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Translational Therapeutics

Cancer cell extravasation requires iplectin-mediated delivery of MT1-MMP at invadopodia

British Journal of Cancer volume 131, pages 931–943 (2024)Cite this article

Subjects

Abstract

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

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Fig. 1: Plectin is highly expressed in human breast cancer and associates with MT1-MMP.
Fig. 2: Plectin is required for invadopodia formation and function through the recruitment of MT1-MMP.
Fig. 3: Plectin knockdown abrogates extravasation potential formation in vivo.
Fig. 4: Plecstatin-1 alters cytoskeletal networks and consequently disrupts the delivery of MT1-MMP to F-actin puncta.
Fig. 5: The plectin antagonist, plecstatin-1, is a potent inhibitor of invadopodia formation and cell invasion in multiple cell types.

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Data availability

The .RAW and affiliated files were deposited into the publicly available PRIDE partner database for the ProteomeXchange consortium with the data set identifier: PXD039276.

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Acknowledgements

We thank Ms. Lilianne Gee for assisting with sample preparation for mass spectrometry analysis and Dr. Jonathan Kriger of Bioinformatics Solutions Inc. for operation of the mass spectrometer.

Funding

HSL and KJJ are supported by a Kavelman-Fonn Foundation Operating Grant. HSL received support from CIHR (Project Grant 480369) and a Movember/PCC grant (PC-RS16). MGC received support from NSERC (Discovery Grant 05199). JFT received support from NSERC (Discovery Grant 06338).

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Authors and Affiliations

  1. Biological Sciences Platform, Sunnybrook Research Institute, Toronto, ON, Canada

    Olivia R. Grafinger, Yan Li, Sara Mar, Minzhi Sheng, Boyang Su, Gobi Thillainadesan & Hon S. Leong

  2. Department of Chemistry, University of Windsor, Windsor, ON, Canada

    John J. Hayward & John F. Trant

  3. Harquail Centre for Neuromodulation, Sunnybrook Health Sciences Centre, Toronto, ON, Canada

    Ying Meng & Nir Lipsman

  4. Division of Neurosurgery, Sunnybrook Health Sciences Centre, University of Toronto, Toronto, ON, Canada

    Ying Meng & Nir Lipsman

  5. Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada

    Jennifer Geddes-McAlister & Marc G. Coppolino

  6. Department of Medical Biophysics, Temerty Faculty of Medicine, University of Toronto, Toronto, ON, Canada

    Sara Mar, Minzhi Sheng, Boyang Su, Katarzyna J. Jerzak & Hon S. Leong

  7. Division of Medical Oncology, Sunnybrook Odette Cancer Centre, University of Toronto, Toronto, ON, Canada

    Katarzyna J. Jerzak

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  1. Olivia R. Grafinger
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Contributions

ORG performed experiments, collected and analyzed data, prepared figures, and drafted the manuscript. JJH performed experiments, collected and analyzed data, prepared figures, and edited the manuscript. YM assisted in the establishment of the Primary-BM cell line. JG-M planned experiments and performed data analysis for mass spectrometry profiling. SM prepared and maintained avian embryos. YL performed experiments. MS performed experiments. BS assisted in cell culture and establishment of the zsGreen-MDA-MB-231 and zsGreen-Primary-BM cell lines. GT established the Primary-BM cell line. NL assisted in the establishment of the Primary-BM cell line. JFT planned experiments, drafted, and edited the manuscript. MGC planned experiments. KJJ assisted in the establishment of the Primary-BM cell line. HSL planned experiments, drafted, and edited the manuscript.

Corresponding author

Correspondence to Hon S. Leong.

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Patients were consented for use of their tumor tissue with Sunnybrook REB approval (SUN-2047) for NCT04608357. The study was performed in accordance with the Declaration of Helsinki.

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Grafinger, O.R., Hayward, J.J., Meng, Y. et al. Cancer cell extravasation requires iplectin-mediated delivery of MT1-MMP at invadopodia. Br J Cancer 131, 931–943 (2024). https://doi.org/10.1038/s41416-024-02782-9

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