Our recent discovery that specific enzymes can be detected by near infrared optical imaging in vivo was fortuitous and has avalanched a series of research projects to extend preliminary observations The long term goal of this research is to directly image tumoral protease activity (e.g. lysosomal enzymes such as cathepsins or secreted enzymes such as matrix metalloproteinases) in vivo. Endogenous tumor proteases have been implicated in angiogenesis, local aggression and metastases formation and have received attention as therapeutic targets. Exogenous proteases (e.g. from viruses) are also targets for therapeutic intervention and can potentially be used to image gene expression (see Project 3). In the current project we hypothesize that tumor associated proteases, for example cathepsin D, can be used as unique molecular targets for diagnostic purposes to 1) to study protease activity during tumorigenesis, regression and relapse, 2) facilitate molecular characterization of tumors (e.g. metastatic potential) and 3) improve tumor detection. In prior research we have developed first generation auto-quenched near infrared fluorescent (NIRF) probes that become detectable after protease activation and have shown that this quenching/dequenching strategy represents a unique amplification strategy increasing target/background ratio 10-100 fold (Weissleder et al., Nature Biotech 199:375-378). Cell culture, in vitro and in vivo studies confirmed that the probes had very low fluorescence unless activated by proteases and were detectable in nanomole amounts in vivo with no apparent toxicity at concentrations tested. In subsequent studies we have generated second generation probes with enzyme specificity (e.g. cathepsin D), higher fluorochrome payload and better quenching/dequenching characteristics. Using a cathepsin D negative and cathepsin D positive mouse tumor models we have shown the specificity of this approach. The proposed studies are a logical extension of preliminary feasibility studies and will apply the developed technique to imaging of cathepsin D known to be over-expressed 2-50 fold in breast cancers. Overall, we expect that this novel technique will potentially allow to image specific enzymes in living organism which would have important applications in tumor detection and therapy assessment.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Specialized Center (P50)
Project #
1P50CA086355-01
Application #
6401938
Study Section
Special Emphasis Panel (ZCA1)
Project Start
2000-08-09
Project End
2005-07-31
Budget Start
Budget End
Support Year
1
Fiscal Year
2000
Total Cost
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02199
Dubach, J Matthew; Kim, Eunha; Yang, Katherine et al. (2017) Quantitating drug-target engagement in single cells in vitro and in vivo. Nat Chem Biol 13:168-173
Vinegoni, Claudio; Fumene Feruglio, Paolo; Brand, Christian et al. (2017) Measurement of drug-target engagement in live cells by two-photon fluorescence anisotropy imaging. Nat Protoc 12:1472-1497
Iaconelli, Jonathan; Lalonde, Jasmin; Watmuff, Bradley et al. (2017) Lysine Deacetylation by HDAC6 Regulates the Kinase Activity of AKT in Human Neural Progenitor Cells. ACS Chem Biol 12:2139-2148
Arlauckas, Sean P; Garris, Christopher S; Kohler, Rainer H et al. (2017) In vivo imaging reveals a tumor-associated macrophage-mediated resistance pathway in anti-PD-1 therapy. Sci Transl Med 9:
Miller, Miles A; Weissleder, Ralph (2017) Imaging the pharmacology of nanomaterials by intravital microscopy: Toward understanding their biological behavior. Adv Drug Deliv Rev 113:61-86
Engblom, Camilla; Pfirschke, Christina; Zilionis, Rapolas et al. (2017) Osteoblasts remotely supply lung tumors with cancer-promoting SiglecFhigh neutrophils. Science 358:
Miller, Miles A; Askevold, Bjorn; Mikula, Hannes et al. (2017) Nano-palladium is a cellular catalyst for in vivo chemistry. Nat Commun 8:15906
Pucci, Ferdinando; Garris, Christopher; Lai, Charles P et al. (2016) SCS macrophages suppress melanoma by restricting tumor-derived vesicle-B cell interactions. Science 352:242-6
Roy, Jeremy; Kim, Bongki; Hill, Eric et al. (2016) Tyrosine kinase-mediated axial motility of basal cells revealed by intravital imaging. Nat Commun 7:10666
Pfirschke, Christina; Engblom, Camilla; Rickelt, Steffen et al. (2016) Immunogenic Chemotherapy Sensitizes Tumors to Checkpoint Blockade Therapy. Immunity 44:343-54

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