TGF-ß Regulates Cathepsin Activation during Normal and Pathogenic Development

Abstract

Summary: Cysteine cathepsins play roles during development and disease beyond their function in lysosomal protein turnover. Here, we leverage a fluorescent activity-based probe (ABP), BMV109, to track cysteine cathepsins in normal and diseased zebrafish embryos. Using this probe in a model of mucolipidosis II, we show that loss of carbohydrate-dependent lysosomal sorting alters the activity of several cathepsin proteases. The data support a pathogenic mechanism where TGF-ß signals enhance the proteolytic processing of pro-Ctsk by modulating the expression of chondroitin 4-sulfate (C4-S). In MLII, elevated C4-S corresponds with TGF-ß-mediated increases in chst11 expression. Inhibiting chst11 impairs the proteolytic activation of Ctsk and alleviates the MLII phenotypes. These findings uncover a regulatory loop between TGF-ß signaling and Ctsk activation that is altered in the context of lysosomal disease. This work highlights the power of ABPs to identify mechanisms underlying pathogenic development in living animals. : Chondroitin sulfate is a known regulator of cathepsin protease activity. Flanagan-Steet et al. identify a positive feedback mechanism whereby cathepsins secreted from chondrocytes upon loss of lysosomal targeting activate TGF-ß signaling in developing cartilage. This increased signaling, in turn, stimulates chondroitin-4 sulfation and enhances cathepsin activity. Keywords: activity-based profiling, cathepsin proteases, lysosomes, cartilage, zebrafish, mucolipidosis, glycosylation, glycosaminoglycans