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Nanosensors to Detect Protease Activity In Vivo for Noninvasive Diagnostics
Published on: July 16, 2018
MMP-9 activity in urine from patients with various tumors, as measured by a novel MMP activity assay using modified
R Hanemaaijer1, C F Sier, H Visser
1Gaubius Laboratory, TNO-PG, Leiden, The Netherlands. R.Hanemaaijer@PG.TNO.NL
Abstract:
Matrix metalloproteinases (MMPs) play an important role in many pathologic processes, but their activities are difficult to determine since no simple specific and/or chromogenic substrate exists. We have developed a novel MMP activity assay using a modified urokinase as a substrate. Protein engineering enabled the plasmin activation site in this urokinase to be substituted by a specific activation site recognized by MMPs. In this way the MMP activity can be monitored via urokinase activity as measured by a simple chromogenic assay. The assay was made specific for MMP-9 by a capture step using MMP-9-specific antibodies that do not interfere with MMP-activity. This assay monitors the amount of active enzyme as well as the latent, but potentially active proform. Using this assay the levels of MMP-9 were investigated in urine from patients with various kinds of carcinoma. High levels of both active and latent MMP-9 were detected in urine from patients with carcinoma of the bladder, whereas hardly any activity was observed in urine from healthy controls. MMP-9 in urine was present in its intact form. Surprisingly, MMP-9 was also increased in the urine of patients with nonurogenital carcinoma. Therefore, measurement of urinary MMP-9 activity levels may be a convenient diagnostic tool for various types of carcinoma.
Insights
Researchers developed a novel assay to measure matrix metalloproteinase-9 (MMP-9) activity in urine. Elevated MMP-9 levels were detected in patients with various carcinomas, suggesting its potential as a diagnostic tool.
Area of Science:
- Biochemistry
- Enzymology
- Oncology
Background:
- Matrix metalloproteinases (MMPs) are crucial in pathological processes.
- Assessing MMP activity is challenging due to the lack of specific substrates.
- Matrix metalloproteinase-9 (MMP-9) is implicated in various diseases.
Purpose of the Study:
- To develop a novel, specific assay for measuring MMP activity.
- To specifically detect and quantify matrix metalloproteinase-9 (MMP-9) activity in biological samples.
- To investigate the potential of urinary MMP-9 as a biomarker for carcinoma.
Main Methods:
- Engineered urokinase as a substrate with an MMP-specific activation site.
- Developed a chromogenic assay to monitor urokinase activity, reflecting MMP activity.
- Incorporated MMP-9-specific antibodies for targeted detection and quantification of MMP-9.
- Analyzed MMP-9 levels in urine samples from carcinoma patients and healthy controls.
Main Results:
- The novel assay successfully detected both active and latent forms of MMP-9.
- Significantly elevated levels of active and latent MMP-9 were found in the urine of bladder carcinoma patients compared to healthy individuals.
- Increased MMP-9 levels were also observed in patients with nonurogenital carcinomas.
- Urinary MMP-9 was predominantly detected in its intact form.
Conclusions:
- The developed assay is a sensitive tool for measuring MMP-9 activity.
- Urinary MMP-9 levels are significantly elevated in patients with various carcinomas.
- Measurement of urinary MMP-9 activity shows promise as a convenient diagnostic biomarker for carcinoma detection.