Abstract
The extracellular matrix (ECM) is a molecular scaffold mainly comprising fibrous proteins, glycoproteins, proteoglycans, and polysaccharides. Aside from acting as a structural support, the ECM’s composition dictates cell-matrix interactions at the biochemical and biophysical level. In the context of cancer, the ECM is a critical component of the tumor microenvironment (TME) and dysregulation of its deposition and remodelling has been shown to promote tumor onset, progression, and metastasis. Here, we describe a robust protocol for the isolation and subsequent proteomic analysis of the ECM of murine mammary glands, for downstream assays studying the role of the ECM in breast cancer. The protocol yields sufficient protein amounts to enable not only the global quantitation of protein expression but furthermore the enrichment and quantitative analysis of post-translationally modified peptides to study aberrant signalling.
Key words
- Cancer
- Tumor microenvironment (TME)
- Extracellular matrix
- Proteomics
- Signalling
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Acknowledgments
SEJP would like to acknowledge fellowships from Fonds de recherche du Québec (Santé) and the Epstein Fellowship in Women’s Health (Faculty of Medicine, McGill University). SVDR would like to thank the Canadian Institutes of Health Research (CIHR) (grants PJT-162260 and PJT-156269 to SVDR) and the Canadian Cancer Society (grant #707140 to SVDR) for financial support. CHB would like to thank Genome Canada for financial support through the Genomics Technology Platform (GTP: 264PRO), and the Segal McGill Chair in Molecular Oncology at McGill University (Montreal, Quebec, Canada). CHB would also like to thank the Warren Y. Soper Charitable Trust and the Alvin Segal Family Foundation for supporting the Jewish General Hospital (Montreal, Quebec, Canada). RPZ would like to thank the University of Manitoba and Shared Health Manitoba for financial support. The authors are also grateful for financial support from the Terry Fox Research Institute.
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Preston, S.E.J., Richard, V.R., del Rincón, S.V., Borchers, C.H., Zahedi, R.P. (2023). Proteomic Assessment of the Murine Mammary Gland Extracellular Matrix. In: Ursini-Siegel, J. (eds) The Tumor Microenvironment. Methods in Molecular Biology, vol 2614. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2914-7_16
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DOI: https://doi.org/10.1007/978-1-0716-2914-7_16
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2913-0
Online ISBN: 978-1-0716-2914-7
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