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  • Calpain-2 Inhibitor Therapy Reduces Murine Colitis and Colitis-associated Cancer.

Calpain-2 Inhibitor Therapy Reduces Murine Colitis and Colitis-associated Cancer.

Inflammatory bowel diseases (2015-06-16)
Aaron H Rose, Zhi Huang, Chrisy Mafnas, Jared H Hara, FuKun W Hoffmann, Ann S Hashimoto, Pietro Bertino, Peter R Hoffmann
ABSTRACT

An important role has emerged for calpain enzymes in regulating inflammation with one isoform, calpain-2, particularly important for macrophage activation. The goal of this study was to determine the therapeutic potential of a synthetic calpain-2 inhibitor, zLLY-CH2F, for colitis and inflammation-associated colorectal cancer. Mice were then subjected to the azoxymethane/dextran sulfate sodium model of colitis and colitis-associated cancer incorporating intervention with daily injections of 0.75 mg/kg calpain-2 inhibitor beginning after the first signs of colitis. Calpain-2 inhibitor treatment alleviated weight loss and bloody diarrhea, and reduced inflammatory infiltration into colon tissues and inflammatory cytokine mRNA. Calpain-2 inhibitor intervention also reduced total colitis-associated cancer tumor volume by up to 70% in vehicle control mice and decreased cancer pathology scores of blinded histological colon tissue analyses. Mechanistic investigations showed that calpain-2 inhibition during macrophage activation reduced inhibitor of kappa beta (IκB) degradation and nuclear factor kappa beta (NFκB) nuclear localization as well as secretion of specific inflammatory cytokines. In addition, calpain-2 inhibitor treatment of CT26.WT mouse and HT-29 human colorectal cancer cells decreased proliferation and reduced IκB degradation and NFκB translocation. Overall, these findings suggest that intervention with a calpain-2 inhibitor may reduce colitis and colitis-associated cancer through a two-hit process of limiting macrophage activation and inhibiting growth of the colorectal cancer cells themselves.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ethylenediaminetetraacetic acid solution, 0.02% in DPBS (0.5 mM), sterile-filtered, BioReagent, suitable for cell culture
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Ethylenediaminetetraacetic acid, 99.995% trace metals basis
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Dextran sulfate sodium salt from Leuconostoc spp., low sulfate content, Mr ~40,000
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Dextran sulfate sodium salt from Leuconostoc spp., mol wt 6,500-10,000
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Ethylenediaminetetraacetic acid, ACS reagent, 99.4-100.6%, powder
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Dextran sulfate sodium salt from Leuconostoc spp., for molecular biology, average Mw >500,000 (dextran starting material), contains 0.5-2% phosphate buffer
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Dextran sulfate sodium salt from Leuconostoc spp., avg mol wt >500,000 (dextran starting material)
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Dextran sulfate sodium salt from Leuconostoc spp., average mol wt >500,000 (dextran starting material), contains 0.5-2.0% phosphate buffer, pH 6-8
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Dextran sulfate sodium salt from Leuconostoc spp., average mol wt 9,000-20,000
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Ethylenediaminetetraacetic acid, purified grade, ≥98.5%, powder
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Ethylenediaminetetraacetic acid, BioUltra, anhydrous, ≥99% (titration)
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Ethylenediaminetetraacetic acid, anhydrous, crystalline, BioReagent, suitable for cell culture
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Azoxymethane, 13.4 M, ≥98%
Sigma-Aldrich
Dextran sulfate sodium salt from Leuconostoc spp., Mr ~500,000