Protease-Activated Receptor 2, Dipeptidyl Peptidase I, and Proteases Mediate Clostridium difficile Toxin A Enteritis

Cottrell, Graeme S.; Amadesi, Silvia; Pikios, Stella; Camerer, Eric; Willardsen, J. Adam; Murphy, Brett R.; Caughey, George H.; Wolters, Paul J.; Coughlin, Shaun R.; Peterson, Anders; Knecht, Wolfgang; Pothoulakis, Charalabos; Bunnett, Nigel W.; Grady, Eileen F.

doi:10.1053/j.gastro.2007.03.101

Cited by 49 publications

(40 citation statements)

References 56 publications

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“…We previously found that loss of mucin in goblet cells accompanied TxA-mediated epithelial damage (17). Because epithelial architecture was preserved in rats pretreated with CLR dsRNA before TxA treatment (Fig.…”

Section: Resultsmentioning

confidence: 81%

“…An early event in epithelial damage induced by TxA is the loss of mucin from goblet cells (2), which did not occur after pretreatment with CLR dsRNA. We have found that in TxA ileitis, protease-activated receptor 2 −/− mice have an improved outcome and a minimal loss of mucin (17). CGRP can recruit mast cells to sites of inflammation (28) and, in response to TxA, mast cells can alter microvascular tone and promote fluid secretion (2).…”

Section: Discussionmentioning

confidence: 99%

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Local injection of dsRNA targeting calcitonin receptor-like receptor (CLR) ameliorates Clostridium difficile toxin A-induced ileitis

Bhargava¹,

Clifton²,

Mhaske³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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Enteritis caused by Clostridium difficile toxin (Tx) is a nosocomial disease of increasing clinical concern, but the local mediators of C. difficile TxA inflammation are unknown. The potent vasodilator calcitonin gene-related peptide mediates neurogenic inflammation via the calcitonin receptor-like receptor (CLR). Here we examined the ileum-specific effects of reducing CLR on TxA ileitis by local preinjection of double-stranded RNAs. Treatment with CLR dsRNA for 7 d decreased CLR immunoreactivity, whereas treatment with non-CLR dsRNA did not. Subsequent injection of TxA in the same location increased CLR in rats treated with non-CLR dsRNA but not in rats treated with CLR dsRNA, documenting that local injection of dsRNA is effective in preventing the increase in CLR immunoreactivity in response to local TxA. After non-CLR dsRNA pretreatment, TxA induced robust intestinal secretion, myeloperoxidase activity, and histopathologic indications of inflammation including epithelial damage, congestion, neutrophil infiltration, loss of mucin from goblet cells, and increase in mast cell numbers. After CLR dsRNA pretreatment, TxA-induced changes in intestinal secretion and histopathologic inflammation were improved, including normal mucin staining and fewer resident mast cells. Loss of CLR prevented TxA-mediated activation of NF-κB and concomitant increases in pERK1/2 and TNF-α mRNA. Locally produced CLR plays a proinflammatory role in TxA ileitis via MAPK signaling and TNF-α. The results reported here strongly suggest that a local injection of dsRNA targeting CLR could be an effective local therapeutic approach at the inflammation site in the treatment of a growing, clinically relevant hospital-acquired disease, C. difficile infection.

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Section: Resultsmentioning

confidence: 81%

Section: Discussionmentioning

confidence: 99%

Local injection of dsRNA targeting calcitonin receptor-like receptor (CLR) ameliorates Clostridium difficile toxin A-induced ileitis

Bhargava¹,

Clifton²,

Mhaske³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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“…Given that trypsinogen IV lacks a discernable signal peptide, it remains to be determined whether the trypsinogen IV is secreted, although the detection of immunoreactive trypsinogen IV in the extracellular matrix of human brain suggests secretion (12), but by mechanisms that remain to be determined. Moreover, trypsinogen IV is co-expressed with enteropeptidase in many epithelial cells and tissues (10), and inflammatory mediators can up-regulate trypsinogen IV expression in epithelial cells (58). In addition.…”

Section: Discussionmentioning

confidence: 99%

Trypsin IV or Mesotrypsin and p23 Cleave Protease-activated Receptors 1 and 2 to Induce Inflammation and Hyperalgesia

