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Noninvasive Bioluminescence Imaging of Matrix Metalloproteinase-14 Activity in Lung Cancer Using a Membrane-Bound
Feng Tian1, Yan Chen2, Wuping Wang1
1Department of Thoracic Surgery, Tangdu Hospital, Air Force Medical University, No. 569 Xinsi Road, Xi'an 710038, China.
Abstract:
Matrix metalloproteinase-14 (MMP-14) plays a crucial role in the cancer migration and metastasis by guiding the extracellular matrix remodeling and cell motility. Despite increasing efforts have been taken to develop methodology for measuring MMP-14 expression, there is a lack of tools capable of monitoring the MMP-14 dynamic activity with high temporal and spatial resolution in living cells and animals. Here, we describe the design of Gaussia luciferase (Gluc)-based membrane-bound biosensor for efficient visualization of MMP-14 activity. The epidermal growth factor (EGF) induced significant luciferase changes in the biosensor-transfected lung cancer cells. Deletion of the transmembrane domain in the mutant biosensor or treatment with an MMP-14 inhibitor, tissue inhibitor of metalloproteinase-2 (TIMP-2), relieved the EGF-induced luciferase activation, suggesting that MMP-14 functions at the cell surface to result in luciferase changes. Moreover, utilizing this biosensor, the bioluminescence signals activated by MMP-14 enabled clear visualization of MMP-14-positive lung tumors in animal models. Our results indicated this biosensor is an effective probe for quantitatively monitoring proteolytic activities in live cells and mouse models. These findings offer the general design of biosensors as an adaptable tool for studying various membrane-anchored proteases in biological models.
Insights
We developed a novel Gaussia luciferase-based biosensor to visualize matrix metalloproteinase-14 (MMP-14) activity in real-time. This tool enables effective monitoring of MMP-14 in cancer cells and animal models, aiding metastasis research.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- Matrix metalloproteinase-14 (MMP-14) is vital for cancer cell migration and metastasis through extracellular matrix remodeling.
- Existing methods lack the temporal and spatial resolution to monitor MMP-14 activity in living systems.
Purpose of the Study:
- To design and validate a Gaussia luciferase (Gluc)-based membrane-bound biosensor for visualizing MMP-14 activity.
- To assess the biosensor's efficacy in monitoring MMP-14 in cancer cells and animal models.
Main Methods:
- Constructed a Gluc-based membrane-bound biosensor for MMP-14.
- Utilized lung cancer cells transfected with the biosensor and stimulated with epidermal growth factor (EGF).
- Tested biosensor response with a mutated biosensor lacking a transmembrane domain and with an MMP-14 inhibitor (TIMP-2).
- Visualized MMP-14-positive lung tumors in animal models using bioluminescence.
Main Results:
- EGF stimulation induced significant luciferase changes in biosensor-transfected cells.
- MMP-14 activity at the cell surface was confirmed to modulate luciferase levels.
- The biosensor successfully visualized MMP-14-positive lung tumors in vivo.
- The biosensor provides quantitative monitoring of proteolytic activity in live cells and animal models.
Conclusions:
- The developed Gluc-based biosensor effectively visualizes MMP-14 activity with high spatiotemporal resolution.
- This biosensor is a valuable tool for studying cancer metastasis and MMP-14 function.
- The biosensor design is adaptable for studying other membrane-anchored proteases.
