In Vitro Pancreatic Tumor Microenvironments using DNA-Directed Patterning

As one of the most lethal malignancies worldwide, pancreatic cancer underscores the urgent need for advancements in effective treatment. The pancreatic tumor microenvironment (TME) plays a crucial role in tumor progression, metastasis and immunosuppression, making it an important target for pancreatic cancer research. Among the various techniques applied to explore TME dynamics, DNA-directed patterning stands out as a promising approach through leveraging photolithography-based templating and the unique properties of DNA and Watson-Crick base pairing to engineer intricate cellular arrangements at the microscale. This technique involves two distinct steps: initial patterning of a substrate with short single-stranded oligonucleotides; followed by cell labeling with complementary oligonucleotides and their placement onto the patterned substrate, thus generating precise cellular configurations. These patterns may contain various components of the TME, such as cancer-associated fibroblasts (CAFs), different macrophage phenotypes, and tumor cell nests. Subsequently, by introducing different mixtures of extracellular matrix (ECM) components, researchers can further mimic the native tumor microenvironment with excellent precision. This modulated spatial organization and iterative nature enables investigations into cellular behaviors, including migration, proliferation, differentiation, and responses to therapeutic agents. DNA-directed patterning thus provides a versatile and high-throughput platform to study cellular behaviors in response to varying cues, allowing for a more accurate representation of in vivo conditions.