Supplementary file 1_Proteomic analysis of bioprinted pancreatic tumor models under different nutrient conditions.docx

Cancer-associated fibroblasts (CAFs), extracellular matrix components, and tumor cells participate in reciprocal interactions that influence the metabolic state of PDAC tumors.</p>Objective<p>This study investigated nutrient-associated proteomic changes in three-dimensional (3D) bioprinted PDAC–CAF tumor constructs and examined alterations in metabolic and cell-junction-related pathways under nutrient-avid and nutrient-deprived conditions.</p>Methods<p>Three-dimensional bioprinted pancreatic tumor constructs composed primarily of PANC1 cells and cancer-associated fibroblasts (CAFs) were cultured under nutrient-avid (N) and nutrient-deprived (N−) conditions.

Constructs were monitored at Days 0, 6, and 12, and Day 6 was selected for comparative proteomic analysis as an intermediate stage at which nutrient-associated structural divergence was evident while both tumor and stromal compartments remained detectable. Label-free high-resolution LC–MS/MS proteomic profiling was followed by KEGG pathway enrichment and complementary bioinformatic analyses. Selected junction-associated findings were further examined by quantitative real-time PCR.</p>Results<p>Distinct nutrient-associated proteomic signatures were observed in the bioprinted tumor constructs.