Abstract Purpose: Cytokine release syndrome (CRS) after CD19 CAR-T therapy poses a significant risk of morbidity and stands as a formidable obstacle to the success of CD19 CAR-T therapy. Despite the well-established influence of metabolism on immune responses against infectious diseases, its role in CRS during CAR-T therapy is not fully explored. Methods: We performed untargeted LC-MS-based metabolomic profiling on sequential serum samples from 36 acute lymphoblastic leukemia (ALL) patients at distinct post-infusion timepoints: pre-CRS (CRSbefore), CRS peak (CRSₘiddle), and CRS recovery (CRSₐfter). A total of 108 samples were analyzed. Results: Differentially expressed metabolites (DEMs) were firstly investigated across different timepoints. At CRSₘiddle (vs. CRSbefore), 52 metabolites were upregulated and 12 downregulated, predominantly enriched in caffeine metabolism and ascorbate and aldarate metabolism. Post-CRS recovery (CRSₐfter vs. CRSₘiddle) showed 20 upregulated and 55 downregulated metabolites. Upregulated species included amino acids (lysine, L-threonine) and lipids (glycerophosphocholine, Lyso PE). Co-expression clustering revealed metabolites enriched in glycine, serine, leucine, and isoleucine biosynthesis, phenylalanine metabolism, and pantothenate and CoA biosynthesis gradually decreased over time. Comparing CRS grade 1 vs. 2: 15, 44, and 38 DEMs were identified at CRSbefore, CRSₘiddle, and CRSₐfter, respectively. Receiver operating characteristic (ROC) analysis identified six plasma metabolites distinguishing CRS grade 3 from grade 1 patients with strong discriminatory power: upregulated metabolisms (2', 3'-Dideoxy-3'-fluorouridine, PC (24: 0/20: 3 (6, 8, 11) -OH (5), glutaric acid) and downregulated metabolisms (Acetamide, 1-Amino-2-sulfanylethanesulfinic acid, Tyr-Gly-Gly-Trp-Leu). Conclusion: This study provides comprehensive insights into the dynamic remodeling of the plasma metabolome during CD19 CAR-T therapy and reveals metabolic signatures associated with CRS severity in ALL patients. These findings enhance our understanding of metabolic dynamics in CAR-T therapy and may inform strategies for predicting and managing CRS-related complications.
Fu et al. (Mon,) studied this question.