In vivo positron emission tomography imaging of protease activity by generation of a hydrophobic product from a noninhibitory protease substrate

Chih Hung Chuang, Kuo Hsiang Chuang, Hsin Ell Wang, Steve R. Roffler, Jen Taie Shiea, Shey Cherng Tzou, Ta Chun Cheng, Chien Han Kao, Shih Yen Wu, Wei Lung Tseng, Chiu Min Cheng, Ming Feng Hou, Ju Ming Wang, Tian Lu Cheng

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Purpose: To develop an imaging technology for protease activities in patients that could help in prognosis prediction and in design of personalized, protease-based inhibitors and prodrugs for targeted therapy. Experimental Design: Polyethylene glycol (PEG) was covalently attached to the N-terminus of a hydrophilic peptide substrate (GPLGVR) for matrix metalloproteinase (MMP) to increase hydrophilicity. PEG-peptide was then linked to a hydrophobic tetramethylrhodamine (TMR) domain and labeled with 18F to form a PEG-peptide-18F-TMR probe. Specific cleavage of the probe by MMP2 was tested in vitro by matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF). In vivo imaging of MMP2-expressing tumors was evaluated by micro-PET. Results: The hydrophobic TMR fragment (948 Da) was specifically generated by MMP2 enzymes and MMP-expressing HT1080 cells but not control MCF-7 cells. MMP-expressing HT1080 cells and tumors selectively accumulated the hydrolyzed, hydrophobic TMR fragment at sites of protease activity. Importantly, we found that 18F-labeled probe (18F-TMR) preferentially localized in HT1080 tumors but not control MCF-7 tumors as shown by micro-PET. Uptake of the probe in HT1080 tumors was 18.4 ± 1.9-fold greater than in the MCF-7 tumors 30 minutes after injection. These results suggest that the PEG-peptide-18F-TMR probe displays high selectivity for imaging MMP activity. Conclusions: This strategy successfully images MMP expression in vivo and may be extended to other proteases to predict patient prognosis and to design personalized, protease-based inhibitors and prodrug-targeted therapies.

Original languageEnglish
Pages (from-to)238-247
Number of pages10
JournalClinical Cancer Research
Volume18
Issue number1
DOIs
Publication statusPublished - Jan 1 2012
Externally publishedYes

Fingerprint

Positron-Emission Tomography
Matrix Metalloproteinases
Peptide Hydrolases
Neoplasms
Peptides
Prodrugs
Protease Inhibitors
Matrix Metalloproteinase 2
MCF-7 Cells
Hydrophobic and Hydrophilic Interactions
tetramethylrhodamine
Lasers
Research Design
Technology
Injections
Enzymes
Therapeutics

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

In vivo positron emission tomography imaging of protease activity by generation of a hydrophobic product from a noninhibitory protease substrate. / Chuang, Chih Hung; Chuang, Kuo Hsiang; Wang, Hsin Ell; Roffler, Steve R.; Shiea, Jen Taie; Tzou, Shey Cherng; Cheng, Ta Chun; Kao, Chien Han; Wu, Shih Yen; Tseng, Wei Lung; Cheng, Chiu Min; Hou, Ming Feng; Wang, Ju Ming; Cheng, Tian Lu.

In: Clinical Cancer Research, Vol. 18, No. 1, 01.01.2012, p. 238-247.

Research output: Contribution to journalArticle

Chuang, CH, Chuang, KH, Wang, HE, Roffler, SR, Shiea, JT, Tzou, SC, Cheng, TC, Kao, CH, Wu, SY, Tseng, WL, Cheng, CM, Hou, MF, Wang, JM & Cheng, TL 2012, 'In vivo positron emission tomography imaging of protease activity by generation of a hydrophobic product from a noninhibitory protease substrate', Clinical Cancer Research, vol. 18, no. 1, pp. 238-247. https://doi.org/10.1158/1078-0432.CCR-11-0608
Chuang, Chih Hung ; Chuang, Kuo Hsiang ; Wang, Hsin Ell ; Roffler, Steve R. ; Shiea, Jen Taie ; Tzou, Shey Cherng ; Cheng, Ta Chun ; Kao, Chien Han ; Wu, Shih Yen ; Tseng, Wei Lung ; Cheng, Chiu Min ; Hou, Ming Feng ; Wang, Ju Ming ; Cheng, Tian Lu. / In vivo positron emission tomography imaging of protease activity by generation of a hydrophobic product from a noninhibitory protease substrate. In: Clinical Cancer Research. 2012 ; Vol. 18, No. 1. pp. 238-247.
@article{3d5a4aa234b646d98a4a5da5b2ce9dec,
title = "In vivo positron emission tomography imaging of protease activity by generation of a hydrophobic product from a noninhibitory protease substrate",
abstract = "Purpose: To develop an imaging technology for protease activities in patients that could help in prognosis prediction and in design of personalized, protease-based inhibitors and prodrugs for targeted therapy. Experimental Design: Polyethylene glycol (PEG) was covalently attached to the N-terminus of a hydrophilic peptide substrate (GPLGVR) for matrix metalloproteinase (MMP) to increase hydrophilicity. PEG-peptide was then linked to a hydrophobic tetramethylrhodamine (TMR) domain and labeled with 18F to form a PEG-peptide-18F-TMR probe. Specific cleavage of the probe by MMP2 was tested in vitro by matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF). In vivo imaging of MMP2-expressing tumors was evaluated by micro-PET. Results: The hydrophobic TMR fragment (948 Da) was specifically generated by MMP2 enzymes and MMP-expressing HT1080 cells but not control MCF-7 cells. MMP-expressing HT1080 cells and tumors selectively accumulated the hydrolyzed, hydrophobic TMR fragment at sites of protease activity. Importantly, we found that 18F-labeled probe (18F-TMR) preferentially localized in HT1080 tumors but not control MCF-7 tumors as shown by micro-PET. Uptake of the probe in HT1080 tumors was 18.4 ± 1.9-fold greater than in the MCF-7 tumors 30 minutes after injection. These results suggest that the PEG-peptide-18F-TMR probe displays high selectivity for imaging MMP activity. Conclusions: This strategy successfully images MMP expression in vivo and may be extended to other proteases to predict patient prognosis and to design personalized, protease-based inhibitors and prodrug-targeted therapies.",
author = "Chuang, {Chih Hung} and Chuang, {Kuo Hsiang} and Wang, {Hsin Ell} and Roffler, {Steve R.} and Shiea, {Jen Taie} and Tzou, {Shey Cherng} and Cheng, {Ta Chun} and Kao, {Chien Han} and Wu, {Shih Yen} and Tseng, {Wei Lung} and Cheng, {Chiu Min} and Hou, {Ming Feng} and Wang, {Ju Ming} and Cheng, {Tian Lu}",
year = "2012",
month = "1",
day = "1",
doi = "10.1158/1078-0432.CCR-11-0608",
language = "English",
volume = "18",
pages = "238--247",
journal = "Clinical Cancer Research",
issn = "1078-0432",
publisher = "American Association for Cancer Research Inc.",
number = "1",

