Cell Damage-Like Proteomic Signatures Distinguish Cell-Free and Concentrated Ascites Reinfusion Therapy (CART) Membranes in Malignant Ascites: A Mechanistic Study.

Abstract

Introduction: Cell-free and concentrated ascites reinfusion therapy (CART) alleviates the symptoms of malignant ascites but may trigger inflammation. We investigated why AHF-MO filter (A-A)-processed concentrates showed weaker bioactivity than EC20W filter (A-E)-processed concentrates.

Methods: Ascites from three patients with ovarian cancer were processed into untreated ascites (UA), A-A, or A-E. Whole ascites, extracellular vesicles (EVs), and EV-depleted supernatants were injected intraperitoneally into mice. Serum cytokines were measured, and label-free proteomics was used to identify inflammatory pathways.

Results: A-E induced the strongest proinflammatory and Th17/type I interferon responses compared to UA. A-A produced intermediate activation. EV-depleted A-E supernatants remained highly bioactive. A-E was enriched in proteins associated with cellular damage pathways and proinflammatory signaling. A-A exhibited attenuated changes and preserved regulatory proteins.

Conclusion: A-A yields a more balanced inflammatory and proteomic profile than A-E. Our findings provide a preliminary mechanistic rationale for its use in CART and offer targets for membrane optimization.