Dysfunction of the key ferroptosis-surveilling systems hypersensitizes mice to tubular necrosis during acute kidney injury

Wulf Tonnus; Claudia Meyer; Christian Steinebach; Alexia Belavgeni; Anne von Mässenhausen; Nadia Zamora Gonzalez; Francesca Maremonti; Florian Gembardt; Nina Himmerkus; Markus Latk; Sophie Locke; Julian Marschner; Wenjun Li; Spencer Short; Sebastian Doll; Irina Ingold; Bettina Proneth; Christoph Daniel; Nazanin Kabgani; Rafael Kramann; Stephen Motika; Paul J Hergenrother; Stefan R Bornstein; Christian Hugo; Jan Ulrich Becker; Kerstin Amann; Hans-Joachim Anders; Daniel Kreisel; Derek Pratt; Michael Gütschow; Marcus Conrad; Andreas Linkermann

doi:10.1038/s41467-021-24712-6

Dysfunction of the key ferroptosis-surveilling systems hypersensitizes mice to tubular necrosis during acute kidney injury

Nat Commun. 2021 Jul 20;12(1):4402. doi: 10.1038/s41467-021-24712-6.

Authors

Wulf Tonnus^#^{1

2}, Claudia Meyer^#^{1

2}, Christian Steinebach³, Alexia Belavgeni^{1

2}, Anne von Mässenhausen^{1

2}, Nadia Zamora Gonzalez^{1

2}, Francesca Maremonti^{1

2}, Florian Gembardt¹, Nina Himmerkus⁴, Markus Latk^{1

2}, Sophie Locke^{1

2}, Julian Marschner⁵, Wenjun Li⁶, Spencer Short⁷, Sebastian Doll⁸, Irina Ingold⁸, Bettina Proneth⁸, Christoph Daniel⁹, Nazanin Kabgani¹⁰, Rafael Kramann^{10

11}, Stephen Motika¹², Paul J Hergenrother¹², Stefan R Bornstein^{13

14

15

16

17}, Christian Hugo¹, Jan Ulrich Becker¹⁸, Kerstin Amann⁹, Hans-Joachim Anders⁵, Daniel Kreisel^{6

19}, Derek Pratt⁷, Michael Gütschow³, Marcus Conrad^{8

20}, Andreas Linkermann^{21

22}

Affiliations

¹ Division of Nephrology, Department of Internal Medicine 3, University Hospital Carl Gustav Carus at the Technische Universität Dresden, Dresden, Germany.
² Biotechnology Center, Technische Universität Dresden, Dresden, Germany.
³ Pharmaceutical Institute, Pharmaceutical Chemistry I, University of Bonn, Bonn, Germany.
⁴ Institute of Physiology, Christian-Albrecht-University Kiel, Kiel, Germany.
⁵ Division of Nephrology, Department of Medicine IV, University Hospital LMU Munich, Munich, Germany.
⁶ Department of Surgery, Washington University, Saint Louis, MO, USA.
⁷ Department of Chemistry and Biomolecular Sciences, University of Ottawa, Ottawa, ON, Canada.
⁸ Institute of Metabolism and Cell Death, Helmholtz Zentrum München, Neuherberg, Germany.
⁹ Department of Nephropathology, Friedrich-Alexander University (FAU) Erlangen-Nürnberg, Erlangen, Germany.
¹⁰ Clinic for Renal and Hypertensive Disorders, Rheumatological and Immunological Disease, University Hospital of the RWTH Aachen, Aachen, Germany.
¹¹ Department of Internal Medicine, Nephrology and Transplantation, Erasmus Medical Center, Rotterdam, The Netherlands.
¹² Department of Pathobiology, University of Illinois, Urbana, IL, USA.
¹³ Department of Internal Medicine 3, University Hospital Carl Gustav Carus at the Technische Universität Dresden, Dresden, Germany.
¹⁴ Diabetes and Nutritional Sciences, King's College London, London, UK.
¹⁵ Center for Regenerative Therapies, Technische Universität Dresden, Dresden, Germany.
¹⁶ Paul Langerhans Institute Dresden of Helmholtz Centre Munich at University Clinic Carl Gustav Carus of TU Dresden Faculty of Medicine, Dresden, Germany.
¹⁷ Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore City, Singapore.
¹⁸ Institute of Pathology, University Hospital of Cologne, Cologne, Germany.
¹⁹ Department of Pathology and Immunology, Washington University, Saint Louis, MO, USA.
²⁰ National Research Medical University, Laboratory of Experimental Oncology, Moscow, Russia.
²¹ Division of Nephrology, Department of Internal Medicine 3, University Hospital Carl Gustav Carus at the Technische Universität Dresden, Dresden, Germany. andreas.linkermann@ukdd.de.
²² Biotechnology Center, Technische Universität Dresden, Dresden, Germany. andreas.linkermann@ukdd.de.

