TY - JOUR
T1 - Consequences of cathepsin C inactivation for membrane exposure of proteinase 3, the target antigen in autoimmune vasculitis
AU - Seren, Seda
AU - Rashed Abouzaid, Maha
AU - Eulenberg-Gustavus, Claudia
AU - Hirschfeld, Josefine
AU - Nasr Soliman, Hala
AU - Jerke, Uwe
AU - N'Guessan, Koffi
AU - Dallet-Choisy, Sandrine
AU - Lesner, Adam
AU - Lauritzen, Conni
AU - Schacher, Beate
AU - Eickholz, Peter
AU - Nagy, Nikoletta
AU - Szell, Marta
AU - Croix, Cécile
AU - Viaud-Massuard, Marie-Claude
AU - Al Farraj Aldosari, Abdullah
AU - Ragunatha, Shivanna
AU - Ibrahim Mostafa, Mostafa
AU - Giampieri, Francesca
AU - Battino, Maurizio
AU - Cornillier, Hélène
AU - Lorette, Gérard
AU - Stephan, Jean-Louis
AU - Goizet, Cyril
AU - Pedersen, John
AU - Gauthier, Francis
AU - Jenne, Dieter E.
AU - Marchand-Adam, Sylvain
AU - Chapple, Iain L
AU - Kettritz, Ralph
AU - Korkmaz, Brice
N1 - © 2018 Seren et al.
PY - 2018/8/10
Y1 - 2018/8/10
N2 - Membrane-bound proteinase 3 (PR3m) is the main target antigen of anti-neutrophil cytoplasmic autoantibodies (ANCA) in granulomatosis with polyangiitis, a systemic small-vessel vasculitis. Binding of ANCA to PR3m triggers neutrophil activation with the secretion of enzymatically active PR3 and related neutrophil serine proteases, thereby contributing to vascular damage. PR3 and related proteases are activated from pro-forms by the lysosomal cysteine protease cathepsin C (CatC) during neutrophil maturation. We hypothesized that pharmacological inhibition of CatC provides an effective measure to reduce PR3m and therefore has implications as a novel therapeutic approach in granulomatosis with polyangiitis. We first studied neutrophilic PR3 from 24 patients with Papillon-Lefèvre syndrome (PLS), a genetic form of CatC deficiency. PLS neutrophil lysates showed a largely reduced but still detectable (0.5-4%) PR3 activity when compared with healthy control cells. Despite extremely low levels of cellular PR3, the amount of constitutive PR3m expressed on the surface of quiescent neutrophils and the typical bimodal membrane distribution pattern were similar to what was observed in healthy neutrophils. However, following cell activation, there was no significant increase in the total amount of PR3m on PLS neutrophils, whereas the total amount of PR3m on healthy neutrophils was significantly increased. We then explored the effect of pharmacological CatC inhibition on PR3 stability in normal neutrophils using a potent cell-permeable CatC inhibitor and a CD34+ hematopoietic stem cell model. Human CD34+ hematopoietic stem cells were treated with the inhibitor during neutrophil differentiation over 10 days. We observed strong reductions in PR3m, cellular PR3 protein, and proteolytic PR3 activity, whereas neutrophil differentiation was not compromised.
AB - Membrane-bound proteinase 3 (PR3m) is the main target antigen of anti-neutrophil cytoplasmic autoantibodies (ANCA) in granulomatosis with polyangiitis, a systemic small-vessel vasculitis. Binding of ANCA to PR3m triggers neutrophil activation with the secretion of enzymatically active PR3 and related neutrophil serine proteases, thereby contributing to vascular damage. PR3 and related proteases are activated from pro-forms by the lysosomal cysteine protease cathepsin C (CatC) during neutrophil maturation. We hypothesized that pharmacological inhibition of CatC provides an effective measure to reduce PR3m and therefore has implications as a novel therapeutic approach in granulomatosis with polyangiitis. We first studied neutrophilic PR3 from 24 patients with Papillon-Lefèvre syndrome (PLS), a genetic form of CatC deficiency. PLS neutrophil lysates showed a largely reduced but still detectable (0.5-4%) PR3 activity when compared with healthy control cells. Despite extremely low levels of cellular PR3, the amount of constitutive PR3m expressed on the surface of quiescent neutrophils and the typical bimodal membrane distribution pattern were similar to what was observed in healthy neutrophils. However, following cell activation, there was no significant increase in the total amount of PR3m on PLS neutrophils, whereas the total amount of PR3m on healthy neutrophils was significantly increased. We then explored the effect of pharmacological CatC inhibition on PR3 stability in normal neutrophils using a potent cell-permeable CatC inhibitor and a CD34+ hematopoietic stem cell model. Human CD34+ hematopoietic stem cells were treated with the inhibitor during neutrophil differentiation over 10 days. We observed strong reductions in PR3m, cellular PR3 protein, and proteolytic PR3 activity, whereas neutrophil differentiation was not compromised.
U2 - 10.1074/jbc.RA118.001922
DO - 10.1074/jbc.RA118.001922
M3 - Article
C2 - 29925593
SN - 0021-9258
VL - 293
SP - 12415
EP - 12428
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 32
ER -