RIP kinase 1–dependent endothelial necroptosis underlies systemic inflammatory response syndrome
Matija Zelic, … , Manolis Pasparakis, Michelle A. Kelliher
Matija Zelic, … , Manolis Pasparakis, Michelle A. Kelliher
Published April 16, 2018
Citation Information: J Clin Invest. 2018;128(5):2064-2075. https://doi.org/10.1172/JCI96147.
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Research Article Immunology Inflammation

RIP kinase 1–dependent endothelial necroptosis underlies systemic inflammatory response syndrome

Abstract

Receptor interacting protein kinase 1 (RIPK1) has important kinase-dependent and kinase-independent scaffolding functions that activate or prevent apoptosis or necroptosis in a cell context–dependent manner. The kinase activity of RIPK1 mediates hypothermia and lethality in a mouse model of TNF-induced shock, reflecting the hyperinflammatory state of systemic inflammatory response syndrome (SIRS), where the proinflammatory “cytokine storm” has long been viewed as detrimental. Here, we demonstrate that cytokine and chemokine levels did not predict survival and, importantly, that kinase-inactive Ripk1D138N/D138N hematopoietic cells afforded little protection from TNF- or TNF/zVAD-induced shock in reconstituted mice. Unexpectedly, RIPK1 kinase–inactive mice transplanted with WT hematopoietic cells remained resistant to TNF-induced shock, revealing that a nonhematopoietic lineage mediated protection. TNF-treated Ripk1D138N/D138N mice exhibited no significant increases in intestinal or vascular permeability, nor did they activate the clotting cascade. We show that TNF administration damaged the liver vascular endothelium and induced phosphorylated mixed lineage kinase domain-like (phospho-MLKL) reactivity in endothelial cells isolated from TNF/zVAD-treated WT, but not Ripk1D138N/D138N, mice. These data reveal that the tissue damage present in this SIRS model is reflected, in part, by breaks in the vasculature due to endothelial cell necroptosis and thereby predict that RIPK1 kinase inhibitors may provide clinical benefit to shock and/or sepsis patients.

Authors

Matija Zelic, Justine E. Roderick, Joanne A. O’Donnell, Jesse Lehman, Sung Eun Lim, Harish P. Janardhan, Chinmay M. Trivedi, Manolis Pasparakis, Michelle A. Kelliher

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Figure 5

Nonhematopoietic RIPK1 kinase–inactive cells protect against TNF-induced intestinal and vascular hyperpermeability and coagulation.

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Nonhematopoietic RIPK1 kinase–inactive cells protect against TNF-induced...
(A) FITC-labeled dextran beads were administered to WT and Ripk1D138N/D138N mice by oral gavage 2 hours after TNF injection. Blood was isolated by cardiac puncture and the levels of FITC-dextran in plasma measured by fluorometry (n = 4–7). (B) Bacterial growth in spleen and liver lysates isolated from untreated WT and Ripk1D138N/D138N mice or 10 hours after injection with 15 μg TNF (n = 3–6). (C) WT and Ripk1D138N/D138N mice were injected with TNF and Evans blue dye, organs were collected, and the amount of dye extracted from the lung and liver was measured by spectrophotometry (n = 7–8). (D) Tissue factor levels were determined by ELISA in the plasma of TNF- or TNF/zVAD-treated WT and Ripk1D138N/D138N mice (n = 3–7). (E) WT and Ripk1D138N/D138N reconstituted mice were injected with TNF and Evans blue dye, and the amount of dye extracted from the lung and liver was measured by spectrophotometry (n = 6–7). (F) Tissue factor levels in the plasma of reconstituted WT and Ripk1D138N/D138N mice treated with TNF (n = 3–5). Data are shown as mean ± SEM. *P < 0.05; **P < 0.01, 2-way ANOVA with post hoc Tukey’s test (A and C), 2-tailed Student’s t test (D), or 1-way ANOVA with post hoc Tukey’s test (E and F).