Abstract：
The tumor microenvironment (TME) orchestrates cellular and extracellular matrix (ECM) interactions, playing a key role in tumorigenesis, tumor growth, and metastization. Investigating the interplay between stromal-epithelial cells within the TME is paramount for understanding cancer mechanisms but demands reliable biological models. 3D-models have emerged as powerful in vitro tools, but many fall short in replicating cell-cell/cell-matrix interactions. This study introduces a novel hybrid 3D-model of the breast TME, combining epithelial cells, cancer-associated fibroblasts (CAFs), and their ECM. To build the stromal compartment, porous 3D-printed alginate scaffolds were seeded with CAFs, which proliferated and produced ECM. The pores were infused with oxidized peptide-modified alginate hydrogel laden with MCF10A cells, forming the parenchymal compartment. The hybrid system supported epithelial morphogenesis into acini surrounded by fibroblasts and ECM, and could be readily solubilized to recover cells, their matrix, and sequestered soluble factors. Proteome profiling of the retrieved ECM showed upregulation of proteins associated with matrix assembly/remodeling, epithelial-to-mesenchymal transition (EMT), and cancer. The TME-like microenvironment induced a partial EMT in MCF10A cells, generating a hybrid population with epithelial and mesenchymal features, characteristic of aggressive phenotypes. Our model provided new insights into epithelial-stromal interactions within the TME, offering a valuable tool for cancer research in a physiologically-relevant 3D setting.
摘要：
肿瘤微环境（TME）协调细胞和细胞外基质（ECM）相互作用，在肿瘤发生中起关键作用，肿瘤生长，和转化。研究TME内基质上皮细胞之间的相互作用对于理解癌症机制至关重要，但需要可靠的生物学模型。3D模型已经成为强大的体外工具，但是许多在复制细胞-细胞/细胞-基质相互作用方面不足。这项研究介绍了一种新颖的乳房TME的混合3D模型，结合上皮细胞，癌症相关成纤维细胞（CAFs），和他们的ECM。为了建造基质隔间，多孔3D打印的藻酸盐支架接种CAF，增殖并产生ECM。用载有MCF10A细胞的氧化肽修饰的藻酸盐水凝胶注入孔，形成实质室。杂合系统支持上皮形态发生，进入由成纤维细胞和ECM包围的腺泡，并且可以很容易地溶解以回收细胞，他们的矩阵，和螯合的可溶性因子。回收的ECM的蛋白质组分析显示与基质组装/重塑相关的蛋白质上调，上皮-间质转化（EMT），和癌症。TME样微环境在MCF10A细胞中诱导部分EMT，产生具有上皮和间质特征的杂种群体，侵袭性表型的特征。我们的模型为TME内的上皮-基质相互作用提供了新的见解，在生理相关的3D环境中为癌症研究提供了有价值的工具。
