Decoding Metabolic Symbiosis between Pancreatic Cancer Cells and Cancer-Associated Fibroblasts Using Cultured Tumor Microenvironment

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive cancer with a poor prognosis, largely due to its unique tumor microenvironment (TME) and dense fibrotic stroma. Cancer-associated fibroblasts (CAFs) play a crucial role in promoting tumor growth and metastasis, contributing to the metabolic adaptation of PDAC cells. However, the metabolic interactions between PDAC cells and CAFs are not well-understood. In this study, an in vitro co-culture model was used to investigate these metabolic interactions. Metabolomic analysis was performed under monoculture conditions of Capan−1 PDAC cells and CAF precursor cells, as well as co-culture conditions of PDAC cells and differentiated inflammatory CAF (iCAF). Co-cultured Capan−1 cells displayed significant metabolic changes, such as increased 2-oxoglutaric acid and lauric acid and decreased amino acids. The metabolic profiles of co-cultured Capan−1 and CAFs revealed differences in intracellular metabolites. Analysis of extracellular metabolites in the culture supernatant showed distinct differences between Capan−1 and CAF precursors, with the co-culture supernatant exhibiting the most significant changes. A comparison of the culture supernatants of Capan−1 and CAF precursors revealed different metabolic processes while co-culturing the two cell types demonstrated potential metabolic interactions. In conclusion, this study emphasizes the importance of metabolic interactions between cancer cells and CAFs in tumor progression and highlights the role of TME in metabolic reprogramming.

Exploring the Role of Desmoplastic Physical Stroma in Pancreatic Cancer Progression Using a Three-Dimensional Collagen Matrix Model

“…Moreover, increased proline expression implies that the physical effect and type I collagen can serve as a proline source to maintain PDAC cell proliferation under nutrient-depleted and hypoxic conditions [33,34]. Proline is catalyzed by proline dehydrogenase 1 (PEODH1) to generate glutamine, which is involved in the tricarboxylic acid (TCA) cycle for ATP production [20,35,36]. Furthermore, the notable reduction in tryptophan in Collagen-Capan-1 medium potentially indicates the activation of the tryptophan metabolic pathway in PDAC cells, implying that the mechanical stress of the 3D collagen matrix potentially supports the generation of immune evasion in this 3D PDAC model.…”

“…Briefly, metabolites were extracted by vortexing the culture medium with ice-cold methanol. Subsequently, the metabolites were purified and analyzed by LC-MS according to the specific experimental methodology of a previous study [20].…”

Collagen Lattice Model, Populated with Heterogeneous Cancer-Associated Fibroblasts, Facilitates Advanced Reconstruction of Pancreatic Cancer Microenvironment

“…This highlighted the association between tumor microenvironment maturation and metabolic pathways, particularly pertaining to extracellular metabolite dynamics. Thus, comprehensive metabolic profiling is imperative [40,41]. We subjected the extracellular primary metabolites from the harvested supernatants to rigorous liquid chromatography-mass spectrometry.…”

“…Primary metabolites were analyzed using liquid chromatography-mass spectrometry (LC-MS-8060, Shimadzu, Kyoto, Japan). Specific experimental methods were based on a previous study [41].…”