skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Novel, dually radiolabeled peptides for simultaneous monitoring of enzymatic activity and protein targets

Abstract

This application investigated a novel imaging approach to develop methods to incorporate multiple radionuclides into a single peptide at chemoselective sites for simultaneous monitoring of cell-bound protein targets as well as specific enzymatic activity, both of which are associated with enhanced tumor growth and metastasis. This imaging construct was synthesized in such a manner so that the PET radionuclide will remain associated with the tumor cells and the SPECT radionuclide was cleaved from the imaging agent. Measurement of the PET agent only will yield information about the tumor marker density while measurement of the amount of co-localization and mismatch of the two radionuclides will yield information about the enzymatic activity. This coincident measuring technique using both PET and SPECT agents allows us to draw correlations involving the interactions of enzymes (cathepsin, serine-protease urokinase (uPA) and matrix metalloproteases) and other cellular proteins which play a role in cancer growth and metastasis. This technique will allow for studies in xenograft or genetic models of cancer in the same animal at the same time, thus eliminating problems that may occur when trying to invoke comparisons across animals or timepoints. By using radionuclide imaging as opposed to other imaging modalities, this technique has themore » potential to be translatable and can exploit the high specific activity probes which can be generated with radiotracers. The proof of principle test of this system investigated simultaneous monitoring of matrix metalloprotease (MMP) activity in the extracellular matrix (ECM) as well as density of integrins on the cell surface, both of which can serve as tumor markers. The outcomes/deliverables of this project were as follows: 1. Peptides were synthesized dually labeled at chemospecific sites with PET and SPECT agents. 2. Stability (intrinsic and to radiolysis) and specific activity of these labeled compounds were determined. 3. The feasibility of using these agents for simultaneous monitoring of MMP-2 enzymatic activity and ²3 integrin density was demonstrated in several in vitro assays Radiotracers can be detected at concentrations up to 1000 fold lower than those labeled with non-radioactive markers (e.g. MRI contrast agents), thus using this technique has the advantage of very high sensitivity to measure these processes in vivo. Hence, the development of an efficient approach to the dual labeling of these molecular probes is embodied within this project, with the end result yielding a molecular imaging probe with the highest specific activity possible. An advantage to this dual labeling approach is the ability to measure two different biochemical processes at the same time, a benefit which is not possible in scans involving protocols utilizing two different radiolabeled agents injected sequentially. Another advantage to this technique is the ability to measure enzymatic activity in the form of substrate cleavage. This can only be achieved with a dually labeled compound as has been demonstrated in the case of FRET1. To our knowledge this is the first instance of a measurement of enzymatic substrate cleavage by a dually labeled PET/SPECT radionuclide imaging agent.« less

Authors:
Publication Date:
Research Org.:
Suzanne Lapi (Washington University)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1057270
Report Number(s):
DOE/SC0002114-F
DOE Contract Number:  
SC0002114
Resource Type:
Technical Report
Country of Publication:
United States
Language:
English
Subject:
38 RADIATION CHEMISTRY, RADIOCHEMISTRY, AND NUCLEAR CHEMISTRY; Radiochemistry, copper-64, iodine-123, Positron Emission Tomography

Citation Formats

Efrem Mebrahtu, Suzanne Lapi. Novel, dually radiolabeled peptides for simultaneous monitoring of enzymatic activity and protein targets. United States: N. p., 2012. Web.
Efrem Mebrahtu, Suzanne Lapi. Novel, dually radiolabeled peptides for simultaneous monitoring of enzymatic activity and protein targets. United States.
Efrem Mebrahtu, Suzanne Lapi. Thu . "Novel, dually radiolabeled peptides for simultaneous monitoring of enzymatic activity and protein targets". United States.
@article{osti_1057270,
title = {Novel, dually radiolabeled peptides for simultaneous monitoring of enzymatic activity and protein targets},
author = {Efrem Mebrahtu, Suzanne Lapi},
abstractNote = {This application investigated a novel imaging approach to develop methods to incorporate multiple radionuclides into a single peptide at chemoselective sites for simultaneous monitoring of cell-bound protein targets as well as specific enzymatic activity, both of which are associated with enhanced tumor growth and metastasis. This imaging construct was synthesized in such a manner so that the PET radionuclide will remain associated with the tumor cells and the SPECT radionuclide was cleaved from the imaging agent. Measurement of the PET agent only will yield information about the tumor marker density while measurement of the amount of co-localization and mismatch of the two radionuclides will yield information about the enzymatic activity. This coincident measuring technique using both PET and SPECT agents allows us to draw correlations involving the interactions of enzymes (cathepsin, serine-protease urokinase (uPA) and matrix metalloproteases) and other cellular proteins which play a role in cancer growth and metastasis. This technique will allow for studies in xenograft or genetic models of cancer in the same animal at the same time, thus eliminating problems that may occur when trying to invoke comparisons across animals or timepoints. By using radionuclide imaging as opposed to other imaging modalities, this technique has the potential to be translatable and can exploit the high specific activity probes which can be generated with radiotracers. The proof of principle test of this system investigated simultaneous monitoring of matrix metalloprotease (MMP) activity in the extracellular matrix (ECM) as well as density of integrins on the cell surface, both of which can serve as tumor markers. The outcomes/deliverables of this project were as follows: 1. Peptides were synthesized dually labeled at chemospecific sites with PET and SPECT agents. 2. Stability (intrinsic and to radiolysis) and specific activity of these labeled compounds were determined. 3. The feasibility of using these agents for simultaneous monitoring of MMP-2 enzymatic activity and ²3 integrin density was demonstrated in several in vitro assays Radiotracers can be detected at concentrations up to 1000 fold lower than those labeled with non-radioactive markers (e.g. MRI contrast agents), thus using this technique has the advantage of very high sensitivity to measure these processes in vivo. Hence, the development of an efficient approach to the dual labeling of these molecular probes is embodied within this project, with the end result yielding a molecular imaging probe with the highest specific activity possible. An advantage to this dual labeling approach is the ability to measure two different biochemical processes at the same time, a benefit which is not possible in scans involving protocols utilizing two different radiolabeled agents injected sequentially. Another advantage to this technique is the ability to measure enzymatic activity in the form of substrate cleavage. This can only be achieved with a dually labeled compound as has been demonstrated in the case of FRET1. To our knowledge this is the first instance of a measurement of enzymatic substrate cleavage by a dually labeled PET/SPECT radionuclide imaging agent.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2012},
month = {12}
}

Technical Report:
Other availability
Please see Document Availability for additional information on obtaining the full-text document. Library patrons may search WorldCat to identify libraries that may hold this item. Keep in mind that many technical reports are not cataloged in WorldCat.

Save / Share: