title: Distinct cathepsins control necrotic cell death mediated by pyroptosis inducers and lysosome-destabilizing agents.
abstract: Necrotic cell death triggers a range of biological responses including a strong adaptive immune response, yet we know little about the cellular pathways that control necrotic cell death. Inhibitor studies suggest that proteases, and in particular cathepsins, drive necrotic cell death. The cathepsin B-selective inhibitor CA-074-Me blocks all forms of programmed necrosis by an unknown mechanism. We found that cathepsin B deficiency does not prevent induction of pyroptosis and lysosome-mediated necrosis suggesting that CA-074-Me blocks necrotic cell death by targeting cathepsins other than cathepsin B. A single cathepsin, cathepsin C, drives necrotic cell death mediated by the lysosome-destabilizing agent Leu-Leu-OMe (LLOMe). Here we present evidence that cathepsin C-deficiency and CA-074-Me block LLOMe killing in a distinct and cell type-specific fashion. Cathepsin C-deficiency and CA-074-Me block LLOMe killing of all myeloid cells, except for neutrophils. Cathepsin C-deficiency, but not CA-074-Me, blocks LLOMe killing of neutrophils suggesting that CA-074-Me does not target cathepsin C directly, consistent with inhibitor studies using recombinant cathepsin C. Unlike other cathepsins, cathepsin C lacks endoproteolytic activity, and requires activation by other lysosomal proteases, such as cathepsin D. Consistent with this theory, we found that lysosomotropic agents and cathepsin D downregulation by siRNA block LLOMe-mediated necrosis. Our findings indicate that a proteolytic cascade, involving cathepsins C and D, controls LLOMe-mediated necrosis. In contrast, cathepsins C and D were not required for pyroptotic cell death suggesting that distinct cathepsins control pyroptosis and lysosome-mediated necrosis.
authors.affiliation: a Department of Microbiology and Immunology; Albert Einstein College of Medicine , Bronx , NY USA.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Brojatsch
authors.forename: Heriberto
authors.initials: H
authors.lastname: Lima
authors.forename: Deborah
authors.initials: D
authors.lastname: Palliser
authors.forename: Lee S
authors.initials: LS
authors.lastname: Jacobson
authors.forename: Stefan M
authors.initials: SM
authors.lastname: Muehlbauer
authors.forename: Raquel
authors.initials: R
authors.lastname: Furtado
authors.forename: David L
authors.initials: DL
authors.lastname: Goldman
authors.forename: Michael P
authors.initials: MP
authors.lastname: Lisanti
authors.forename: Kartik
authors.initials: K
authors.lastname: Chandran
mesh_terms: D000818:Animals; D017209:Apoptosis; D002401:Cathepsin B / Q000037:antagonists & inhibitors / Q000502:physiology; D020789:Cathepsin C / Q000502:physiology*; D002402:Cathepsin D / Q000502:physiology*; D004151:Dipeptides / Q000494:pharmacology; D008247:Lysosomes / Q000187:drug effects / Q000201:enzymology*; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D009336:Necrosis
chemical_list: C400541:CA 074 methyl ester; D004151:Dipeptides; C045139:leucyl-leucine-methyl ester; D020789:Cathepsin C; D002401:Cathepsin B; D002402:Cathepsin D
keywords: Caspase-1; LLOMe, L-leucyl-L-leucine methyl ester; LMN, lysosome-mediated necrosis, IL, interleukin; LPS, lipopolysaccharide; NLR, nucleotide-binding domain and leucine rich repeat; inflammasome; lysosome rupture; necrosis; pyroptosis
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