Novel 3D µtissues Mimicking the Fibrotic Stroma in Pancreatic Cancer to Study Cellular Interactions and Stroma-Modulating Therapeutics

SIMPLE SUMMARY: Pancreatic ductal adenocarcinoma (PDAC) is the most prevalent and aggressive type of pancreatic cancer with a low 5-year survival rate of only 8%. The cellular arrangement plays a crucial role in PDAC, which is characterized by a highly fibrotic environment around the tumor cells, preventing treatments from reaching their target. For the development of novel drug candidates, it is crucial to mimic this cellular arrangement in a laboratory environment. We successfully developed a reproducible three-dimensional cell culture model that demonstrates the PDAC characteristic arrangement and showed a PDAC relevant gene profile when comparing with the genetic profile of PDAC patients. We finally demonstrated the use of the model for the evaluation of novel anti-fibrotic therapy against PDAC by studying drug-induced reduction of fibrosis in PDAC enabling nanoparticles to penetrate and reach the tumor cells. This model is useful for the evaluation of novel treatments against PDAC in a biologically relevant manner. ABSTRACT: Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive tumor type with low patient survival due to the low efficacy of current treatment options. Cancer-associated fibroblasts (CAFs) in the tumor microenvironment (TME) create a dense fibrotic environment around the tumor cells, preventing therapies from reaching their target. Novel 3D in vitro models are needed that mimic this fibrotic barrier for the development of therapies in a biologically relevant environment. Here, novel PDAC microtissues (µtissues) consisting of pancreatic cancer cell core surrounded by a CAF-laden collagen gel are presented, that is based on the cells own contractility to form a hard-to-penetrate barrier. The contraction of CAFs is demonstrated facilitating the embedding of tumor cells in the center of the µtissue as observed in patients. The µtissues displayed a PDAC-relevant gene expression by comparing their gene profile with transcriptomic patient data. Furthermore, the CAF-dependent proliferation of cancer cells is presented, as well as the suitability of the µtissues to serve as a platform for the screening of CAF-modulating therapies in combination with other (nano)therapies. It is envisioned that these PDAC µtissues can serve as a high-throughput platform for studying cellular interactions in PDAC and for evaluating different treatment strategies in the future.