Synchronized renal tubular cell death involves ferroptosis.
Receptor-interacting protein kinase 3 (RIPK3)-mediated necroptosis is thought to be the pathophysiologically predominant pathway that leads to regulated necrosis of parenchymal cells in ischemia-reperfusion injury (IRI), and loss of either Fas-associated protein with death domain (FADD) or caspase-8 is known to sensitize tissues to undergo spontaneous necroptosis. Here, we demonstrate that renal tubules do not undergo sensitization to necroptosis upon genetic ablation of either FADD or caspase-8 and that the RIPK1 inhibitor necrostatin-1 (Nec-1) does not protect freshly isolated tubules from hypoxic injury. In contrast, iron-dependent ferroptosis directly causes synchronized necrosis of renal tubules, as demonstrated by intravital microscopy in models of IRI and oxalate crystal-induced acute kidney injury. To suppress ferroptosis in vivo, we generated a novel third-generation ferrostatin (termed 16-86), which we demonstrate to be more stable, to metabolism and plasma, and more potent, compared with the first-in-class compound ferrostatin-1 (Fer-1). Even in conditions with extraordinarily severe IRI, 16-86 exerts strong protection to an extent which has not previously allowed survival in any murine setting. In addition, 16-86 further potentiates the strong protective effect on IRI mediated by combination therapy with necrostatins and compounds that inhibit mitochondrial permeability transition. Renal tubules thus represent a tissue that is not sensitized to necroptosis by loss of FADD or caspase-8. Finally, ferroptosis mediates postischemic and toxic renal necrosis, which may be therapeutically targeted by ferrostatins and by combination therapy.
Clinic for Nephrology and Hypertension, Christian-Albrechts-University Kiel, 24105 Kiel, Germany; andreas.linkermann@uksh.de krautwald@nephro.uni-kiel.de.
Andreas
http://orcid.org/0000-0001-6287-9725
A
Linkermann
Department of Biological Sciences and Department of Chemistry, University of Texas at El Paso, El Paso, TX 79902;
Rachid
R
Skouta
Department of Physiology, Christian-Albrechts-University Kiel, 24098 Kiel, Germany;
Nina
N
Himmerkus
Nephrologisches Zentrum, Medizinische Klinik und Poliklinik IV, Klinikum der Universität München, Ludwig-Maximilians-Universität München, 80366 Munich, Germany;
Shrikant R
SR
Mulay
Clinic for Nephrology and Hypertension, Christian-Albrechts-University Kiel, 24105 Kiel, Germany;
Christin
C
Dewitz
Clinic for Nephrology and Hypertension, Christian-Albrechts-University Kiel, 24105 Kiel, Germany;
Federica
F
De Zen
First Department of Pediatrics, Semmelweis University, H-1083 Budapest, Hungary;
Agnes
A
Prokai
Department of Otorhinolaryngology, Head and Neck Surgery, Ludwig-Maximilians-Universität München, 81366 Munich, Germany; Walter Brendel Centre of Experimental Medicine, Ludwig-Maximilians-Universität München, 81366 Munich, Germany;
Gabriele
G
Zuchtriegel
Department of Otorhinolaryngology, Head and Neck Surgery, Ludwig-Maximilians-Universität München, 81366 Munich, Germany; Walter Brendel Centre of Experimental Medicine, Ludwig-Maximilians-Universität München, 81366 Munich, Germany;
Fritz
F
Krombach
Institute for Research in Biomedicine, 08028 Barcelona, Spain; Institute for Genetics, University of Cologne, 50931 Cologne, Germany;
Patrick-Simon
PS
Welz
Department of Immunology, St. Jude Children's Research Hospital, Memphis, TN 38105-3678;
Ricardo
R
Weinlich
Molecular Signaling and Cell Death Unit, Inflammation Research Center, Vlaams Instituut voor Biotechnologie, Ghent University, 9052 Ghent, Belgium; Methusalem Program, Ghent University, 9052 Ghent, Belgium;
Tom
T
Vanden Berghe
Molecular Signaling and Cell Death Unit, Inflammation Research Center, Vlaams Instituut voor Biotechnologie, Ghent University, 9052 Ghent, Belgium; Methusalem Program, Ghent University, 9052 Ghent, Belgium;
Peter
P
Vandenabeele
Institute for Genetics, University of Cologne, 50931 Cologne, Germany;
Manolis
M
Pasparakis
Department of Physiology, Christian-Albrechts-University Kiel, 24098 Kiel, Germany;
Markus
M
Bleich
Division of Nephrology, Department of Internal Medicine, VA Healthcare System and University of Michigan, Ann Arbor, MI 48109-5676;
Joel M
JM
Weinberg
Department of Otorhinolaryngology, Head and Neck Surgery, Ludwig-Maximilians-Universität München, 81366 Munich, Germany; Walter Brendel Centre of Experimental Medicine, Ludwig-Maximilians-Universität München, 81366 Munich, Germany;
Christoph A
CA
Reichel
Department of Pathology, University of Hannover, 30625 Hannover, Germany;
Jan Hinrich
JH
Bräsen
Clinic for Nephrology and Hypertension, Christian-Albrechts-University Kiel, 24105 Kiel, Germany;
Ulrich
U
Kunzendorf
Nephrologisches Zentrum, Medizinische Klinik und Poliklinik IV, Klinikum der Universität München, Ludwig-Maximilians-Universität München, 80366 Munich, Germany;
Hans-Joachim
HJ
Anders
Department of Biological Sciences, Columbia University, New York, NY 10027; Department of Chemistry, Columbia University, New York, NY 10027; Howard Hughes Medical Institute, Columbia University, New York, NY 10027; and Department of Systems Biology, Columbia University Medical Center, New York, NY 10027.
Brent R
BR
Stockwell
Department of Immunology, St. Jude Children's Research Hospital, Memphis, TN 38105-3678;
Douglas R
DR
Green
Clinic for Nephrology and Hypertension, Christian-Albrechts-University Kiel, 24105 Kiel, Germany; andreas.linkermann@uksh.de krautwald@nephro.uni-kiel.de.
Stefan
S
Krautwald
D000818:Animals; D017209:Apoptosis; D001835:Body Weight; D053181:Caspase 8 / Q000235:genetics / Q000502:physiology; D053200:Fas-Associated Death Domain Protein / Q000235:genetics / Q000502:physiology; D007684:Kidney Tubules / Q000166:cytology*; D051379:Mice; D053422:Receptor-Interacting Protein Serine-Threonine Kinases / Q000235:genetics / Q000502:physiology; D015427:Reperfusion Injury / Q000517:prevention & control
C505534:Fadd protein, mouse; D053200:Fas-Associated Death Domain Protein; D053422:Receptor-Interacting Protein Serine-Threonine Kinases; C485615:Ripk3 protein, mouse; D053181:Caspase 8
apoptosis; ferroptosis; necroptosis; programmed cell death; regulated cell death