Mechanism and application of sugar-induced phage production by the probiotic gut symbiont Lactobacillus reuteri

益生菌肠道共生菌罗伊氏乳杆菌糖诱导噬菌体产生的机制及应用

• 批准号: 10298981
• 负责人: Jan-Peter van Pijkeren
• 金额: $ 40.78万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-25 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10298981
• 关键词: Acetic Acids; Animal Model; Automobile Driving; Bacteria; Bacteriophages; Biological Assay; Biological Models; Cells; Communities; Consumption; Cytolysis; Development; Diet; Dietary Sucrose; Dietary Sugars; Disaccharides; Ecology; Ecosystem; Engineered Probiotics; Engineering; Engraftment; Enzymes; Ethanol; Etiology; Fatty Liver; Foundations; Fructose; Glucose; Glycolysis; Gnotobiotic; Goals; Growth; Health; Human; Immune; Intestines; Knowledge; Lactic acid; Lactobacillus reuteri; Liver diseases; Maltose; Mannitol; Mediating; Metabolic; Metabolism; Microbe; Modeling; Mus; Mutagenesis; Outcome; Pathway interactions; Positioning Attribute; Probiotics; Production; Publishing; Recombinant Interleukins; Recombinant Proteins; Recombinants; Research; Role; Sucrose; Testing; Therapeutic; Treatment Efficacy; Tweens; Viral; Virus; Work; bacterial community; base; dietary; dietary approach; drinking water; fitness; gut microbiota; innovation; interleukin-22; knowledge base; microbial; mutant; next generation; programs; prospective; response; sugar; symbiont; therapeutic protein; virtual

While dietary sugars can alter the bacteriophage community in the gut ecosystem, the underlying mechanisms driving these changes remain elusive. Until we have filled these voids in our knowledge base, we will not be in a position to comprehend the interplay between dietary sugars, and probiotics and their viruses; this hampers the development of rational approaches to use diet to promote the efficacy of (engineered) probiotics. The long- term goals are (i) to unravel the mechanisms that drive the interplay between diet, and probiotic bacteria and their viruses, and (ii) to develop microbial therapeutics. The objectives of this research program are (1) to eluci- date the mechanisms by which sucrose promotes phage production in the probiotic gut symbiont Lactobacillus reuteri, and (2) to exploit diet-induced phage production to promote colonization and the release of therapeutics. The overarching hypothesis is that sucrose increases phage production, which consequently promotes coloni- zation and the release of recombinant proteins from engineered probiotics. The rationale of the work proposed is that its successful completion is expected to result in a paradigm shift of our understanding how diet impacts phage production, which will open up new and exciting avenues to modulate the gut microbiota composition, promote probiotic growth, and to tailor therapeutic delivery. The overarching hypothesis will be tested by pursuing three specific aims: (1) To characterize the sucrose metabolism pathway in relation to phage production; (2) To determine the role of phage on L. reuteri gut fitness in response to a diet enriched in sucrose; and (3) To use dietary sugar to control phage-mediated lysis and therapeutic delivery. In the first aim, targeted mutagenesis is used to dissect the sucrose metabolism pathway and their products to determine the triggers of sucrose-driven phage production in L. reuteri. Under the second aim, bacterial competition assays in gnotobiotic mice will de- termine the ecological ramifications of sucrose-driven phage production; isogenic mutants with reduced ability to metabolize sucrose, and to produce phage, are expected to reveal causation. Under the third aim, L. reuteri will be engineered to lyse—and release recombinant interleukin-22—in response to metabolism of a specific sugar. In an animal model of fatty liver disease therapeutic efficacy in response to diet will be investigated. This research is innovative because an important mutualistic gut symbiont is used to unravel the mechanisms by which sucrose boosts phage production in the gut ecosystem, which can be applied towards the development of next-generation probiotics. The research is significant because understanding how a dietary sugar boosts phage production in the gut ecosystem, and what the ecological ramifications are, opens up previously unexplored opportunities to alter the composition of the gut microbiota, which may include enrichment and/or engraftment of probiotic bacteria. Also, after we have demonstrated that a dietary sugar promotes the release of a recombinant therapeutic protein, a foundation is created for a realistic prospective to regulate therapeutic release in the gut using dietary or environmental triggers that activate phage-mediated lysis.

虽然饮食中的糖可以改变肠道生态系统中的噬菌体群落，但潜在的机制 推动这些变化仍然难以捉摸。在我们填补知识库中的这些空白之前，我们将不会进入 理解膳食糖、益生菌及其病毒之间的相互作用的立场；这阻碍了 开发合理的方法来使用饮食来促进(工程)益生菌的功效。长的- 学期目标是(I)揭开驱动饮食、益生菌和 他们的病毒，以及(Ii)开发微生物疗法。这项研究计划的目的是(1)阐明- 蔗糖促进益生菌肠道共生菌噬菌体产生的机制 (2)利用食物诱导的噬菌体生产来促进定植和治疗药物的释放。 最重要的假设是蔗糖增加了噬菌体的产量，从而促进了菌落形成。 工程益生菌中重组蛋白的降解和释放。建议开展的工作的基本原理 它的成功完成有望导致我们对饮食影响的理解发生范式转变 噬菌体生产，这将开辟新的令人兴奋的途径来调节肠道微生物区系组成， 促进益生菌生长，并量身定做治疗药物。最重要的假设将通过追查来检验 三个特定的目标：(1)描述与噬菌体生产有关的蔗糖代谢途径；(2) 确定噬菌体在富含蔗糖的饲料中对罗氏乳杆菌肠道适应性的作用；以及(3)使用 控制噬菌体介导的裂解和治疗性递送的饮食糖。在第一个目标中，定向突变是 用于剖析蔗糖代谢途径及其产物，以确定蔗糖驱动的触发因素 路氏乳杆菌噬菌体生产。在第二个目标下，诺生菌小鼠的细菌竞争分析将被取消。 终止蔗糖驱动的噬菌体生产的生态分支；能力降低的同源突变 代谢蔗糖，并产生噬菌体，预计将揭示原因。在第三个目标下，L.reuri 将被改造为裂解和释放重组白介素22，以响应特定的新陈代谢 糖。在脂肪性肝病的动物模型中，将调查饮食反应的治疗效果。这 研究是创新的，因为一种重要的互惠共生体被用来通过 哪种蔗糖可以促进肠道生态系统中噬菌体的产生，这可以用于开发 下一代益生菌。这项研究意义重大，因为了解了饮食中的糖是如何促进噬菌体的 肠道生态系统的生产，以及其生态后果是什么，揭开了以前未曾探索过的 改变肠道微生物区系组成的机会，这可能包括浓缩和/或嫁接 益生菌。此外，在我们已经证明饮食中的糖促进重组人的释放之后 治疗性蛋白质，为现实前景奠定了基础，以调节肠道中的治疗性释放 使用食物或环境触发物激活噬菌体介导的裂解。