High metastatic potential and early systemic dissemination are hallmarks of pancreatic adenocarcinoma (PDAC). Despite extensive knowledge of the genetics of PDAC, the molecular mechanisms underlying PDAC’s propensity to invade and metastasise are not well understood. Within this project, we will exploit a unique collection of primary human 3D organoids derived from patients undergoing resection for PDAC at our institution. These organoid cultures preserve genotype and phenotype of individual PDACs and therefore uniquely reflect the genetic and biological heterogeneity of this disease. We will analyse individual tumor’s migratory, invasive and metastatic capacity ex vivo and integrate functional data with comprehensive molecular analyses and clinical data. We aim to establish novel biomarkers of PDAC metastatic capacity that could help stratify patients according to individual tumor biology. We also aim to validate candidate drivers and modulators of PDAC dissemination through mechanistic ex vivo studies in 3D organoids. In recent in silico work, we established a robust classifier that distinguished PDAC from non-tumorous tissues and additionally showed both a diagnostic and also prognostic impact. In this context, we observed a set of genes upregulated in PDAC that have been shown to play a role in cell motility, invasion and metastasis, amongst them Trefoil Factor 1 (TFF1) and G Protein coupled receptor class C group 5 member A (GPRC5A). We will target candidate genes in patient-derived organoids and established cell lines employing RNAi-based downregulation, lentiviral overexpression and CRISP/CAS-mediated genome engineering. Findings will be correlated with clinical data from our patient cohort and validated with published patient cohorts.