While we now have relatively complete lists of all proteases expressed in any given organism, our understanding of the functional roles of these enzymes, for the most part, remains largely unclear. The development of new technologies that will allow the imaging of protease activity is a critical step to begin to map out complex proteolytic cascades as well as to validate proteases as biomarkers for human disease. In the first phase of funding for this project we developed fluorescently labeled small molecule activity based probes (ABPs) that target two major classes of cystiene proteases, the cathepsins and caspases. We then demonstrated that fluorescent ABPs could be used for functional, non-invasive imaging studies of these proteases in various mouse models of human cancer. In the next phase of the project we will apply the probes from the initial funding period to mouse models of inflammation and atherosclerosis. We will also engineer several new classes of imaging probes that target proteases that play roles in the regulation of inflammation. These targets include cathepsins S and K, caspases-1, -4 and -5 and the immunoproteasome. We believe that these reagents will expand the use of the ABP technology platform to a large number of important human diseases that involve the process of inflammation.

Public Health Relevance

This project outlines plans to develop fluorescently labeled imaging agents that target proteases involved in the process of inflammation. Specifically these reagents will be used to detect inflammation in atherosclerotic plaques and may ultimately lead to the generation of contrast agents for early diagnosis and disease monitoring using non-invasive imaging methods.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
5R01EB005011-07
Application #
8204858
Study Section
Microscopic Imaging Study Section (MI)
Program Officer
Conroy, Richard
Project Start
2005-04-01
Project End
2013-12-31
Budget Start
2012-01-01
Budget End
2012-12-31
Support Year
7
Fiscal Year
2012
Total Cost
$458,267
Indirect Cost
$173,747
Name
Stanford University
Department
Pathology
Type
Schools of Medicine
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94305
Li, Hao; Bogyo, Matthew; da Fonseca, Paula C A (2016) The cryo-EM structure of the Plasmodium falciparum 20S proteasome and its use in the fight against malaria. FEBS J 283:4238-4243
Garland, Megan; Yim, Joshua J; Bogyo, Matthew (2016) A Bright Future for Precision Medicine: Advances in Fluorescent Chemical Probe Design and Their Clinical Application. Cell Chem Biol 23:122-36
Lentz, Christian S; Ordonez, Alvaro A; Kasperkiewicz, Paulina et al. (2016) Design of Selective Substrates and Activity-Based Probes for Hydrolase Important for Pathogenesis 1 (HIP1) from Mycobacterium tuberculosis. ACS Infect Dis :
van der Linden, Wouter A; Schulze, Christopher J; Herbert, Andrew S et al. (2016) Cysteine Cathepsin Inhibitors as Anti-Ebola Agents. ACS Infect Dis 2:173-179
Sanman, Laura E; Qian, Yu; Eisele, Nicholas A et al. (2016) Disruption of glycolytic flux is a signal for inflammasome signaling and pyroptotic cell death. Elife 5:e13663
Li, Hao; O'Donoghue, Anthony J; van der Linden, Wouter A et al. (2016) Structure- and function-based design of Plasmodium-selective proteasome inhibitors. Nature 530:233-6
Withana, Nimali P; Ma, Xiaowei; McGuire, Helen M et al. (2016) Non-invasive Imaging of Idiopathic Pulmonary Fibrosis Using Cathepsin Protease Probes. Sci Rep 6:19755
Withana, Nimali P; Garland, Megan; Verdoes, Martijn et al. (2016) Labeling of active proteases in fresh-frozen tissues by topical application of quenched activity-based probes. Nat Protoc 11:184-91
Sanman, Laura E; van der Linden, Wouter A; Verdoes, Martijn et al. (2016) Bifunctional Probes of Cathepsin Protease Activity and pH Reveal Alterations in Endolysosomal pH during Bacterial Infection. Cell Chem Biol 23:793-804
Segal, Ehud; Prestwood, Tyler R; van der Linden, Wouter A et al. (2015) Detection of intestinal cancer by local, topical application of a quenched fluorescence probe for cysteine cathepsins. Chem Biol 22:148-58

Showing the most recent 10 out of 93 publications