An Essential Role for SHARPIN in the Regulation of Caspase 1 Activity in Sepsis

Am J Pathol. 2016 May;186(5):1206-20. doi: 10.1016/j.ajpath.2015.12.026. Epub 2016 Mar 8.

Abstract

Sepsis is burdened by high mortality due to uncontrolled inflammatory response to pathogens. Increased caspase 1 activation causing maturation of IL1β/18 remains a therapeutic challenge in sepsis. SHARPIN (shank-associated regulator of G-protein signaling homology domain-interacting protein), a component of the LUBAC (linear ubiquitin chain-assembly complex), regulates inflammation, with unknown effects on caspase 1 activation. Mice lacking Casp1, Casp11, or both in a Sharpin-deficient background were generated, exposed to lipopolysaccharide-induced endotoxemia, and injected with caspase 1 inhibitor. We monitored survival, Il1β/18, and caspase 1/11 levels in plasma and organs and deciphered mechanisms of SHARPIN-dependent caspase 1 inhibition. A correlation between LUBAC and active caspase 1 was found in blood mononuclear cells from septic patients. SHARPIN bound caspase 1 and disrupted p20/p10 dimer formation, the last step of caspase 1 processing, thereby inhibiting enzyme activation and maturation of IL1β/18 in a LUBAC-independent manner. In septic patients, LUBAC-independent decline in SHARPIN correlated with enhancement of active caspase 1 in circulating mononuclear cells. Septic Sharpin-deficient mice displayed enrichment in mature Il1β/18 and active caspase 1, and shortened survival. Inhibition of caspase 1 reduced levels of Il1β/18 and splenic cell death, and prolonged survival in septic Sharpin-deficient mice. Our findings identify SHARPIN as a potent in vivo caspase 1 inhibitor and propose the caspase 1-SHARPIN interaction as a target in sepsis.

MeSH terms

  • Animals
  • Caspase 1 / deficiency
  • Caspase 1 / metabolism*
  • Caspase Inhibitors / pharmacology
  • Caspases / deficiency
  • Caspases / metabolism
  • Caspases, Initiator
  • Cells, Cultured
  • Dermatitis / enzymology
  • Down-Regulation / physiology
  • Endotoxemia / chemically induced
  • Gene Knockdown Techniques
  • Interleukin-18 / metabolism
  • Interleukin-1beta / metabolism
  • Leukocytes, Mononuclear / enzymology
  • Lipopolysaccharides / toxicity
  • Lung / enzymology
  • Mice, Inbred C57BL
  • Mice, Knockout
  • NF-kappa B / pharmacology
  • Nerve Tissue Proteins / deficiency
  • Nerve Tissue Proteins / physiology*
  • Phenotype
  • Salmonella
  • Sepsis / enzymology*
  • Transfection

Substances

  • Caspase Inhibitors
  • Interleukin-18
  • Interleukin-1beta
  • Lipopolysaccharides
  • NF-kappa B
  • Nerve Tissue Proteins
  • sharpin
  • Casp4 protein, mouse
  • Caspases
  • Caspases, Initiator
  • Caspase 1