Rosas Jonah M, Campanale Joseph P, Harwood Jacob L, Li Lufei, Bae Rachel, Cheng Shujun, Tsou Julia M, Kaiser Kathi M, Engle Dannielle D, Montell Denise J, Pitenis Angela A
Department of Biomolecular Science & Engineering Program, University of California, Santa Barbara, California 93106, United States.
Department of Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, California 93106, United States.
ACS Appl Bio Mater. 2024 Dec 16;7(12):8489-8502. doi: 10.1021/acsabm.4c01301. Epub 2024 Nov 22.
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 two-dimensional (2D) and three-dimensional (3D) environments. However, a 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 morphogenesis and lumen expansion. In confinement, tumor organoids (mT) formed a lumen that expanded rapidly, whereas normal organoids (mN) expanded more slowly. Moreover, a majority of normal organoids in more-confined conditions exhibited an 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 the microgels in the direction of organoid expansion. Normal organoids more commonly exhibit 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 the confinement of healthy cells within a fibrotic tumor niche disrupts tissue organization and function in vivo.
胰腺导管腺癌（PDAC）是一种起源于构成胰腺导管网络上皮细胞的癌症。在疾病进展过程中，PDAC肿瘤会募集促进纤维化的成纤维细胞，增加局部组织硬度，并使上皮细胞承受更大的压缩力。先前的体外研究记录了二维（2D）和三维（3D）环境中压缩应力作用下细胞骨架和细胞核的适应性变化。然而，正常和肿瘤来源的导管上皮细胞对生理相关限制的反应比较仍未得到充分研究，尤其是在3D类器官中。在这里，我们使用由基质胶与低屈服应力颗粒微凝胶混合而成的工程化3D微环境来控制限制条件。将正常和肿瘤来源的小鼠胰腺类器官（正常和肿瘤）在这种复合3D环境或纯基质胶中培养48小时，以研究限制对形态发生和管腔扩张的影响。在限制条件下，肿瘤类器官（mT）形成了一个迅速扩张的管腔，而正常类器官（mN）扩张得更慢。此外，与限制较小条件下的正常类器官相比，大多数处于限制较大条件下的正常类器官表现出倒置的顶-基极性。肿瘤类器官表现出一种集体“脉动”行为，在限制条件下这种行为会增强。这些脉动产生的力足以在类器官扩张方向上局部克服微凝胶的屈服应力。正常类器官更常见地表现出单向旋转。我们的体外微凝胶限制平台揭示了类器官中两种不同的集体力产生模式，这可能有助于阐明肿瘤与微环境之间的相互作用。这些对体外动力学的见解可能会加深我们对纤维化肿瘤微环境中健康细胞的限制如何破坏体内组织结构和功能的理解。
