An Engineered Paper-Based 3D Co-Culture Model of Pancreatic Cancer as a Platform for Systems Tissue Engineering

Pancreatic ductal adenocarcinoma (PDAC) is a deadly form of pancreatic cancer and is projected to be the third leading cause of cancer-related deaths in Canada this year. With a 5-year survival rate that has barely improved over the last decade, new therapies are sorely needed to treat this disease. PDAC tumours are characterized by a stroma-rich and hypoxic tumour microenvironment (TME) that drives disease progression and results in a highly aggressive disease. Furthermore, the spatial configuration of cells in the TME affects cell phenotypes, which contributes to the clinical challenge of tumour heterogeneity and therapeutic resistance. Yet, pre-clinical studies typically rely on growing cancer cells deprived of their native complex and heterogeneous TME. Patient-derived organoids (PDOs) are emerging as powerful tools to model cancer, but their self-assembling nature and lack of stromal cells results in microenvironments that are not controllable, nor physiological. The principles of tissue engineering provide a powerful toolbox to elevate PDO cultures to better mimic the complex TME of PDAC. In this work, I present a versatile engineered 3D in vitro stroma-cancer co-culture model of PDAC based on the Tissue Roll for Analysis of Cellular Environment and Response (TRACER). By leveraging patterned polymer infiltration to reduce the number of cells required for TRACER, we incorporate human PDOs into our system. PDOs establish oxygen gradients across TRACER and in response exhibit graded cell viability, proliferation, hypoxia-response gene expression, and response to chemotherapy. We then incorporate primary pancreatic stellate cells (PSCs) to produce co-culture tissues with user-defined architectures that were inferred from clinical PDAC samples. By performing high dimensional characterization using CyTOF, we demonstrate that tissue architecture leads to distinct hypoxia and proliferation gradients. Furthermore, phenotypic markers for both cell types are also graded in ways that cannot be explained by either hypoxia or co-culture alone. I anticipate that our engineered tissue model will enable systems biology studies not possible with classic in vitro models. Specifically, our model will help decipher how tissue architecture and cell interactions regulate cell phenotype and, in turn, tissue-scale phenotypes that can be targeted for therapeutic purposes.

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