^# Contributed equally.

Abstract

Acute kidney injury (AKI) is morphologically characterized by a synchronized plasma membrane rupture of cells in a specific section of a nephron, referred to as acute tubular necrosis (ATN). Whereas the involvement of necroptosis is well characterized, genetic evidence supporting the contribution of ferroptosis is lacking. Here, we demonstrate that the loss of ferroptosis suppressor protein 1 (Fsp1) or the targeted manipulation of the active center of the selenoprotein glutathione peroxidase 4 (Gpx4^cys/-) sensitize kidneys to tubular ferroptosis, resulting in a unique morphological pattern of tubular necrosis. Given the unmet medical need to clinically inhibit AKI, we generated a combined small molecule inhibitor (Nec-1f) that simultaneously targets receptor interacting protein kinase 1 (RIPK1) and ferroptosis in cell lines, in freshly isolated primary kidney tubules and in mouse models of cardiac transplantation and of AKI and improved survival in models of ischemia-reperfusion injury. Based on genetic and pharmacological evidence, we conclude that GPX4 dysfunction hypersensitizes mice to ATN during AKI. Additionally, we introduce Nec-1f, a solid inhibitor of RIPK1 and weak inhibitor of ferroptosis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Acute Kidney Injury / drug therapy
Acute Kidney Injury / etiology
Acute Kidney Injury / pathology*
Animals
Apoptosis Regulatory Proteins / genetics
Apoptosis Regulatory Proteins / metabolism
Cisplatin / administration & dosage
Cisplatin / toxicity
Disease Models, Animal
Epithelial Cells
Female
Ferroptosis / drug effects
Ferroptosis / physiology*
Gene Knockdown Techniques
HT29 Cells
Heart Transplantation / adverse effects
Humans
Imidazoles / chemistry
Imidazoles / pharmacology
Imidazoles / therapeutic use
Indoles / chemistry
Indoles / pharmacology
Indoles / therapeutic use
Kidney Tubules / pathology*
Male
Mice
Mice, Transgenic
Microsomes, Liver
Mitochondrial Proteins / metabolism
NIH 3T3 Cells
Necrosis / drug therapy
Necrosis / etiology
Necrosis / pathology
Oxidoreductases / genetics
Oxidoreductases / metabolism
Phospholipid Hydroperoxide Glutathione Peroxidase / antagonists & inhibitors
Phospholipid Hydroperoxide Glutathione Peroxidase / metabolism
Primary Cell Culture
Receptor-Interacting Protein Serine-Threonine Kinases / antagonists & inhibitors
Receptor-Interacting Protein Serine-Threonine Kinases / metabolism
Reperfusion Injury / drug therapy
Reperfusion Injury / etiology
Reperfusion Injury / pathology*

Substances

AIFM3 protein, human
Apoptosis Regulatory Proteins
Imidazoles
Indoles
Mitochondrial Proteins
necrostatin-1
ferroptosis suppressor protein 1, mouse
Oxidoreductases
Phospholipid Hydroperoxide Glutathione Peroxidase
glutathione peroxidase 4, mouse
Receptor-Interacting Protein Serine-Threonine Kinases
Ripk1 protein, mouse
Cisplatin