulfide interchange reactions. We provide evidence that PDI is present in human and mouse erythrocyte membranes and that selective blockade with monoclonal antibodies against PDI leads to reduced Gardos channel activity (1.6±0.03 to 0.56±0.02 mmol·10(13) cell(-1)·min(-1), P<0.001) and density of sickle erythrocytes (D50: 1.115±0.001 to 1.104±0.001 g/ml, P=0.012) with an IC50 of 4 ng/ml. We observed that erythrocyte associated-PDI activity was increased in the presence of ET-1 (3.1±0.2 to 5.6±0.4%, P<0.0001) through a mechanism that includes casein kinase II. Consistent with these results, in vivo treatment of BERK sickle transgenic mice with ET-1 receptor antagonists lowered circulating and erythrocyte associated-PDI activity (7.1±0.3 to 5.2±0.2%, P<0.0001) while improving hematological parameters and Gardos channel activity. Thus, our results suggest that PDI is a novel target in SCD that regulates erythrocyte volume and oxidative stress and may contribute to cellular adhesion and endothelial activation leading to vasoocclusion as observed in SCD.
authors.affiliation: 1Department of Laboratory Medicine, Bader 7, Boston Children's Hospital, 300 Longwood Ave., Boston, MA 02115, USA. alicia.rivera@childrens.harvard.edu.
authors.forename: Gregory N
authors.initials: GN
authors.lastname: Prado
authors.forename: Jose R
authors.initials: JR
authors.lastname: Romero
authors.forename: Alicia
authors.initials: A
authors.lastname: Rivera
mesh_terms: D000755:Anemia, Sickle Cell / Q000097:blood / Q000201:enzymology* / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D061207:Calcium Ionophores / Q000494:pharmacology; D047390:Casein Kinase II / Q000037:antagonists & inhibitors / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D065130:Endothelin A Receptor Antagonists; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004911:Erythrocyte Volume / Q000187:drug effects; D004912:Erythrocytes / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D006801:Humans; D051660:Intermediate-Conductance Calcium-Activated Potassium Channels / Q000037:antagonists & inhibitors / Q000378:metabolism; D051379:Mice; D018384:Oxidative Stress; D019704:Protein Disulfide-Isomerases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000494:pharmacology
chemical_list: D000911:Antibodies, Monoclonal; D061207:Calcium Ionophores; D065130:Endothelin A Receptor Antagonists; D004791:Enzyme Inhibitors; D051660:Intermediate-Conductance Calcium-Activated Potassium Channels; C496923:KCNN4 protein, human; C496925:Kcnn4 protein, mouse; D013438:Sulfhydryl Compounds; D000001:Calcimycin; D047390:Casein Kinase II; D019704:Protein Disulfide-Isomerases
keywords: Berkeley transgenic SCD mice; Gardos channel; erythrocytes; redox reactions
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