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Multicompartmentalized microvascularized tumor-on-a-chip to study tumor-stroma interactions and drug resistance in ovarian cancer

Authors: Simona Plesselova, Kristin Calar, Hailey Axemaker, Emma Sahly, and Pilar de la Puente

Abstract

The majority of ovarian cancer (OC) patients receiving standard of care chemotherapy develop chemoresistance within 5 years. The tumor microenvironment (TME) is a dynamic and influential player in disease progression and therapeutic response. However, there is a lack of models that allow us to elucidate the compartmentalized nature of TME in a controllable, yet physiologically relevant manner and its critical role in modulating drug resistance. We developed a 3D microvascularized multiniche tumor-on-a-chip formed by five chambers (central cancer chamber, flanked by two lateral stromal chambers and two external circulation chambers) to recapitulate OC-TME compartmentalization and study its influence on drug resistance. Stromal chambers included endothelial cells alone or cocultured with normal fibroblasts or cancer-associated fibroblasts (CAF). The tumor-on-a-chip recapitulated spatial TME compartmentalization including vessel-like structure, stromal-mediated extracellular matrix (ECM) remodeling, generation of oxygen gradients, and delayed drug diffusion/penetration from the circulation chamber towards the cancer chamber. The cancer chamber mimicked metastasis-like migration and increased drug resistance to carboplatin/paclitaxel treatment in the presence of CAF when compared to normal fibroblasts. CAF-mediated drug resistance was rescued by ECM targeted therapy. Critically, these results demonstrate that cellular crosstalk recreation and spatial organization through compartmentalization are essential to determining the effect of the compartmentalized OC-TME on drug resistance. Our results present a functionally characterized microvascularized multiniche tumor-on-a-chip able to recapitulate TME compartmentalization influencing drug resistance. This technology holds the potential to guide the design of more effective and targeted therapeutic strategies to overcome chemoresistance in OC.


Fig. Multiniche microvascularized tumor-on-a-chip recapitulates the compartmentalized OC-TME. (A) Schematic representation of tumor-on-a-chip device and close-up view of the different chambers and cell types seeded inside (left) and real image of microfluidic device filled with dyes next to a nickel representing the size of the platform (right). (B) Representative images of the stromal chamber with vessel-like structures formation of endothelial cells (EC) stained with BV605-anti-CD31 (red) in monoculture or coculture with normal fibroblasts (NF) or cancerassociated fibroblasts (CAF) stained with FITC-anti-CD90 (green) and the quantification of the circularity, area, inner diameter, and elongation of the vessels. Median value is marked in red. Scale Bar = 100μm. *p<0.05, One-Way ANOVA compared to EC.

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Keywords: tumor-on-a-chip; compartmentalization; ovarian cancer; drug resistance; tumor microenvironment; microfluidics; reservoir connector

bioRxiv 2024

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