Poster abstracts

Poster number 41 submitted by Ariana Shannon

Elucidating Protein-based Crosstalk between Epithelial and Fibroblast Cells in a Colon Cancer Cell Model with Mass Spectrometry-based Proteomics

Ariana E. Shannon (Ohio State Biochemistry Program and Ohio State University), Amanda B. Hummon (Chemistry and Biochemistry and Ohio State University)

Abstract:
Once activated by tumor cells, cancer-associated fibroblasts (CAFs) modulate the tumor microenvironment through the release of protein growth factors. We propose using 3-dimensional tumor models cocultured with epithelial cells and activated fibroblasts to tease out the proteomic crosstalk using improved mass spectrometry methods, specifically data-independent acquisition (DIA). When applied to cell culture-based tumor modelling in vitro, recent improvements in DIA and data analysis pipelines may aid the field in moving closer to personalized-precision medicine.

We will test three conditions, with an n=4; cocultures of HCT 116 carcinoma cell line with untreated CCD-18Co fibroblasts, TGF-β1-activated fibroblasts, or tumor cell conditioned-medium treated-fibroblasts. Controls of HCT 116, activated and non-activated fibroblasts will be grown in monoculture.

We anticipate conditioned medium will activate fibroblasts to CAF. Previously, we detected TGF-β1 receptor peptides within monocultured spheroids only. In coculture, significantly downregulated (p>0.05) proteins that participate in or interact with the TGF-β1 signaling were detected using gas phase fractionation (GPF) DIA. Additionally, proteins involved in metabolic changes and the epithelial-to-mesenchymal transition were upregulated, which is consistent with published literature. We intend to use proteomics to understand similarly activated fibroblasts in 2D prior to translating the model into a 3D-construct.

Keywords: coculture, cancer, proteomics