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Updated: Jun 21, 2025
Quantitative Measurement of Invadopodia-mediated Extracellular Matrix Proteolysis in Single and Multicellular Contexts
Published on: August 27, 2012
Cancer cell extravasation requires iplectin-mediated delivery of MT1-MMP at invadopodia
Olivia R Grafinger1, John J Hayward2, Ying Meng3,4
1Biological Sciences Platform, Sunnybrook Research Institute, Toronto, ON, Canada.
Background:
Invadopodia facilitate cancer cell extravasation, but the molecular mechanism whereby invadopodia-specific proteases such as MT1-MMP are called to invadopodia is unclear.
Methods:
Mass spectrometry and immunoprecipitation were used to identify interactors of MT1-MMP in metastatic breast cancer cells. After identification, siRNA and small molecule inhibitors were used to assess the effect these interactors had on cellular invasiveness. The chicken embryo chorioallantoic membrane (CAM) model was used to assess extravasation and invadopodia formation in vivo.
Results:
In metastatic breast cancer cells, MT1-MMP was found to associate with plectin, a cytolinker and scaffolding protein. Complex formation between plectin and MT1-MMP launches invadopodia formation, a subtype we termed iplectin (i = invadopodial). iPlectin delivers MT1-MMP to invadopodia and is indispensable for regulating cell surface levels of the enzyme. Genetic depletion of plectin with siRNA reduced invadopodia formation and cell invasion in vitro. In vivo extravasation efficiency assays and intravital imaging revealed iplectin to be a key contributor to invadopodia ultrastructure and essential for extravasation. Pharmacologic inhibition of plectin using the small molecule Plecstatin-1 (PST-1) abrogated MT1-MMP delivery to invadopodia and extravasation efficiency.
Conclusions:
Anti-metastasis therapeutic approaches that target invadopodia are possible by disrupting interactions between MT1-MMP and iplectin.
Clinical Trial Registration Number:
NCT04608357.
Insights
Plectin delivers MT1-MMP to invadopodia, facilitating cancer cell invasion and extravasation. Targeting the MT1-MMP and plectin interaction may offer new anti-metastasis therapies.
Area of Science:
- Oncology
- Cell Biology
- Biochemistry
Background:
- Invadopodia are crucial for cancer cell extravasation, but the recruitment mechanism of proteases like MT1-MMP to invadopodia remains unclear.
- Understanding these mechanisms is vital for developing anti-metastasis strategies.
Purpose of the Study:
- To elucidate the molecular mechanism of MT1-MMP recruitment to invadopodia.
- To identify key proteins interacting with MT1-MMP in metastatic breast cancer cells.
Main Methods:
- Mass spectrometry and immunoprecipitation identified MT1-MMP interactors.
- siRNA and small molecule inhibitors assessed effects on invasiveness.
- The chicken embryo chorioallantoic membrane (CAM) model evaluated in vivo extravasation and invadopodia formation.
Main Results:
- Plectin was identified as an MT1-MMP interactor, forming a complex termed 'iPlectin' that drives invadopodia formation.
- iPlectin is essential for MT1-MMP delivery to invadopodia and regulates its cell surface levels.
- Plectin depletion or inhibition (Plecstatin-1) reduced invadopodia formation, invasion, and in vivo extravasation.
Conclusions:
- Disrupting the MT1-MMP and iPlectin interaction presents a potential therapeutic strategy against cancer metastasis.
- Targeting invadopodia-mediated processes offers a novel approach for anti-metastasis treatments.
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