Engineered Probiotics for Closed-Loop Control of Disease-Associated Gut Metabolites in Gut-On-Chip Models

用于闭环控制芯片肠道模型中疾病相关肠道代谢物的工程益生菌

• 批准号: 10572700
• 负责人: Benjamin Michael Woolston
• 金额: $ 15.7万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-15 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10572700
• 关键词: Anti-Inflammatory Agents; Architecture; Biological Assay; Butyrates; Chemicals; Clinical; Complex; Consensus; Consumption; Crohn' s disease; Data; Directed Molecular Evolution; Disease; Electron Transport; Engineered Probiotics; Engineering; Ensure; Environment; Enzymes; Epithelial; Equilibrium; Escherichia coli; Gel; Genetic; Health; Human; Hydrogen Sulfide; In Situ; In Vitro; Individual; Inflammation; Integration Host Factors; Intestines; Kinetics; Link; Logic; Measurement; Metabolic Pathway; Metabolism; Microbe; Mitochondria; Modality; Modeling; Mucous Membrane; Mucous body substance; Organ; Output; Oxygen; Pathway interactions; Patients; Performance; Physiological; Physiology; Play; Probiotics; Production; Property; Publishing; Reactive Oxygen Species; Research; Resolution; Rheology; Role; Structure; Sulfides; System; Technology; Testing; Therapeutic; Thick; Transcriptional Regulation; Tube; Ulcerative Colitis; Validation; Work; base; commensal microbes; design; design-build-test; disorder control; genotoxicity; gut microbiota; high reward; high risk; human tissue; inflammatory marker; interest; mathematical model; microbial; mucosal microbiota; novel; novel therapeutics; operation; oxidation; promoter; response; sensor; small molecule; spatiotemporal; synthetic biology; therapy outcome; transcription factor

PROJECT SUMMARY Engineered commensal microbes represent a promising platform for controlling microbial metabolism in the gut microbiota for therapeutic outcomes. While strains have been successfully engineered to either reduce the concentration of a toxic metabolite or produce a therapeutic one, strains capable of controlling the level of a metabolite within a narrow window have not been developed. Such ‘smart probiotics’, able to dynamically respond to the environment and either produce or consume a compound based on the local concentration, would be particularly useful for stabilizing metabolites which play a concentration-dependent role in host health and disease. For example, ulcerative colitis and Crohn’s disease have been linked to microbially produced hydrogen sulfide (H2S), with a growing consensus that low levels of this molecule have anti-inflammatory properties and support a healthy epithelium, whereas high concentrations of H2S are genotoxic, inhibit mitochondrial function and butyrate oxidation, and potentially weaken the mucosal barrier. Given that H2S concentration varies spatially and temporally throughout the mucosa, controlling H2S within a tight range is not possible with current small- molecule sulfide donors, which release sulfide regardless of local concentration. We propose a new synthetic biology-based approach to controlling microbial metabolites in situ, in which the engineered microbe uses a transcription factor responsive to the metabolite of interest to dynamically balance the expression of metabolic pathways for production and consumption of the metabolite. This will produce a stable, titratable concentration in a manner analogous to a thermostat. In this proposal, we will demonstrate this technology by developing engineered strains of E. coli Nissle to dynamically control the level of H2S in situ, incorporating mathematical modeling and a human organ-chip platform into the design-built-test cycle to achieve robust and stable operation in the complex gut environment. If successful, the proposed research will establish the design rules for a novel synthetic biology control strategy applicable to many gut metabolites with concentration-dependent roles in disease, identify and mitigate host factors that impact engineered strain performance, and facilitate greater translatability of synthetic probiotics.

项目总结