Lipidomic Profiling of Metastatic Lung Cancer-Derived Exosomes Unveiling Ceramides as Potential Biomarkers by Supramolecular Probe Enrichment and Mass Spectrometry Analysis

Tumor-derived exosomes play a crucial role in intercellular communication, significantly influencing cancer metastatic progression. Nonsmall cell lung cancer (NSCLC), the primary cause of cancer-related mortality in solid tumors worldwide, is particularly notorious for its poor prognosis due to distant metastases, especially to the brain and bone. These metastases considerably reduce patient survival and contribute to therapeutic resistance. In this study, we utilized amphiphilic dendrimer-supramolecular-probe-modified nitrocellulose membranes to efficiently enrich exosomes harvested from bone-metastatic, brain-metastatic, and nonmetastatic lung cancer cells. Subsequent comprehensive lipidomic profiling was performed to identify distinct lipid species encapsulated within these exosomes. Two specific ceramide species, Cer d18:1/22:0 and Cer d18:1/24:0, were pinpointed as key lipids associated with the NSCLC metastatic potential. Functional assays demonstrated that exosomes derived from metastatic cells markedly enhanced the migratory capacity of the recipient lung cancer cells. However, the exogenous supplementation of these ceramides could effectively counteract such pro-migratory effects. These findings provide valuable insights into the role of exosome-mediated sphingolipid metabolism in NSCLC metastasis, validate the two ceramides as potential biomarkers, and establish a proof-of-concept strategy for targeting exosomal lipid reprogramming within the metastatic cascade.