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Dryad

Differential Effects of Confinement on the Dynamics of Normal and Tumor-Derived Pancreatic Ductal Organoids

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Dec 04, 2024 version files 474.64 KB

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is a cancer of the epithelia comprising the ductal network of the pancreas. During disease progression, PDAC tumors recruit fibroblasts that promote fibrosis, increasing local tissue stiffness and subjecting epithelial cells to increased compressive forces. Previous in vitro studies have documented cytoskeletal and nuclear adaptation following compressive stresses in 2D and 3D. However, comparison of the responses of normal and tumor-derived ductal epithelia to physiologically relevant confinement remains underexplored, especially in 3D organoids. Here we control confinement with an engineered 3D microenvironment composed of Matrigel® mixed with a low yield stress granular microgel. Normal and tumor-derived murine pancreas organoids (normal and tumor) were cultured for 48 h within this composite 3D environment or in pure Matrigel® to investigate the effects of confinement on lumen morphogenesis. In confinement, tumor organoids (mT) formed lumen that expanded rapidly, whereas normal organoids (mN) expanded more slowly. Moreover, normal organoids in more confined conditions exhibited inverted apicobasal polarity compared to those in less confined conditions. Tumor organoids exhibited a collective “pulsing” behavior that increased in confinement. These pulses generated forces sufficient to locally overcome the yield stress of microgels in the direction of organoid expansion. Normal organoids more commonly exhibited unidirectional rotation. Our in vitro microgel confinement platform enabled the discovery of two distinct modes of collective force generation in organoids that may shed light on the mutual interactions between tumors and the microenvironment. These insights into in vitro dynamics may deepen our understanding of how confinement of healthy cells within a fibrotic tumor niche disrupts tissue organization and function in vivo.