Cytotoxicity of crystals involves RIPK3-MLKL-mediated necroptosis

Nat Commun. 2016 Jan 28;7:10274. doi: 10.1038/ncomms10274.

Abstract

Crystals cause injury in numerous disorders, and induce inflammation via the NLRP3 inflammasome, however, it remains unclear how crystals induce cell death. Here we report that crystals of calcium oxalate, monosodium urate, calcium pyrophosphate dihydrate and cystine trigger caspase-independent cell death in five different cell types, which is blocked by necrostatin-1. RNA interference for receptor-interacting protein kinase 3 (RIPK3) or mixed lineage kinase domain like (MLKL), two core proteins of the necroptosis pathway, blocks crystal cytotoxicity. Consistent with this, deficiency of RIPK3 or MLKL prevents oxalate crystal-induced acute kidney injury. The related tissue inflammation drives TNF-α-related necroptosis. Also in human oxalate crystal-related acute kidney injury, dying tubular cells stain positive for phosphorylated MLKL. Furthermore, necrostatin-1 and necrosulfonamide, an inhibitor for human MLKL suppress crystal-induced cell death in human renal progenitor cells. Together, TNF-α/TNFR1, RIPK1, RIPK3 and MLKL are molecular targets to limit crystal-induced cytotoxicity, tissue injury and organ failure.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis*
  • Calcium Oxalate / chemistry
  • Calcium Oxalate / metabolism
  • Calcium Oxalate / toxicity*
  • Calcium Pyrophosphate / chemistry
  • Calcium Pyrophosphate / metabolism
  • Calcium Pyrophosphate / toxicity*
  • Humans
  • Kidney Diseases / genetics
  • Kidney Diseases / metabolism
  • Kidney Diseases / physiopathology*
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Necrosis
  • Phosphorylation
  • Protein Kinases / genetics
  • Protein Kinases / metabolism*
  • Receptor-Interacting Protein Serine-Threonine Kinases / genetics
  • Receptor-Interacting Protein Serine-Threonine Kinases / metabolism*
  • Tumor Necrosis Factor-alpha / genetics
  • Tumor Necrosis Factor-alpha / metabolism
  • Uric Acid / chemistry
  • Uric Acid / metabolism
  • Uric Acid / toxicity*

Substances

  • Tumor Necrosis Factor-alpha
  • Calcium Oxalate
  • Uric Acid
  • MLKL protein, mouse
  • Protein Kinases
  • Receptor-Interacting Protein Serine-Threonine Kinases
  • Ripk3 protein, mouse
  • Calcium Pyrophosphate