Engineering a three-dimensional multilayer multicellular model of endometrial cancer for high throughput drug screening and novel treatment methods

Abstract

ABSTRACTEndometrial cancer is one of the most common gynecological cancers in the world, with an estimated 382,000 new cases and 90,000 deaths each year. There is no specific treatment, as the underlying causes of endometrial cancer neoplasia are poorly understood. This study focuses on the development and validation of a three-dimensional (3D)in vitromultilayer, multicellularhydrogel that facilitates drug screening analysis. We hypothesized that a specific combination of natural (collagen type I and IV, fibrinogen, fibronectin, Laminin) and synthetic (GELMA, PEGDA) polymers would maximize microvessel formation and cell invasion. The 3D model incorporates human microvascular endothelial cells (hMVEC) and endometrial cancer cells (HEC-1A) atop hydrogel formulations mimicking cell-specific extracellular matrix components. Using a D-optimal experimental design, 45 hydrogel combinations were generated. The predicted hydrogel formulation to maximize all cell responses enhanced higher microvessel formation and cancer invasion compared to the gold standard Matrigel. Subsequent validation emphasizes the importance of a disease-specific model and cell crosstalk in maximizing microvessel formation and cancer invasion. The optimized 3D model adeptly captures variances in cell responses among endometrial cancer cell lines from distinct stages. Finally, the platform is employed to compare cell viability, microvessel formation, and cancer invasion across Ishikawa, KLE, and HEC-1A cells after Paclitaxel exposure, delivered both as a free drug and loaded in poly(caprolactone) (PCL) nanoparticles. Overall, this study provides a valuable tool for exploring intricate interactions within the tumor microenvironment, offering a holistic understanding of cell responses and fostering the development of targeted therapeutic strategies for endometrial cancer.