This project utilizes patient-derived pancreatic ductal adenocarcinoma (PDAC) organoids co-cultured with cancer-associated fibroblasts (CAFs) and immune cells in advanced 3D systems including bioprinted extracellular matrices, organ-on-chip platforms, and decellularized scaffolds to dissect chemotherapy resistance mechanisms driven by dynamic tumor microenvironment (TME) remodeling. Doctoral researchers will establish PDO biobank lines from therapy-naïve and resistant PDAC patients (FOLFIRINOX, nab-paclitaxel/gemcitabine responders/non-responders), perform longitudinal co-culture assays tracking stromal polarization (myCAF/iCAF subtypes), extracellular matrix deposition, and immune cell infiltration under chemotherapy pressure, and apply single-cell RNA sequencing, spatial multiomics, cytokine profiling, and high-throughput drug screening to map TME-mediated resistance networks. Functional validation through CRISPR perturbation of key ligand-receptor axes, secretome analysis, and patient-correlated validation identifies subgroup-specific biomarkers and therapeutic vulnerabilities, enabling development of TME-targeted combination strategies to overcome PDAC chemoresistance.