A Nonpeptidic Cathepsin S Activity-Based Probe for Noninvasive Optical Imaging of Tumor-Associated Macrophages

Verdoes, Martijn; Edgington, Laura; Scheeren, Ferenc A.; Leyva, Melissa J.; Blum, Galia; Weiskopf, Kipp; Bachmann, Michael; Ellman, Jonathan A.; Bogyo, Matthew

doi:10.1016/j.chembiol.2012.03.012

Cited by 103 publications

(94 citation statements)

References 31 publications

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“…When we analyzed the labeling pattern by in-gel fluorescence, we observed that only CatS retained activity upon CQ treatment, consistent with prior reports that CatS is capable of retaining activity at neutral pH (V. Turk et al, 2012; Verdoes et al, 2012) (Figure 4D). We also analyzed these samples by western blot to confirm loading (Figure 4D) and cathepsin protease expression.…”

Section: Resultssupporting

confidence: 89%

Bifunctional Probes of Cathepsin Protease Activity and pH Reveal Alterations in Endolysosomal pH during Bacterial Infection

Sanman

Linden

Verdoes

et al. 2016

Cell Chemical Biology

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Summary Cysteine cathepsins are lysosomal proteases involved in regulation of both normal cellular processes and disease. Biochemical studies with peptide substrates indicate that cathepsins have optimal activity at acidic pH and highly attenuated activity at neutral pH. In contrast, there is mounting evidence that cathepsins have biological roles in environments that have non-acidic pH. To further define the specific pH environments where cathepsins act, we designed bifunctional activity-based probes (ABPs) that allow simultaneous analysis of cathepsin protease activity and pH. We use these probes to analyze the steady-state environment of cathepsin activity in macrophages and to measure dynamic changes in activity and pH upon stimulation. We show that Salmonella typhimurium induces a change in lysosomal pH that ultimately impairs cathepsin activity in both infected cells and a fraction of bystander cells, highlighting a mechanism by which Salmonella can simultaneously flourish within host cells and alter the behavior of nearby uninfected cells.

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Section: Resultssupporting

confidence: 89%

Bifunctional Probes of Cathepsin Protease Activity and pH Reveal Alterations in Endolysosomal pH during Bacterial Infection

Sanman

Linden

Verdoes

et al. 2016

Cell Chemical Biology

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“…Indeed, when we compared the labeling profiles of the prior probe BMV083 to the new BMV117 probe in live RAW 264.7 cells (mouse leukemic monocyte macrophage cell line) we found that BMV117 showed exclusive labeling of cathepsin S, whereas BMV083 also labeled a substantial amount of cathepsin B and, at higher concentrations, cathepsin L (Figure 1b and Figure S1A). 8 However, the gain in selectivity by introducing increased steric bulk in the P2 position came at the cost of a 10-fold decrease in potency towards cathepsin S. We recently reported the optimized near-infrared pan-reactive cysteine cathepsin qABP BMV109 that contains a more reactive phenoxymethyl ketone (PMK) electrophile (also known as the “warhead”), a shorter spacer and a more hydrophilic sulfo-QSY21 quencher. This probe targets a broad panel of cysteine cathepsin targets and has better aqueous solubility and improved in vivo properties compared to the previous generation acyloxymethyl ketone (AOMK) probes.…”

Section: Resultsmentioning

confidence: 99%

Design of a Highly Selective Quenched Activity-Based Probe and Its Application in Dual Color Imaging Studies of Cathepsin S Activity Localization

Bender¹,

Ofori²,

Linden³

et al. 2015

J. Am. Chem. Soc.

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The cysteine cathepsins are a group of 11 proteases whose function was originally believed to be the degradation of endocytosed material with a high degree of redundancy. However, it has become clear that these enzymes are also important regulators of both health and disease. Thus, selective tools that can discriminate between members of this highly related class of enzymes will be critical to further delineate the unique biological functions of individual cathepsins. Here we present the design and synthesis of a near-infrared quenched activity-based probe (qABP) that selectively targets cathepsin S which is highly expressed in immune cells. Importantly, this high degree of selectivity is retained both in vitro and in vivo. In combination with a new green-fluorescent pan-reactive cysteine cathepsin qABP we performed dual color labeling studies in bone marrow derived immune cells and identified vesicles containing exclusively cathepsin S activity. This observation demonstrates the value of our complimentary cathepsin probes and provides evidence for the existence of specific localization of cathepsin S activity in dendritic cells.

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“…Eight hours after treatment, mice were euthanized and perfused with PBS, and samples of heart, liver, lung, spleen, pancreas, and kidney were collected and frozen in liquid nitrogen. Organs were lysed in citrate buffer, and cathepsin activity was measured as previously described (34,35). Briefly, 50 μg of protein lysate was labeled for 1 h at room temperature with a mixture of 1 μM FY01 and 1 μM DCG04, and run on an SDS/PAGE gel.…”

Section: Methodsmentioning

confidence: 99%

Ferrous iron-dependent drug delivery enables controlled and selective release of therapeutic agents in vivo

Deu¹,

Chen

Lauterwasser

et al. 2013

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

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The precise targeting of cytotoxic agents to specific cell types or cellular compartments is of significant interest in medicine, with particular relevance for infectious diseases and cancer. Here, we describe a method to exploit aberrant levels of mobile ferrous iron (Fe II ) for selective drug delivery in vivo. This approach makes use of a 1,2,4-trioxolane moiety, which serves as an Fe II -sensitive "trigger," making drug release contingent on Fe II -promoted trioxolane fragmentation. We demonstrate in vivo validation of this approach with the Plasmodium berghei model of murine malaria. Malaria parasites produce high concentrations of mobile ferrous iron as a consequence of their catabolism of host hemoglobin in the infected erythrocyte. Using activity-based probes, we successfully demonstrate the Fe II -dependent and parasite-selective delivery of a potent dipeptidyl aminopeptidase inhibitor. We find that delivery of the compound in its Fe II -targeted form leads to more sustained target inhibition with greatly reduced off-target inhibition of mammalian cathepsins. This selective drug delivery translates into improved efficacy and tolerability. These findings demonstrate the utility of a purely chemical means to achieve selective drug targeting in vivo. This approach may find useful application in parasitic infections and more broadly in any disease state characterized by aberrant production of reactive ferrous iron.iron-mediated delivery | targeted prodrugs | dipeptidyl peptidase | combination therapy

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A Nonpeptidic Cathepsin S Activity-Based Probe for Noninvasive Optical Imaging of Tumor-Associated Macrophages

Cited by 103 publications

References 31 publications

Bifunctional Probes of Cathepsin Protease Activity and pH Reveal Alterations in Endolysosomal pH during Bacterial Infection

Bifunctional Probes of Cathepsin Protease Activity and pH Reveal Alterations in Endolysosomal pH during Bacterial Infection

Design of a Highly Selective Quenched Activity-Based Probe and Its Application in Dual Color Imaging Studies of Cathepsin S Activity Localization

Ferrous iron-dependent drug delivery enables controlled and selective release of therapeutic agents in vivo

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