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    Biomaterials

    A Proteomics Based Approach for Material Selection in a Model of the Pre-Metastatic Niche

    (C-115) A Proteomics Based Approach for Material Selection in a Model of the Pre-Metastatic Niche

    Thursday, October 12, 2023
    9:30 AM – 10:30 AM PDT
    Location: Exhibit Hall - Row C - Poster # 115
    Introduction: Breast cancer is the most common cancer and second leading cause of cancer death in females. While the prognosis is good for localized cancers, metastatic breast cancer remains to be a challenge with a five-year survival rate of 29%. This poor prognosis is in part due to a lack of understanding of the pre-metastatic niche and a lack of in vitro models to study its dynamics. Previously, our lab has developed a model of the pre-metastatic niche consisting of a synthetic SU-8 scaffold coated with fibronectin (FN), an extracellular matrix protein which drives ECM remodelling in cancer. However, in developing this model we observed that varying degrees of coating were achieved when different brands of commercially available FN were used, thereby potentially altering the efficacy of the model.

    Materials and Methods: Here, we compared five commercially available human-derived FN products commonly used as substrates for cell culture. FN fibril formation and coating efficiency on three-dimensional tissue scaffolds were evaluated using confocal microscopy. Proteomic analysis was then performed on each product to determine overall protein composition and peptide expression levels for the FN1 gene.

    Results, Conclusions, and Discussions: When analyzing coating efficiency, only two brands of FN produced fibrils on the SU-8 scaffolds, Corning and Sigma. These two brands also had the highest purities and lowest numbers of protein coisolates as determined by LC-MS/MS. Examining peptide expression levels for the FN1 gene demonstrated that domains associated with fibrollogenesis were significantly upregulated in the Sigma and Corning formulations, providing a biological indicator as to why these products were more likely to form fibrils on the synthetic scaffolds. These results illustrate how material selection is crucial for the construction of in vitro models. Future work will include a proteomic comparison of FN secreted by breast cancer cell lines to the commercial formulations outlined here to further determine which product is most representative of the pre-metastatic niche.

    Acknowledgements (Optional): The authors would like to thank METAvivor and the Purdue Proteomics Facility for their support.

    References (Optional):