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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

基本信息

批准号：
10651852
负责人：
Jan-Peter van Pijkeren
金额：
$ 41.84万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
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/10651852
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 Etiology Foundations Fructose Glucose Glycolysis Gnotobiotic Goals Growth Health Human Immune Intestines Knowledge Lactic acid Lactobacillus reuteri 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 Viral Virus Work bacterial community base dietary dietary approach drinking water fatty liver disease fitness gut microbiota innovation interleukin-22 knowledge base microbial mutant next generation programs prospective response sugar symbiont therapeutic protein virtual

项目摘要

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，但其潜在机制推动这些变化的动力仍然难以捉摸。除非我们填补了知识库中的这些空白，否则我们将无法进入理解饮食中的糖，益生菌和它们的病毒之间的相互作用的位置;这阻碍了开发合理的方法来使用饮食来提高（工程）益生菌的功效。很长的- 长期目标是（i）揭示饮食和益生菌之间相互作用的机制，他们的病毒，和（ii）开发微生物疗法。本研究计划的目标是（1）阐明- 确定蔗糖促进益生菌肠道共生体乳酸杆菌中噬菌体产生的机制 reuteri，和（2）利用饮食诱导的噬菌体产生来促进定殖和治疗剂的释放。最重要的假设是蔗糖增加噬菌体的产生，从而促进结肠炎。从工程益生菌中释放重组蛋白。拟议工作的理由它的成功完成预计将导致我们对饮食如何影响噬菌体生产，这将开辟新的和令人兴奋的途径来调节肠道微生物群组成，促进益生菌生长，并定制治疗递送。总体假设将通过以下方式进行测试：三个具体目标：（1）表征与噬菌体生产相关的蔗糖代谢途径;（2）确定噬菌体对L. reuteri肠道适应性响应于富含蔗糖的饮食;和（3）使用饮食糖来控制噬菌体介导的裂解和治疗递送。在第一个目标中，靶向诱变是用于剖析蔗糖代谢途径及其产物，以确定蔗糖驱动的 L. reuteri。在第二个目标下，在gnotobiotic小鼠中进行的细菌竞争试验将确定蔗糖驱动噬菌体生产的生态后果;能力降低的同基因突变体代谢蔗糖，并产生噬菌体，有望揭示因果关系。在第三个目标下，L.罗伊氏将被改造为裂解和释放重组白细胞介素-22，以响应特定的糖在脂肪肝疾病的动物模型中，将研究饮食对治疗效果的影响。这研究是创新的，因为一个重要的互惠肠道共生体被用来解开机制，其中蔗糖促进肠道生态系统中的噬菌体生产，这可以应用于开发下一代益生菌这项研究意义重大，因为了解饮食中的糖如何促进噬菌体肠道生态系统的生产，以及生态后果是什么，打开了以前未被探索的改变肠道微生物群组成的机会，这可能包括富集和/或植入益生菌此外，在我们已经证明，膳食糖促进释放重组治疗性蛋白质，为调节肠道中的治疗释放的现实前景奠定了基础使用激活噬菌体介导的裂解的饮食或环境触发物。