}

TY - JOUR

T1 - In vivo positron emission tomography imaging of protease activity by generation of a hydrophobic product from a noninhibitory protease substrate

AU - Chuang, Chih Hung

AU - Chuang, Kuo Hsiang

AU - Wang, Hsin Ell

AU - Roffler, Steve R.

AU - Shiea, Jen Taie

AU - Tzou, Shey Cherng

AU - Cheng, Ta Chun

AU - Kao, Chien Han

AU - Wu, Shih Yen

AU - Tseng, Wei Lung

AU - Cheng, Chiu Min

AU - Hou, Ming Feng

AU - Wang, Ju Ming

AU - Cheng, Tian Lu

PY - 2012/1/1

Y1 - 2012/1/1

N2 - Purpose: To develop an imaging technology for protease activities in patients that could help in prognosis prediction and in design of personalized, protease-based inhibitors and prodrugs for targeted therapy. Experimental Design: Polyethylene glycol (PEG) was covalently attached to the N-terminus of a hydrophilic peptide substrate (GPLGVR) for matrix metalloproteinase (MMP) to increase hydrophilicity. PEG-peptide was then linked to a hydrophobic tetramethylrhodamine (TMR) domain and labeled with 18F to form a PEG-peptide-18F-TMR probe. Specific cleavage of the probe by MMP2 was tested in vitro by matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF). In vivo imaging of MMP2-expressing tumors was evaluated by micro-PET. Results: The hydrophobic TMR fragment (948 Da) was specifically generated by MMP2 enzymes and MMP-expressing HT1080 cells but not control MCF-7 cells. MMP-expressing HT1080 cells and tumors selectively accumulated the hydrolyzed, hydrophobic TMR fragment at sites of protease activity. Importantly, we found that 18F-labeled probe (18F-TMR) preferentially localized in HT1080 tumors but not control MCF-7 tumors as shown by micro-PET. Uptake of the probe in HT1080 tumors was 18.4 ± 1.9-fold greater than in the MCF-7 tumors 30 minutes after injection. These results suggest that the PEG-peptide-18F-TMR probe displays high selectivity for imaging MMP activity. Conclusions: This strategy successfully images MMP expression in vivo and may be extended to other proteases to predict patient prognosis and to design personalized, protease-based inhibitors and prodrug-targeted therapies.

AB - Purpose: To develop an imaging technology for protease activities in patients that could help in prognosis prediction and in design of personalized, protease-based inhibitors and prodrugs for targeted therapy. Experimental Design: Polyethylene glycol (PEG) was covalently attached to the N-terminus of a hydrophilic peptide substrate (GPLGVR) for matrix metalloproteinase (MMP) to increase hydrophilicity. PEG-peptide was then linked to a hydrophobic tetramethylrhodamine (TMR) domain and labeled with 18F to form a PEG-peptide-18F-TMR probe. Specific cleavage of the probe by MMP2 was tested in vitro by matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF). In vivo imaging of MMP2-expressing tumors was evaluated by micro-PET. Results: The hydrophobic TMR fragment (948 Da) was specifically generated by MMP2 enzymes and MMP-expressing HT1080 cells but not control MCF-7 cells. MMP-expressing HT1080 cells and tumors selectively accumulated the hydrolyzed, hydrophobic TMR fragment at sites of protease activity. Importantly, we found that 18F-labeled probe (18F-TMR) preferentially localized in HT1080 tumors but not control MCF-7 tumors as shown by micro-PET. Uptake of the probe in HT1080 tumors was 18.4 ± 1.9-fold greater than in the MCF-7 tumors 30 minutes after injection. These results suggest that the PEG-peptide-18F-TMR probe displays high selectivity for imaging MMP activity. Conclusions: This strategy successfully images MMP expression in vivo and may be extended to other proteases to predict patient prognosis and to design personalized, protease-based inhibitors and prodrug-targeted therapies.

UR - http://www.scopus.com/inward/record.url?scp=84855423104&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84855423104&partnerID=8YFLogxK

U2 - 10.1158/1078-0432.CCR-11-0608

DO - 10.1158/1078-0432.CCR-11-0608

M3 - Article

VL - 18

SP - 238

EP - 247

JO - Clinical Cancer Research

JF - Clinical Cancer Research

SN - 1078-0432

IS - 1

ER -