Investigating the role of natural and engineered curli fibers in mediating interactions with the gut epithelium

研究天然和工程卷曲纤维在介导与肠道上皮相互作用中的作用

• 批准号: 10174214
• 负责人: Neel Satish Joshi
• 金额: $ 30.51万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10174214
• 关键词: Adhesions; Amyloid fibers; Anti-Inflammatory Agents; Bacteria; Behavior; Biological; Biological Assay; Biological Models; Biological Process; Biological Testing; Biopolymers; Chronic; Collaborations; Communities; Complex; Conflict (Psychology); Development; Disease remission; Encapsulated; Engineered Probiotics; Engineering; Environment; Epithelial; Epithelial Cells; Epithelium; Escherichia coli; Fiber; Foundations; Future; Genes; Genetic Engineering; Genomics; Goals; Height; Homeostasis; Human; Immunohistochemistry; In Vitro; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Institutes; Intestinal Mucosa; Measures; Mediating; Microbe; Microbial Biofilms; Modeling; Molecular; Monitor; Mus; Oral Administration; Organ; Pathogenicity; Patients; Peptide Hydrolases; Peptides; Pharmaceutical Preparations; Physiological Processes; Play; Polymers; Probiotics; Process; Production; Property; Proteins; Role; Sampling; Site; Sodium Dextran Sulfate; System; Techniques; Testing; Therapeutic; Time; Tissue Harvesting; Tissues; Trefoil; Villus; Virulence Factors; Work; base; cellular engineering; commensal bacteria; cytokine; design; extracellular; fitness; gastrointestinal epithelium; gut colonization; host colonization; host-microbe interactions; in vitro Model; in vivo; in vivo Model; inflammatory disease of the intestine; insight; luminescence; microbiome; mouse model; mutant; pathogenic Escherichia coli; residence; response; screening; success

Investigating the role of natural and engineered curli fibers in mediating interactions with the gut epithelium Abstract The importance of the microbiome in maintaining gut function has fueled the study of commensals and probiotics in an attempt to identify strains with therapeutic potential. A common strategy to develop viable therapeutics is then to use the naturally occurring microbe or identify a specific molecular agent that leads to the desired effect and make it into a drug. While this approach has seen some success, it is hindered by a lack of understanding of biological mechanisms, an inability to rationally manipulate organismal fitness in the gut environment, and inadequate delivery mechanisms. We propose an alternative approach to influencing host physiological processes focused on engineering the matrix proteins produced by bacteria during host colonization. Specifically, this proposal will determine the role of naturally occurring and engineered curli fibers in mediating inflammatory processes inside the gut. Curli fibers, which are a proteinaceous component of the E. coli biofilm, have been studied extensively in the context of pathogenic strains because of their ability to mediate adhesion to host tissues and stimulate inflammatory cytokine production. However, recent evidence suggests that the fibers may also play a protective role by increasing barrier function. Interestingly, a probiotic E. coli strain (Nissle) that is commonly used to help maintain remission in inflammatory bowel disease (IBD) patients, is also known to produce copious curli fibers in vitro. We will use genetically engineered strains of Nissle to determine to what extent curli fibers play a role in mediating inflammatory processes in the gut. Simultaneously, we will create Nissle strains that are engineered to display anti-inflammatory cytokines on their curli fibers. The efficacy of these various genetically altered Nissle strains will be measured using a combination of in vitro and in vivo model systems. Notably, we will make use of a Gut-on-a-Chip system developed by our collaborators to gain insight into the molecular mechanisms of curli fiber-epithelium interactions. This system enables the study of higher-order epithelial functions, like barrier function, villus height, and adhesion, much better than conventional transwell assays. Our proposed work with this system will also help validate it as a rapid screening technique for probiotics. In combination with established mouse models of chronic gut inflammation, these model systems will facilitate efficient identification and development of microbes with therapeutic potential against chronic gut inflammation. This is also part of a broader effort in our lab to establish that biofilm matrix proteins can be a versatile new platform for therapeutic delivery and probiotic targeting.

研究天然和工程冰纤维在介导与与的相互作用中的作用 肠上皮 抽象的 微生物组在维持肠道功能方面的重要性加剧了研究 共生和益生菌试图鉴定具有治疗潜力的菌株。一个 然后 确定一种特定的分子剂，导致所需的效果并将其变成药物。尽管 这种方法已经取得了一些成功，因此缺乏对生物学的了解而阻碍 机制，无法理性地操纵肠道环境中的有机体适应性，并且 交付机制不足。我们提出了一种影响主机的替代方法 生理过程的重点是工程于细菌生成的基质蛋白 宿主殖民化。具体而言，该建议将决定自然发生的作用和 在肠道内部介导炎症过程中设计的卷纤维。卷纤维，其中 是大肠杆菌生物膜的蛋白质成分，已在 致病菌株的背景，因为它们能够介导宿主组织的粘附和 刺激炎症性细胞因子产生。但是，最近的证据表明纤维 也可以通过增加屏障功能来发挥保护作用。有趣的是，益生菌大肠杆菌 菌株（Nissle）通常用于帮助维持炎症性肠病的缓解 （IBD）患者也众所周知会在体外产生大量的卷纤维。我们将在基因上使用 nissle的工程菌株以确定卷纤维在何种程度中起作用 肠道中的炎症过程。同时，我们将创建nissle菌株 设计为在其卷纤维上显示抗炎细胞因子。这些的功效 通过体外和IN的组合，将测量各种遗传改变的NISSLE菌株 体内模型系统。值得注意的是，我们将利用由我们的肠道芯片系统开发的 合作者可以深入了解卷纤维上皮相互作用的分子机制。 该系统可以研究高阶上皮功能，例如屏障功能，绒毛 高度和粘附，比传统的Transwell分析要好得多。我们提出的工作 该系统还将有助于将其作为益生菌快速筛查技术验证。结合 借助已建立的慢性肠道炎症的小鼠模型，这些模型系统将有助于 有效鉴定和开发具有治疗潜力的微生物针对慢性 肠炎。这也是我们实验室中更广泛努力的一部分，以确定生物膜矩阵 蛋白质可以成为用于治疗性递送和益生菌靶向的多功能新平台。