A fungi-derived cyclic peptide enhances Th9-mediated antitumor immunity by targeting ZAP70 and SREBP1
Wenli Zhao, Yang Zhou, Yuyang Chen, Yicheng Sun, Jiaxin Tang, Yihan Zhu, Jie Ren, Tianxu Du, Handuo Wang, Yuan Gao, Yu Hu, Ling Jiang, Tomohiko Ohwada, Qi Luo, Enguang Bi
Wenli Zhao, Yang Zhou, Yuyang Chen, Yicheng Sun, Jiaxin Tang, Yihan Zhu, Jie Ren, Tianxu Du, Handuo Wang, Yuan Gao, Yu Hu, Ling Jiang, Tomohiko Ohwada, Qi Luo, Enguang Bi
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Research Article Immunology Oncology

A fungi-derived cyclic peptide enhances Th9-mediated antitumor immunity by targeting ZAP70 and SREBP1

Abstract

Adoptive cell therapy (ACT) relies on durable and functional T cells to mediate tumor clearance. Th9 cells are a metabolically fit CD4+ T cell subset with strong persistence but limited cytotoxicity. Here, we identified endomelipeptide A (EpA), a cyclic peptide isolated from Ganoderma lucidum–associated endophytic fungi, as a potent enhancer of Th9 cell differentiation. EpA promoted a cytotoxic Th9 phenotype with enhanced mitochondrial function and metabolic fitness. Mechanistically, EpA dually targeted ZAP70 and SREBP1, coupling T cell receptor signaling activation with lipid metabolism suppression. EpA-treated Th9 cells mediated robust, CD8+ T cell–dependent tumor control and enhanced the efficacy of human Th9 CAR T cell therapy in vivo. These findings establish EpA as a distinct cyclic peptide that reprograms Th9 cells and provides a potential approach to boost ACT efficacy.

Authors

Wenli Zhao, Yang Zhou, Yuyang Chen, Yicheng Sun, Jiaxin Tang, Yihan Zhu, Jie Ren, Tianxu Du, Handuo Wang, Yuan Gao, Yu Hu, Ling Jiang, Tomohiko Ohwada, Qi Luo, Enguang Bi

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Figure 8

EpA promotes human Th9 cell differentiation and enhances the antitumor activity of Th9 CAR T cells.

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EpA promotes human Th9 cell differentiation and enhances the antitumor a...
(A and B) Flow-cytometric analysis of IL-9 production in human CD4+ T cells cultured under Th9-polarizing conditions with or without EpA treatment during in vitro differentiation. Representative plots (A) and quantification (B) (n = 4). (C) Quantitative PCR analysis of IL9, IRF4, and SPI1 mRNA expression in human Th9 cells (n = 3). (D) Flow-cytometric analysis of mitochondrial membrane potential in human Th9 cells (n = 3). (E) Luciferase-based assay analysis of intracellular ATP levels in human Th9 cells (n = 5). (F) Flow-cytometric analysis of phosphorylated ZAP70 in human Th9 cells (n = 3). (G) Flow-cytometric analysis of TNF-α expression in human Th9 cells (n = 4). (H) Cytotoxicity assay of EpA-treated or control CD19 CAR Th9 cells against Raji lymphoma target cells at indicated effector-to-target (E/T) ratios (n = 3). (I) Schematic diagram of the in vivo Raji-luciferase xenograft model in NSG mice receiving control or EpA-treated human Th9 CAR T cells. (J and K) Representative bioluminescence imaging (J) and quantification of tumor radiance (K) in NSG mice at indicated time points after adoptive transfer. (L) Kaplan-Meier survival curves of NSG mice treated as in J. Data in J–L are shown from n = 5 mice per group. (M) Kaplan-Meier survival analysis of patients with BRCA, SARC, and LIHC from the TCGA dataset based on the expression of the top 10 most upregulated genes (Il9, H19, Fbxl21, Myh11, Lhx6, Rab27b, C1s1, Hao1, Ccdc159, Il1rn) in EpA-Th9 cells. Statistical significance was determined using 2-tailed unpaired Student’s t test (B–H), 2-way ANOVA followed by Šidák’s post hoc test (K), and log-rank test (L). Data are presented as mean ± SEM. *P < 0.05, ***P < 0.001, ****P < 0.0001. Schematic in I created using BioRender (https://Biorender.com).