(F-206) Engineering a high throughput multilayer multicellular model of endometrial cancer for drug screening and drug delivery applications

Introduction:: Endometrial cancer is a prevalent gynecological cancer worldwide, with no specific treatments available despite an estimated 382,000 new cases and 90,000 deaths annually. Three-dimensional (3D) in vitro models offer a promising approach for understanding how endometrial cancer metastasizes and screening potential drug candidates. This study aimed to develop, validate, and apply a 3D in vitro model of endometrial cancer that maximized both endometrial cancer invasion and endothelial cell microvessel formation, two key steps in endometrial cancer metastasis. We then used this platform to evaluate the effect of chemotherapy delivery modality on cell survival, cancer invasion, and microvessel formation in 2D and 3D across three different endometrial cancer cell lines.

Materials and Methods:: Multilayer multicellular constructs were fabricated in specialized 96-well plates using human microvascular endothelial cells (hMVECs) and a human endometrial cancer cell line (HEC-1-A) (Fig. 1A, B). Using a Design of Experiments statistical optimization approach, we identified the specific concentrations of natural (collagen type I and IV, fibrinogen, fibronectin, laminin) and synthetic (GELMA, PEGDA) polymers in both hydrogel layers that maximized endometrial cancer invasion and microvessel formation simultaneously. The optimized constructs were then made with two additional endometrial cancer cell lines (Ishikawa and KLE) and exposed to a clinically relevant chemotherapy agent (paclitaxel), either as a free drug or encapsulated in polycaprolactone nanoparticles (Fig. 1C).


Results, Conclusions, and Discussions:: Results. All of the hydrogel components evaluated significantly influenced both cell types in each hydrogel layer (p ≤ 0.05). Moreover, we observed significantly increased endometrial cancer invasion and microvessel formation using our optimized constructs compared to Matrigel, the industry gold standard for 3D cancer models. Using our optimized constructs, we were able to observe the effects of paclitaxel as either a free drug or delivered in a nanoparticle on cancer invasion, microvessel formation, and cell viability which would not be possible with currently available 2D or 3D platforms (Fig. 1D, E).

Conclusion. Our 3D in vitro multilayer multicellular model captured differences among cell lines and captured clinically relevant metastatic behavior in response to different treatment modalities. These platforms can be used to evaluate thousands of compounds simultaneously, empowering large-scale mechanistic studies to investigate the onset and progression of endometrial cancer as well as enable drug discovery and personalized medicine.


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