Knecht

Cottrell

Amadesi

et al. 2007

Journal of Biological Chemistry

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Although principally produced by the pancreas to degrade dietary proteins in the intestine, trypsins are also expressed in the nervous system and in epithelial tissues, where they have diverse actions that could be mediated by protease-activated receptors (PARs). We examined the biological actions of human trypsin IV (or mesotrypsin) and rat p23, inhibitor-resistant forms of trypsin. The zymogens trypsinogen IV and pro-p23 were expressed in Escherichia coli and purified to apparent homogeneity. Enteropeptidase cleaved both zymogens, liberating active trypsin IV and p23, which were resistant to soybean trypsin inhibitor and aprotinin. (trypsin IV and p23) mice. Trypsin IV and p23 caused thermal hyperalgesia and mechanical allodynia and hyperalgesia in mice, and these effects were absent in PAR 2 ؊/؊ mice but maintained in PAR 1 ؊/؊ mice. Thus, trypsin IV and p23 are inhibitor-resistant trypsins that can cleave and activate PARs, causing PAR 1 -and PAR 2 -dependent inflammation and PAR 2 -dependent hyperalgesia.

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“…Furthermore, intracolonic PAR2 activation (using trypsin, tryptase, or the peptide SLIGRL-NH 2 ) results in mucosal damage, bowel wall thickening, and myeloperoxidase (MPO) activity, which do not occur in PAR2(Ϫ/Ϫ) mice (Cenac et al, 2002;Patel and Shah, 2010). PAR2(Ϫ/Ϫ) mice also do not develop most symptoms of 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced (Hyun et al, 2008) or bacteria-induced (Hansen et al, 2005;Cottrell et al, 2007) colitis. An efficacious PAR2 antagonist could help to clarify in vivo roles for PAR2 and its activating proteases in the etiology of IBD and validate a new therapeutic avenue for treating IBD.…”

Section: Introductionmentioning

confidence: 99%

Antagonism of Protease-Activated Receptor 2 Protects against Experimental Colitis

Lohman

Cotterell²,

Suen³

et al. 2011

J Pharmacol Exp Ther

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Many trypsin-like serine proteases such as ␤-tryptase are involved in the pathogenesis of colitis and inflammatory bowel diseases. Inhibitors of individual proteases show limited efficacy in treating such conditions, but also probably disrupt digestive and defensive functions of proteases. Here, we investigate whether masking their common target, protease-activated receptor 2 (PAR2), is an effective therapeutic strategy for treating acute and chronic experimental colitis in rats. A novel PAR2 antagonist (5-isoxazoyl-Cha-Ile-spiro[indene-1,4Ј-piperidine]; GB88) was evaluated for the blockade of intracellular calcium release in colonocytes and anti-inflammatory activity in acute (PAR2 agonist-induced) versus chronic [2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced] models of colitis in Wistar rats. Disease progression (disease activity index, weight loss, and mortality) and postmortem colonic histopathology (inflammation, bowel wall thickness, and myeloperoxidase) were measured. PAR2 and tryptase colocalization were investigated by using immunohistochemistry. GB88 was a more potent antagonist of PAR2 activation in colonocytes than another reported compound, N 1 -3-methylbutyryl-N 4 -6-aminohexanoyl-piperazine (ENMD-1068) (IC 50 8 M versus 5 mM). Acute colonic inflammation induced in rats by the PAR2 agonist SLIGRL-NH 2 was inhibited by oral administration of GB88 (10 mg/kg) with markedly reduced edema, mucin depletion, PAR2 receptor internalization, and mastocytosis. Chronic TNBS-induced colitis in rats was ameliorated by GB88 (10 mg/kg/day p.o.), which reduced mortality and pathology (including colon obstruction, ulceration, wall thickness, and myeloperoxidase release) more effectively than the clinically used drug sulfasalazine (100 mg/ kg/day p.o.). These disease-modifying properties for the PAR2 antagonist in both acute and chronic experimental colitis strongly support a pathogenic role for PAR2 and PAR2-activating proteases and therapeutic potential for PAR2 antagonism in inflammatory diseases of the colon.

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Protease-Activated Receptor 2, Dipeptidyl Peptidase I, and Proteases Mediate Clostridium difficile Toxin A Enteritis

Abstract: Background & Aims-We studied the role of protease-activated receptor 2 (PAR 2 ) and its activating enzymes, trypsins and tryptase, in Clostridium difficile toxin A (TxA)-induced enteritis.

Cited by 49 publications

References 56 publications

Local injection of dsRNA targeting calcitonin receptor-like receptor (CLR) ameliorates Clostridium difficile toxin A-induced ileitis

Local injection of dsRNA targeting calcitonin receptor-like receptor (CLR) ameliorates Clostridium difficile toxin A-induced ileitis

Trypsin IV or Mesotrypsin and p23 Cleave Protease-activated Receptors 1 and 2 to Induce Inflammation and Hyperalgesia

Antagonism of Protease-Activated Receptor 2 Protects against Experimental Colitis

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