Patient-Derived Colorectal Cancer Extracellular Matrices Modulate Cancer Cell Stemness Markers

Although it has been shown that the tumor extracellular matrix (ECM) may sustain the cancer stem cell (CSC) niche, its role in the modulation of CSC properties remains poorly characterized. To elucidate this, paired tumor and adjacent normal mucosa, derived from colon cancer patients' surgical resections, were decellularized and recellularized with two distinct colon cancer cells, HT-29 or HCT-15. Methods: The matrix impact on cancer stem cell marker expression was evaluated by flow cytometry and qRT-PCR, while transforming growth factor-β (TGF-β) secretion and matrix metalloprotease (MMP) activity were quantified by ELISA and zymography. Results: In contrast to their paired normal counterparts, the tumor decellularized matrices enhanced HT-29 expression of the pluripotency and stemness genes <i>NANOG</i> (<i>p</i> = 0.0117), <i>SOX2</i> (<i>p</i> = 0.0156), and <i>OCT4</i> (<i>p</i> = 0.0312) and of the epithelial-to-mesenchymal transition (EMT)-associated transcription factor <i>SNAI1</i> (<i>p</i> = 0.0156). Notably, no significant differences were found in the expression of <i>SLUG</i> or <i>TGFB</i> on HT-29 or of the six transcripts on HCT-15 cells. HT-29 mRNA alterations were followed by enhanced expression of the stemness-associated receptors cluster of differentiation 44 (CD44), CD133, and CD166 (<i>p</i> = 0.0078), the secretion of TGF-β (<i>p</i> = 0.0286), and MMP-2 (<i>p</i> = 0.0081) and MMP-9 (<i>p</i> = 0.0402) proteolysis. To infer the clinical relevance of these findings, we assessed cohort databases and evidenced that patients expressing higher levels of the four stemness-associated genes (<i>NANOG</i>/<i>SOX2</i>/<i>OCT4</i>/<i>SNAI1</i>) had worse overall survival. This study demonstrates that normal and tumor matrices harbor different stemness potential and suggest patient-derived decellularized matrices as an excellent three-dimensional (3D) model to unveil stemness signatures, appointing candidates for future therapeutic strategies.