Mapping cellular heterogeneity and dynamic interactions in pancreatic cancer

Pancreatic ductal adenocarcinoma (PDAC) is highly aggressive with a high mortality rate. Intra-tumoral heterogeneity (ITH) increases the severity of PDAC and makes treatment difficult. Insights are provided on ITH to understand the diversity of microenvironment (ME) components, biomarkers, different subsets of tumor-associated cells, and immune cells, as well as metabolic reprogramming, autophagy, and apoptosis in PDAC. Single-cell RNA sequencing (scRNA-seq) is a sensitive technique that provides spatially resolved transcriptomic profiling. In this review, we discussed the sample preparation, library preparation, data analysis, and challenges associated with the technology. We have outlined a stepwise process workflow that utilizes computational approaches based on experimental requirements, supported by relevant examples and discussion. We reviewed various studies where scRNA-seq has helped identify dynamic cell subset transformations during tumorigenesis, modulate ME, epithelial-mesenchymal transition, and cancer stem cells enrichment, and identify novel signaling molecules, prognostic gene markers, and therapeutic vulnerabilities for PDAC. Identification of biomarkers such as Matrix metalloproteinase 1 (MMP1) and the S100A2⁺ tumor subset, characterization of the basal-like malignant subtype, and interventions like radiofrequency ablation reshaping the PDAC-ME were also discussed. Additionally, the roles of cancer associated fibroblasts and the therapeutic potential inhibitors in combination with signal transducer and activator of transcription 3 blockade and anti-CD47/anti-PD-L1 immunotherapy were reviewed in preventing PDAC resistance.