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: 10539253
• 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 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10539253
• 关键词: 3-Dimensional; Address; Adverse event; Anaerobic Bacteria; Animal Model; Animals; Antibiotics; Area; Bacillus; Bacteria; Behavior; Biological Assay; Cecum; 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 Cells; Epithelium; Event; Exhibits; Future; Gastrointestinal tract structure; Germination; 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; Proliferating; 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; human old age (65+); 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 cells; technology platform

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 在当前的申请中，提出了一个合作的多学科研究项目， 开发和验证一个创新平台，研究初级人类之间的相互作用 肠细胞和致病微生物艰难梭菌（C. difficile）。一个新的和独特的 工程技术将创造一个芯片上的器官（器官芯片）平台， 维持与管腔非致病性共培养的有活力的人结肠上皮， 致病专性厌氧菌，以研究发生的基本细胞过程， 在人类和微生物细胞之间的相互作用过程中。这项工作将设计，模型和 设计一个三维膀胱细胞培养平台， 粘液层以及模仿粪便物质通过的纯粹力量。的设计 这一系统将使观察和扰动人-微生物细胞-细胞成为可能 相互作用和修改各自的环境，以阐明潜在的细胞和 分子事件是这些相互作用的关键。该提案的一个重要特点是， 实验将利用新的干细胞培养方法， 人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。 所提出的微生理系统产生的实验数据将在未来 提供关于C.预防和治疗方案的关键信息。difficile以及 为研究其他感染性生物体提供了一种新技术， 胃肠道

项目成果

Human 2D Crypt Model for Assaying Intestinal Stem Cell Proliferation and Differentiation.

• DOI: 10.1021/acs.analchem.2c00905
• 发表时间: 2022-07-05
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Wang, Yuli;Sims, Christopher E.;Allbritton, Nancy L.
• 通讯作者: Allbritton, Nancy L.

Suspended Collagen Hydrogels to Replicate Human Colonic Epithelial Cell Interactions with Immune Cells.

• DOI: 10.1002/adbi.202200129
• 发表时间: 2022-11
• 期刊: ADVANCED BIOLOGY
• 影响因子: 3.7
• 作者: Hinman, Samuel S.;Massaro, Angelo;Wang, Yuli;Sims, Christopher E.;Kim, Raehyun;Allbritton, Nancy L.
• 通讯作者: Allbritton, Nancy L.

Development of a Primary Human Intestinal Epithelium Enriched in L-Cells for Assay of GLP-1 Secretion.

• DOI: 10.1021/acs.analchem.2c00912
• 发表时间: 2022-07-12
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Villegas-Novoa, Cecilia;Wang, Yuli;Sims, Christopher E.;Allbritton, Nancy L.
• 通讯作者: Allbritton, Nancy L.