Wang, Mu‐En

Abstract Introduction: Diffuse large B-cell lymphoma (DLBCL) is the most common type of malignant lymphoma. Previous studies have shown that DLBCL cells are susceptible to GPX4 (Glutathione peroxidase 4)-regulated ferroptosis, an iron-dependent type of programmed cell death characterized by increased reactive oxygen species and excessive lipid peroxidation. In addition to GPX4, FSP1 (Ferroptosis suppressor protein 1, previously known as AIFM2) is a glutathione-independent repressor of ferroptosis that has shown prognostic significance in solid tumors. In this study, we aimed to reveal the significance of GPX4 and FSP1 in DLBCL. Patients and Methods: We performed immunohistochemistry (IHC) for GPX4 and FSP1 in a large cohorts of patients with de novo DLBCL, and evaluated their cytoplasmic and nuclear expression. Prognostic analysis was performed for FSP1 and GPX4 expression in patients treated with rituximab (R)-CHOP or CHOP chemotherapy, respective of TP53 mutation status, as p53 regulates ferroptosis and TP53 mutation is associated with poorer prognosis in DLBCL. Results: FSP1 expression with a ≥10% cutoff was positive in 42.3% of DLBCL cases. Patients with TP53 mutations had a non-significant trend of higher mean FSP1 expression than those with wild-type (Wt) TP53 (P=0.11). Prognostic analysis revealed that cytoplasmic FSP1+ expression was associated with significantly poorer overall survival and progression-free survival in Wt-TP53 DLBCLs treated with R-CHOP (P=0.016 and P=0.003, respectively), and only showed non-significant unfavorable trends in TP53 mutated DLBCLs and patients treated with CHOP. In contrast, GPX4 expression did not show a significant unfavorable prognostic effect in DLBCL, and in fact, was associated with a non-significant trend of better survival. Previously we have quantified immune cell abundance and PD-1/PD-L1 expression in the tumor microenvironment of the study cohort using multiplex fluorescent IHC. Correlative analysis found that FSP1+ patients had significantly higher mean and median abundance of CD68+ cells (both M1 and M2 macrophages) and CD11c+ cells than FSP1- patients. Further analysis in Wt-TP53 and mutated TP53 subcohorts found that only in the TP53 mutated DLBCL subcohort, FSP1 expression was associated with significantly higher mean and median CD163-CD68+ (M1) and overall CD68+ macrophages, whereas in the Wt-TP53 DLBCL subcohort, FSP1 expression was associated with significantly higher median (but not mean) CD163+CD68+ (M2) macrophages and CD11c+ cells. Summary: Cytoplasmic FSP1 expression but not GPX4 had significantly adverse prognostic effect in patients with Wt-TP53 DLBCL treated with standard immunochemotherapy. Our results also suggest that in addition to ferroptosis regulation, macrophage abundance was relevant for the prognostic effects of FSP1 expression. Citation Format: Beibei Lyu, Zijun Xu-Monette, Xiaoxian X. Zhao, Eric D. Hsi, Ming Chen, Carlo Visco, Alexandar Tzankov, Karen Dybkaer, Mu-En Wang, Chang Wang, Qingyan Au, Harry Nunns, Zenggang Pan, Benjamin Parsons, Santiago Montes-Moreno, Fenghuang Zhan, Michael B. Møller, Leon Bernal-Mizrachi, Youli Zu, Shanxiang Zhang, Weina Chen, Govind Bhagat, Yong Li, KEN H. YOUNG. Expression of the ferroptosis suppressor FSP1 but not GPX4 shows significant adverse prognostic effect in diffuse large B-cell lymphoma with wild-type TP53 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 4661.