Metabolic plasticity enables Clostridioides difficile growth in coculture with neutrophils and other mammalian cells

IF 3.1 3区生物学 Q3 MICROBIOLOGY

Anaerobe Pub Date : 2025-12-01 Epub Date: 2025-10-14 DOI:10.1016/j.anaerobe.2025.103007

Jucy Gabriel , Eleanor Keil , Sanath Chandramouli , Faiza Khondokar , Rachel Wilkinson , Jennifer Trannguyen , Senu Apewokin , Anushri Shankar , Shinsmon Jose

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引用次数: 0

Abstract

Objectives

Clostridioides difficile infection (CDI) is a significant healthcare burden, characterized by severe colitis and high recurrence rates. Although neutrophils migrate rapidly to the colon infected with C. difficile, it does not lead to pathogen clearance. Understanding how pathogen survives within the host remains a knowledge gap. This study investigates host-pathogen interactions in controlled in vitro settings.

Methods

Multiple strains of C. difficile (M7404, VPI10463, R20291, and clinical isolates) were co-cultured with neutrophils, macrophages, and intestinal epithelial cells (IECs) under anaerobic conditions, and bacterial growth was assessed by optical density and micro-replicator-mediated colony assays. Microbial transcriptomics and pathway enrichment analyses were performed 18 h after M7404-neutrophil coculture to understand the metabolic adaptations.

Results

Neutrophils significantly accelerated the growth of C. difficile in anaerobic co-culture. Transcriptomic analysis revealed extensive metabolic reprogramming in C. difficile, including upregulation of oxidative phosphorylation, carbon fixation, and fermentation pathways, suggesting that the bacterium exploits host-derived nutrients to its advantage. Further highlighting the microbial metabolic switch towards utilizing host-derived nutrients, we observed the upregulation of glycerol kinase and the ethanolamine utilization protein. The downregulation of TCA cycle enzymes and upregulation of catalases suggested a shift away from oxidative metabolism and an effort to mitigate neutrophil-induced stress. C. difficile bacterium underwent similar transcriptional programming in coculture with macrophages and IECs.

Conclusions

The transcriptional regulation of C. difficile metabolic genes in the presence of mammalian cells reveals a key virulence trait that enables pathogen persistence within a host. This study also highlights a paradoxical role of neutrophils in CDI, where it's presence may inadvertently enhance pathogen survival. Targeting these metabolic interactions could lead to novel therapeutic strategies for mitigating CDI severity.

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代谢可塑性使艰难梭菌在与中性粒细胞和其他哺乳动物细胞共培养中生长。

目的：艰难梭菌感染（CDI）是一个重要的医疗负担，其特点是严重的结肠炎和高复发率。虽然中性粒细胞迅速迁移到感染艰难梭菌的结肠，但它不会导致病原体清除。了解病原体如何在宿主体内存活仍然是一个知识缺口。本研究在体外控制条件下研究宿主-病原体相互作用。方法：将多株艰难梭菌（M7404、VPI10463、R20291和临床分离株）与中性粒细胞、巨噬细胞和肠上皮细胞（IECs）在厌氧条件下共培养，通过光密度和微复制因子介导的集落试验评估细菌生长情况。在m7404 -中性粒细胞共培养18小时后进行微生物转录组学和途径富集分析，以了解代谢适应。结果：中性粒细胞显著促进了厌氧共培养中艰难梭菌的生长。转录组学分析揭示了艰难梭菌广泛的代谢重编程，包括氧化磷酸化、碳固定和发酵途径的上调，这表明细菌利用宿主来源的营养物质来发挥其优势。进一步强调微生物代谢转向利用宿主来源的营养物质，我们观察到甘油激酶和乙醇胺利用蛋白的上调。TCA循环酶的下调和过氧化氢酶的上调表明从氧化代谢的转变和减轻中性粒细胞诱导的应激的努力。艰难梭菌在与巨噬细胞和内皮细胞共培养时也经历了类似的转录编程。结论：哺乳动物细胞存在时艰难梭菌代谢基因的转录调控揭示了使病原体在宿主内持续存在的关键毒力特性。这项研究还强调了中性粒细胞在CDI中的矛盾作用，它的存在可能会无意中提高病原体的存活率。针对这些代谢相互作用可能会导致新的治疗策略来减轻CDI的严重程度。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Anaerobe 生物-微生物学

CiteScore

5.20

自引率

8.70%

发文量

137

审稿时长

76 days

期刊介绍： Anaerobe is essential reading for those who wish to remain at the forefront of discoveries relating to life processes of strictly anaerobes. The journal is multi-disciplinary, and provides a unique forum for those investigating anaerobic organisms that cause infections in humans and animals, as well as anaerobes that play roles in microbiomes or environmental processes. Anaerobe publishes reviews, mini reviews, original research articles, notes and case reports. Relevant topics fall into the broad categories of anaerobes in human and animal diseases, anaerobes in the microbiome, anaerobes in the environment, diagnosis of anaerobes in clinical microbiology laboratories, molecular biology, genetics, pathogenesis, toxins and antibiotic susceptibility of anaerobic bacteria.