A tunable 3D human small intestinal tissue model for study of enteric pathogens

用于研究肠道病原体的可调 3D 人体小肠组织模型

• 批准号: 9222476
• 负责人: Joan C Mecsas
• 金额: $ 24.08万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9222476
• 关键词: 3-Dimensional; Acute; Animal Model; Animals; Anti-Infective Agents; Antibiotics; Bacterial Adhesins; Bile fluid; Biological; Biological Models; Biomedical Engineering; Biomimetics; Cell Line; Cell model; Cells; Chronic; Communicable Diseases; Complex; Cultured Cells; Development; Disease; Enteral; Environment; Epithelial; Epithelial Cells; Epithelium; Event; Food; Genetic Transcription; Goals; HT29 Cells; Human; Immune; Immune response; In Vitro; Infection; Inflammatory; Inflammatory Response; Integration Host Factors; Interruption; Intervention; Intestinal Diseases; Intestines; Lead; Life; Location; Measures; Modeling; Morbidity - disease rate; Mucous Membrane; Myofibroblast; Outcome; Pasteurella pseudotuberculosis; Pathogenesis; Pathogenicity; Physiological; Predisposition; Property; Proteins; Reproducibility; Research Personnel; Rodent Model; Role; Seeds; Silk; Site; Small Intestines; Structure; Study models; System; Testing; Therapeutic; Time; Tissue Model; Tissues; Vibrio cholerae; Villus; Yersinia; base; cell type; cytokine; design; efficacy testing; enteric pathogen; field study; human tissue; in vitro Model; in vivo; intercellular communication; intestinal epithelium; intestinal villi; microbiota; monolayer; mortality; novel; pathogen; physical property; prevent; programs; quorum sensing; response; scaffold; small molecule; three-dimensional modeling; tissue culture; waterborne

ABSTRACT Enteric infections caused by bacterial pathogens are often debilitating and life-threatening. The most common models for studying these pathogens are in vivo rodent models and in vitro intestinal epithelial cell monolayers. However, these models often do not manifest the true outcomes of enteric infections that occurred in the human intestine. Therefore, many aspects of the interactions between these pathogens and the human host remain unknown. This project aims to dissect the intricate host-pathogen interactions for two important intestinal enteric pathogens, Vibrio cholerae and pathogenic Yersinia pseudotuberculosis, using a multicellular 3D in vitro human tissue model that has villi and flow. We will develop and employ a bioengineered model of the human intestine tunica mucosa that mimics the physiological structures and functions of the intestine by introducing primary intestinal cells, flow dynamics, and villus topology to a 3D scaffold. Specifically, we will use silk proteins as scaffolds to develop a 3D multicellular matrix system to support human intestinal epithelium formation for sustained cultivation and for infection by enteric pathogens. This scaffold design is based on our previously developed 3D silk scaffold system seeded with the cultured cell lines Caco-2 and HT-29 cells and primary human myofibroblast cells. Once we incorporate primary epithelial cells, villus-topology and flow dynamics and build this 3D human intestinal model, we will study how V. cholerae and Y. pseudotuberculosis colonize and cause damage to the human intestine. Because these pathogens have very different pathophysiological outcomes on the human intestine they are excellent models to use in exploring the versatility of these novel 3D bio-mimetics of the intestinal system. Our aims are (1) to build and characterize a 3D model human small intestinal tissue with primary cells, flow and villi and (2) investigate the spatial and temporary dynamics of pathogen colonization and damage as well as the pathophysiological responses of the host cells to V. cholerae or Y. pseudotuberculosis in these 3D tissues. The end result will be a robust, tractable, and well-characterized 3D small intestinal tissue model system that can be used by the field for studying the specific mechanistic steps that are important for enteric pathogens to successfully colonize the host intestine. Importantly, these studies will provide a platform upon which to build a larger program in which multiple investigators can use these 3D systems to probe interactions with various enteric pathogens, microbiota, and anti-infectives as well as to further modify these systems to incorporate other cell types and host factors.

抽象的 由细菌病原体引起的肠道感染常常使人衰弱并危及生命。最 研究这些病原体的常见模型是体内啮齿动物模型和体外肠上皮细胞模型 单层。然而，这些模型通常不能体现发生肠道感染的真实结果 在人体肠道中。因此，这些病原体与人类之间相互作用的许多方面 主机仍未知。该项目旨在剖析两个重要的宿主与病原体之间复杂的相互作用 肠道病原体，霍乱弧菌和致病性假结核耶尔森菌，使用多细胞 具有绒毛和流动的 3D 体外人体组织模型。我们将开发并采用生物工程模型 人体肠道粘膜，模仿肠道的生理结构和功能 将原代肠细胞、流动动力学和绒毛拓扑引入 3D 支架。具体来说，我们将使用 丝蛋白作为支架开发 3D 多细胞基质系统来支持人类肠上皮 用于持续培养和肠道病原体感染的形成。这个脚手架的设计是基于我们的 先前开发的 3D 丝支架系统接种了培养细胞系 Caco-2 和 HT-29 细胞， 原代人肌成纤维细胞。一旦我们整合了原代上皮细胞、绒毛拓扑和流动 动力学并建立这个 3D 人体肠道模型，我们将研究霍乱弧菌和假结核杆菌如何 定植并对人体肠道造成损害。因为这些病原体有很大不同 它们是人类肠道的病理生理结果，可用于探索 这些新型肠道系统 3D 生物模拟物的多功能性。我们的目标是 (1) 建立并表征 具有原代细胞、血流和绒毛的人类小肠组织 3D 模型，(2) 研究空间和 病原体定植和损害的暂时动态以及病原体的病理生理反应 这些 3D 组织中的霍乱弧菌或假结核杆菌的宿主细胞。最终结果将是一个稳健的、 易于处理且特征良好的 3D 小肠组织模型系统，可用于现场 研究对于肠道病原体成功定植非常重要的具体机制步骤 宿主肠道。重要的是，这些研究将提供一个平台，在此基础上建立一个更大的项目，其中 多个研究人员可以使用这些 3D 系统来探测与各种肠道病原体的相互作用， 微生物群和抗感染药，以及进一步修改这些系统以纳入其他细胞类型和 宿主因素。