Speaker
Description
Introduction
Pancreatic Ductal Adenocarcinoma (PDAC) is hallmarked by a dense, collagen-rich stroma, driven by the activation of pancreatic stellate cells (PSCs), which remodel the extracellular matrix (ECM) into a mechanically stiff microenvironment that promotes tumor progression and therapeutic resistance.1,2 To accurately mimic this fibrotic transformation in vitro, we engineered 3D scaffolds using type I collagen — the predominant ECM protein in PDAC — as a tunable biomaterial. Collagen's native bioactivity, cell-binding motifs, and mechanical adaptability make it a powerful tool for biofabricating physiologically relevant tumor microenvironments.3,4
Methods
Immortalized human PSCs behavior was investigated in co-culture with two different immortalized human pancreatic cancer cell lines (AsPC-1 or PANC-1) and compared to the stromal monoculture. Three-dimensional constructs consisting of cells embedded within collagen matrices were prepared varying the collagen concentration from 0.5 to 2.5 mg/mL.
After 48h of culture, activation of the stromal component was studied through the expression of alpha-smooth muscle actin (α-SMA), while N-cadherin and E-cadherin were used as cancer epithelial-to-mesenchymal transition (EMT) markers. Such protein expressions were investigated by immunofluorescence and quantitatively by Western Blot. Moreover, samples stained with DAPI and phalloidin were analyzed to measure variations in cell nuclei morphology and arrangement of the cytoskeleton, respectively, comparing co-cultures of stromal and cancer cells with the single populations.
Results
Hydrogels spanning physiological to pathological stiffnesses induced marked differences in nuclear and cytoskeletal architecture, particularly in stromal cells. PSCs showed stiffness-dependent activation, with α-SMA upregulation in both mono- and co-cultures, more pronounced in AsPC-1 co-cultures. PANC-1 cells acquired EMT markers in stiffer scaffolds, while AsPC-1 displayed inverse N-cadherin trends.
Discussion
This study demonstrates how collagen stiffness modulates stromal and cancer cell behavior in 3D, recapitulating key aspects of PDAC desmoplasia. The observed crosstalk and mechanical feedback highlight the value of engineered ECM scaffolds as predictive platforms for tumor-stroma interactions. Our biofabricated model offers a tunable, physiologically relevant framework to dissect mechano-biological cues in cancer progression.
References
1 MacCurtain BM, et.al., J Clin Med. 2021; 10.
2 Osuna de la Peña D, et al., Nat Commun. 2021; 12.
3 Antoine EE, et al., PLoS One. 2015; 10.
4 Szot CS, et al., Biomaterials. 2011; 32.
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