This project analyzes acinar-to-ductal metaplasia (ADM) progression and real-time transcription factor kinetics in fresh pancreatic explants and patient-derived organoids using single-cell multiomics and advanced live-cell imaging. Doctoral researchers will establish organoid cultures from normal pancreas, ADM lesions, and early PDAC precursors, engineer reporter lines for key ADM drivers (e.g., SOX9, TP63, KLF5, HNF1B), and perform longitudinal single-cell RNA-seq, ATAC-seq, and single-molecule tracking (e.g., RBPJ-DNA binding kinetics via HaloTag/dSTORM) to map dynamic transcriptional networks during ADM onset, progression to PanIN, and early invasion. Integration with spatial transcriptomics on pancreatic slices and core facility proteomics will reveal epigenetic remodeling, lineage plasticity states, and cell-of-origin effects distinguishing ADM from malignant transformation. Functional CRISPR screens and perturbation experiments validate critical regulatory hubs, identifying biomarkers for early PDAC interception and therapeutic windows to block ADM-driven oncogenesis before invasive disease establishment.