Development of a microphysiologic system to assay the interaction of the human colonic epithelium on Clostridium difficile

开发微生理系统来测定人结肠上皮对艰难梭菌的相互作用

• 批准号: 10321276
• 负责人: Nancy L. Allbritton
• 金额: $ 54.01万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10321276
• 关键词: 3-Dimensional; Address; Adverse event; Age; Anaerobic Bacteria; Animal Model; Animals; Antibiotics; Area; Bacillus; Bacteria; Behavior; Biological Assay; Cell Communication; Cell Compartmentation; Cell Culture Techniques; Cell Differentiation process; Cell physiology; Cells; Centers for Disease Control and Prevention (U.S.); Cessation of life; Clindamycin; Clinical; Clostridium difficile; Coculture Techniques; Colon; Data; Development; Diarrhea; Disease; Engineering; Environment; Epithelial; Epithelial Cells; Event; Exhibits; Future; Gastrointestinal tract structure; Goals; Growth; Hamsters; Human; Human Microbiome; Immune response; In Vitro; Incidence; Individual; Infection; Infectious Agent; Interdisciplinary Study; Intestines; Invaded; Investigation; Large Intestine; Mammalian Cell; Mediating; Methods; Microbe; Modeling; Molecular; Morbidity - disease rate; Mucous body substance; Mus; Organoids; Pathogenicity; Patients; Perforation; Physiological; Prevention; Process; Production; Race; Recurrence; Reproduction spores; Research Project Grants; Resistance; Risk; Sampling; Screening procedure; Sepsis; Source; System; Techniques; Technology; Testing; Time; Tissues; Toxic Megacolon; Toxin; Tumor Cell Line; Vancomycin; Work; base; cell type; clinically relevant; cost estimate; design; diarrheal disease; experimental study; fecal transplantation; health care settings; human adult stem cell; human disease; human model; improved; innovation; interest; intestinal epithelium; matrigel; microbial; microbiome; microbiome alteration; microbiota; microphysiology system; mortality; novel; organ on a chip; pathogen; pathogenic microbe; response; sex; stem cell proliferation; stem cells

Project Summary In the current application, a collaborative, multidisciplinary research project is proposed to develop and validate an innovative platform to study the interactions between primary human intestinal cells and the pathogenic microbe, Clostridium difficile (C. difficile). A new and uniquely engineered technology will create an intestine-on-chip (organ-on-chip) platform capable of maintaining a viable human colonic epithelium co-cultured with luminal nonpathogenic and pathogenic obligate anaerobes in order to study fundamental cellular processes that occur during the interaction between the human and microbial cells. The work will design, model and engineer a three-dimensional intestinal-cell culture platform that supports a physiologic mucous layer as well as mimics the sheer forces of the passage of fecal material. The design of this system will make it possible to observe and perturb the human-microbe cell-cell interactions and modify respective environments to elucidate the underlying cellular and molecular events key to these interactions. A powerful attribute of the proposal is that these experiments will leverage new stem-cell culture methods that produce a normal, primary human colonic epithelium rather than use tumor cell lines as the source of the mammalian cells. The experimental data generated by the proposed microphysiological system will in the future provide key information concerning prevention and treatment option(s) for C. difficile as well as provide a new technique for the study of other infectious organisms involving the gastrointestinal tract.

项目摘要 在当前的应用程序中，提出了一个合作的多学科研究项目 开发和验证创新平台来研究主要人类的相互作用 肠细胞和致病性微生物，艰难梭菌（艰难梭菌）。一个新的独特 工程技术将创建一个能够的肠道（芯片器官）平台 维持可行的人类结肠上皮，与腔内非对病原体共同培养 致病性障碍厌氧菌，以研究发生的基本细胞过程 在人与微生物细胞之间的相互作用中。工作将设计，模型和 工程师一个支持生理学的三维肠道培养平台 粘液层以及模仿粪便材料通过的纯粹力量。设计 该系统将有可能观察和扰动人类微型细胞细胞 相互作用并修改各自的环境，以阐明潜在的细胞和 分子事件是这些相互作用的关键。该提案的有力属性是这些 实验将利用新的干细胞培养方法，产生正常的原发性 人类结肠上皮而不是使用肿瘤细胞系作为哺乳动物细胞的来源。 拟议的微生物生理系统生成的实验数据将来会 为艰难梭菌以及 为研究其他传染性生物提供了一种新技术，涉及 胃肠道。