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Cell Damage-Like Proteomic Signatures Distinguish Cell-Free and Concentrated Ascites Reinfusion Therapy (CART)
Masaki Nojima1, Goh Murayama1, Moeko Inoue1
1Department of Internal Medicine and Rheumatology, Juntendo University School of Medicine, Tokyo, Japan.
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.
