TY - JOUR
T1 - Cathepsin S is activated during colitis and causes visceral hyperalgesia by a PAR2-dependent mechanism in mice
AU - Cattaruzza, Fiore
AU - Lyo, Victoria
AU - Jones, Ella
AU - Pham, David
AU - Hawkins, James
AU - Kirkwood, Kimberley
AU - Valdez-Morales, Eduardo
AU - Ibeakanma, Charles
AU - Vanner, Stephen J.
AU - Bogyo, Matthew
AU - Bunnett, Nigel W.
N1 - Funding Information:
Funding Supported by National Institutes of Health (NIH) grants DK43207 , DK57850 , DK39957 (NWB), Crohn's and Colitis Foundation of Canada (SJV), and NIH grant EB005011 (MB).
PY - 2011/11
Y1 - 2011/11
N2 - Background & Aims: Although proteases control inflammation and pain, the identity, cellular origin, mechanism of action, and causative role of proteases that are activated during disease are not defined. We investigated the activation and function of cysteine cathepsins (Cat) in colitis. Methods: Because protease activity, rather than expression, is regulated, we treated mice with fluorescent activity-based probes that covalently modify activated cathepsins. Activated proteases were localized by tomographic imaging of intact mice and confocal imaging of tissues, and were identified by electrophoresis and immunoprecipitation. We examined the effects of activated cathepsins on excitability of colonic nociceptors and on colonic pain, and determined their role in colonic inflammatory pain by gene deletion. Results: Tomography and magnetic resonance imaging localized activated cathepsins to the inflamed colon of piroxicam-treated il10-/- mice. Confocal imaging detected activated cathepsins in colonic macrophages and spinal neurons and microglial cells of mice with colitis. Gel electrophoresis and immunoprecipitation identified activated Cat-B, Cat-L, and Cat-S in colon and spinal cord, and Cat-S was preferentially secreted into the colonic lumen. Intraluminal Cat-S amplified visceromotor responses to colorectal distension and induced hyperexcitability of colonic nociceptors, which required expression of protease-activated receptor-2. Cat-S deletion attenuated colonic inflammatory pain induced with trinitrobenzene sulfonic acid. Conclusions: Activity-based probes enable noninvasive detection, cellular localization, and proteomic identification of proteases activated during colitis and are potential diagnostic tools for detection of predictive disease biomarkers. Macrophage cathepsins are activated during colitis, and Cat-S activates nociceptors to induce visceral pain via protease-activated receptor-2. Cat-S mediates colitis pain and is a potential therapeutic target.
AB - Background & Aims: Although proteases control inflammation and pain, the identity, cellular origin, mechanism of action, and causative role of proteases that are activated during disease are not defined. We investigated the activation and function of cysteine cathepsins (Cat) in colitis. Methods: Because protease activity, rather than expression, is regulated, we treated mice with fluorescent activity-based probes that covalently modify activated cathepsins. Activated proteases were localized by tomographic imaging of intact mice and confocal imaging of tissues, and were identified by electrophoresis and immunoprecipitation. We examined the effects of activated cathepsins on excitability of colonic nociceptors and on colonic pain, and determined their role in colonic inflammatory pain by gene deletion. Results: Tomography and magnetic resonance imaging localized activated cathepsins to the inflamed colon of piroxicam-treated il10-/- mice. Confocal imaging detected activated cathepsins in colonic macrophages and spinal neurons and microglial cells of mice with colitis. Gel electrophoresis and immunoprecipitation identified activated Cat-B, Cat-L, and Cat-S in colon and spinal cord, and Cat-S was preferentially secreted into the colonic lumen. Intraluminal Cat-S amplified visceromotor responses to colorectal distension and induced hyperexcitability of colonic nociceptors, which required expression of protease-activated receptor-2. Cat-S deletion attenuated colonic inflammatory pain induced with trinitrobenzene sulfonic acid. Conclusions: Activity-based probes enable noninvasive detection, cellular localization, and proteomic identification of proteases activated during colitis and are potential diagnostic tools for detection of predictive disease biomarkers. Macrophage cathepsins are activated during colitis, and Cat-S activates nociceptors to induce visceral pain via protease-activated receptor-2. Cat-S mediates colitis pain and is a potential therapeutic target.
KW - Activity-Based Probes
KW - Inflammation
KW - Pain
KW - Protease-Activated Receptors
KW - Proteases
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U2 - 10.1053/j.gastro.2011.07.035
DO - 10.1053/j.gastro.2011.07.035
M3 - Article
C2 - 21802389
AN - SCOPUS:80054846758
SN - 0016-5085
VL - 141
SP - 1864-1874.e3
JO - Gastroenterology
JF - Gastroenterology
IS - 5
ER -