Metabolic dysfunction–associated steatohepatitis exacerbated by Clostridium perfringens–derived ammonia is attenuated by tripeptide DT-109
Pengxiang Qu, Shusi Ding, Yanru Zhang, Yang Zhao, Erfei Song, Liangshuo Hu, Ruike Ding, Wenbin Cao, Yiting Hou, Jia Qi, Juan Zhao, Chenjing Duan, Shuangqing Liu, Chong Shen, Ying Zhao, Yanhong Guo, Zuowen Zheng, Shiwei Luo, Huizhong Hu, Liang Bai, Sihai Zhao, Bo Wang, Shuixiang He, Yi Wu, Xuelian Xiong, Qiutong Wu, Weiwang Gu, Oren Rom, Aimin Xu, Lemin Zheng, Jifeng Zhang, Enqi Liu, Y. Eugene Chen
Pengxiang Qu, Shusi Ding, Yanru Zhang, Yang Zhao, Erfei Song, Liangshuo Hu, Ruike Ding, Wenbin Cao, Yiting Hou, Jia Qi, Juan Zhao, Chenjing Duan, Shuangqing Liu, Chong Shen, Ying Zhao, Yanhong Guo, Zuowen Zheng, Shiwei Luo, Huizhong Hu, Liang Bai, Sihai Zhao, Bo Wang, Shuixiang He, Yi Wu, Xuelian Xiong, Qiutong Wu, Weiwang Gu, Oren Rom, Aimin Xu, Lemin Zheng, Jifeng Zhang, Enqi Liu, Y. Eugene Chen
View: Text | PDF
Research Article Gastroenterology Hepatology

Metabolic dysfunction–associated steatohepatitis exacerbated by Clostridium perfringens–derived ammonia is attenuated by tripeptide DT-109

Abstract

The global prevalence of metabolic dysfunction–associated steatohepatitis (MASH) is rising, driven by a complex interplay of metabolic disturbances, inflammation, and fibrosis, yet effective treatment options remain limited. This study examined the relationships among intestinal microbial dysbiosis, ammonia production, and hepatic CD8+ T cell activity in MASH, then assessed the therapeutic potential of DT-109, a glycine-based tripeptide. We investigated the gut/liver axis across human cohorts and both nonhuman primate and mouse MASH models. Multiomics approaches were used to characterize ileal microbiota, ammonia levels, and hepatic immune and metabolic pathways. Causality was verified through microbiota transplantation, C. perfringens NirA-knockout mutants, and functional validation in vitro and in vivo. The efficacy of DT-109 was evaluated in nonhuman primates and mice. Our results revealed a significant increase in the ammonia-producing gut bacterium C. perfringens, which led to elevated intestinal ammonia and disruption of the intestinal barrier in MASH. Elevated ammonia levels triggered FosB-mediated upregulation of CCL5 in CD8+ T cells, which in turn drove T cell cytotoxicity in the liver. Notably, DT-109 effectively lowered C. perfringens abundance, reduced intestinal ammonia, restored intestinal barrier integrity, and alleviated CD8+ T cell dysregulation in MASH. These results identify a distinct mechanism in which gut-derived ammonia drives CD8+ T cell–mediated MASH and demonstrate that DT-109 effectively targets this axis by inhibiting C. perfringens and reducing ammonia, ultimately ameliorating MASH.

Authors

Pengxiang Qu, Shusi Ding, Yanru Zhang, Yang Zhao, Erfei Song, Liangshuo Hu, Ruike Ding, Wenbin Cao, Yiting Hou, Jia Qi, Juan Zhao, Chenjing Duan, Shuangqing Liu, Chong Shen, Ying Zhao, Yanhong Guo, Zuowen Zheng, Shiwei Luo, Huizhong Hu, Liang Bai, Sihai Zhao, Bo Wang, Shuixiang He, Yi Wu, Xuelian Xiong, Qiutong Wu, Weiwang Gu, Oren Rom, Aimin Xu, Lemin Zheng, Jifeng Zhang, Enqi Liu, Y. Eugene Chen

×

Figure 4

Elevated ammonia-producing gut bacteria in monkeys with MASH.

Options: View larger image (or click on image) Download as PowerPoint
Elevated ammonia-producing gut bacteria in monkeys with MASH. (A) Nonmet...
(A) Nonmetric multidimensional scaling (NMDS) of metagenome among the 3 groups. (B) Heatmap of top 10 differentially expressed bacteria at species level among the 3 groups. Scale bars, normalized species level. (C) Heatmap of top 10 differentially expressed macrogenomic genes among the 3 groups. K00366, referred to as NirA, as indicated by a red box. Scale bars indicate normalized abundance. (D) Comparison of different enriched FAPROTAX (Functional Annotation of Prokaryotic Taxa) functions. Scale bars, normalized functional abundance (*FDR < 0.05) compared with MASH group. The abundance of C. perfringens (E) and NirA (F). Ammonia levels in ileum contents (G), plasma (H), and liver (I). Hepatic ammonia score (J) by Nessler staining. Nessler’s staining (K) (scale bars, 100 μm). (L) Correlations between significantly altered microbiota, MASH-related or intestinal barrier damage parameters, and ammonia concentrations. (M) C. perfringens and NirA in 40 participants from cohort 1. (N) C. perfringens, NirA, and ammonia in ileum contents of mice (n = 4/group). (O) Ammonia between wild-type C. perfringens and C. perfringens ΔNirA mutant strain (n = 6/group). (P) DT-109 within C. perfringens and the control solution was detected (n = 4/group). C. perfringens over 0–12 hours (Q) and ammonia at 12 hours (R) with or without DT-109 (n = 6/group). C. perfringens (S) and NirA (T) with or without DT-109 (n = 6/group). DT-109 (U) and ammonia (V) in mouse intestinal contents after oral gavage of DT-109 (n = 5/time point). Significantly different species (B), genes (C), and functional profiles (D) were analyzed by DESeq2. Data (E–J, M–V) are presented as means ± SEM. Statistical differences were analyzed using Kruskal-Wallis test followed by Dunn’s post hoc test (E and J), 1-way ANOVA followed by Dunnett’s post hoc test (F–I), unpaired t test (O, Q–V), or Mann-Whitney U test (M, N, and P). (L) *FDR < 0.05; **FDR < 0.01. Values above brackets in figures represent P values unless indicated as FDR.