title: Hemoglobinuria-related acute kidney injury is driven by intrarenal oxidative reactions triggering a heme toxicity response.
abstract: Intravascular hemolysis can result in hemoglobinuria with acute kidney injury. In this study we systematically explored two in vivo animal models and a related cell culture system to identify hemoglobinuria-triggered damage pathways. In models of stored blood transfusion and hemoglobin (Hb) exposure in guinea pigs and beagle dogs we found that hemoglobinuria led to intrarenal conversion of ferrous Hb(Fe(2+)) to ferric Hb(Fe(3+)), accumulation of free heme and Hb-cross-linking products, enhanced 4-hydroxynonenal reactivity in renal tissue, and acute tubule injury. These changes were associated in guinea pigs with activation of a renal cortex gene expression signature indicative of oxidative stress and activation of the unfolded protein response (UPR). Tubule cells of hemolytic animals demonstrated enhanced protein expression of heme oxygenase and heat shock protein and enhanced expression of acute kidney injury-related neutrophil gelatinase-associated lipocalin. These adverse changes were completely prevented by haptoglobin treatment. The in vivo findings were extrapolated to a MS-based proteome analysis of SILAC-labeled renal epithelial cells that were exposed to free heme within a concentration range estimate of renal tubule heme exposure. These experiments confirmed that free heme is a likely trigger of tubule barrier deregulation and oxidative cell damage and reinforced the hypothesis that uncontrolled free heme could trigger the UPR as an important pathway of renal injury during hemoglobinuria.
authors.affiliation: Division of Internal Medicine, University of Zürich, Zürich, Switzerland.
authors.forename: J W
authors.initials: JW
authors.lastname: Deuel
authors.affiliation: Division of Internal Medicine, University of Zürich, Zürich, Switzerland.|Institute of Anesthesiology, University of Zürich, Zürich, Switzerland.
authors.forename: C A
authors.initials: CA
authors.lastname: Schaer
authors.affiliation: Clinic for Small Animal Internal Medicine, University of Zürich, Zürich, Switzerland.
authors.forename: F S
authors.initials: FS
authors.lastname: Boretti
authors.affiliation: Functional Genomics Center Zürich, Swiss Federal Institute of Technology Zürich/University of Zürich, Zürich, Switzerland.
authors.forename: L
authors.initials: L
authors.lastname: Opitz
authors.affiliation: Laboratory of Physical Chemistry, ETH Zürich, Zürich, Switzerland.|Centro Universitario de la Defensa, Carretera de Huesca, Zaragoza, Spain.
authors.forename: I
authors.initials: I
authors.lastname: Garcia-Rubio
authors.affiliation: Laboratory of Biochemistry and Vascular Biology, Center of Biologics Evaluation and Research (CBER), FDA, Silver Spring, MA, USA.
authors.forename: J H
authors.initials: JH
authors.lastname: Baek
authors.affiliation: Institute of Anesthesiology, University of Zürich, Zürich, Switzerland.
authors.forename: D R
authors.initials: DR
authors.lastname: Spahn
authors.affiliation: Laboratory of Biochemistry and Vascular Biology, Center of Biologics Evaluation and Research (CBER), FDA, Silver Spring, MA, USA.
authors.forename: P W
authors.initials: PW
authors.lastname: Buehler
authors.affiliation: Division of Internal Medicine, University of Zürich, Zürich, Switzerland.|Institute of Evolutionary Medicine, University of Zürich, Zürich, Switzerland.
authors.forename: D J
authors.initials: DJ
authors.lastname: Schaer
mesh_terms: D058186:Acute Kidney Injury / Q000209:etiology* / Q000235:genetics; D000818:Animals; D004285:Dogs; D006168:Guinea Pigs; D006418:Heme; D006456:Hemoglobinuria / Q000209:etiology*; D006461:Hemolysis; D010084:Oxidation-Reduction; D018384:Oxidative Stress
chemical_list: D006418:Heme