Development of Human Intestinal Simulacra

人体肠道模拟物的开发

• 批准号: 8948275
• 负责人: Nancy L. Allbritton
• 金额: $ 90.25万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-25 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8948275
• 关键词: Achievement; Adherent Culture; Architecture; Autistic Disorder; Bacteria; Behavior; Biological Assay; Biological Models; Biology; Biopsy; Butyrates; Caliber; Cardiovascular Diseases; Cell Culture System; Cell Differentiation process; Cell physiology; Cells; Chemicals; Coculture Techniques; Colon; Communities; Controlled Environment; Development; Devices; Diabetes Mellitus; Diet; Dietary Factors; Digestive System Disorders; Dimensions; Disease; Environment; Epithelial; Epithelial Cells; Epithelium; Food; Gases; Genetic Predisposition to Disease; Geometry; Germ-Free; Gnotobiotic; Goals; Growth; Growth Factor; Growth and Development function; Health; Human; Human Development; Hydrogels; Immune Tolerance; In Vitro; Individual; Interdisciplinary Study; Intestinal Diseases; Intestinal Mucosa; Intestines; Investigation; Length; Life; Link; Mediating; Medicine; Metabolic; Metagenomics; Methyl Green; Microbe; Microfabrication; Mitogens; Molecular Conformation; Mucous Membrane; Mus; Organ; Organism; Organoids; Oxygen; Patients; Pattern; Pharmaceutical Preparations; Physiological; Physiology; Porosity; Positioning Attribute; Process; Protocols documentation; Research Personnel; Research Project Grants; Role; Stem cells; Structure; Surface; System; Technology; Testing; Tissues; Tube; Work; advanced system; base; bone morphogenic protein; community living; genome wide association study; gut microbiota; human disease; human stem cells; human tissue; innovation; insight; intestinal epithelium; meetings; metabolomics; microbiome; monolayer; morphogens; mouse model; new technology; novel; prevent; public health relevance; scaffold; self-renewal; stem; stem cell niche; three dimensional structure; tool

 DESCRIPTION (provided by applicant): In the current application, a collaborative, multidisciplinary research project is proposed that has the potential to create a new paradigm for the study of human physiology in health and disease. A state-of-the-art microfabricated platform will be developed to create a functional, in vitro replica, i.e. simulacrum, of the human colonic epithelium and its associated microbiome. This new technology will be used to perform novel studies and hypothesis testing of intestinal physiology that cannot currently be performed. Furthermore, the technology and protocols developed here will establish the basis for creating organ simulacra from normal and diseased primary human tissues that will change the manner in which studies of tissue function, microbiome influence, and drug effect are performed. Recent progress in organotypic culture of colonic epithelial stem cells has made it possible to create long-lived spheroids in a gelatinous matrix. However, the absence of chemical gradients as well as patterned physical support results in a disorganized and chaotic conformation that poorly mimics the structure and function of the colon, and furthermore is not amenable to microbiome co-culture. We propose to develop a novel microfabricated system that will enable ex vivo culture of human colonic epithelium and overlying microbiota that recapitulates the 3D structure and environment of the colonic mucosa in a setting which lies between an unpolarized cell culture system and the complexity of the intact human organism. Recent technical advances from our labs in sustained monolayer culture of colonic epithelial stem cells will be integrated with microfabricated scaffoldings and devices to create the colonic simulacrum. A number of innovations will be incorporated into the microengineered system with the goal of recapitulating the colonic stem-cell niche, the differentiated intestinal mucosa, the microbiota, and the dynamic information flow between these compartments. The platform will permit tight control of the luminal and basal crypt environments by providing independent fluidic and gaseous access to these compartments. The platform will support formation of mitogen, morphogen, differentiation-factor, dietary-compound and gaseous gradients to enable unprecedented investigations into colonic physiology. A number of hypotheses related to the interplay of dietary factors, the microbiome and the colonic epithelium will be tested to demonstrate the power and broad applicability of this transformative technology.

 描述（由申请人提供）：在当前的申请中，提出了一个协作的、多学科的研究项目，该项目有可能为健康和疾病方面的人类生理学研究创造一个新的范式。将开发一个最先进的微加工平台，以创建人类结肠上皮及其相关微生物组的功能性体外复制品，即拟像。这项新技术将用于进行目前无法进行的肠道生理学新研究和假设检验。此外，这里开发的技术和协议将为从正常和患病的原代人体组织创建器官拟像奠定基础，这将改变组织功能、微生物影响和药物作用研究的方式。结肠上皮干细胞器官型培养的最新进展使得在凝胶状基质中产生长寿命的球体成为可能。然而，化学梯度和图案化物理支撑的缺乏会导致无序和混乱的构象，无法很好地模拟结肠的结构和功能，而且不适合微生物组共培养。我们建议开发一种新型微加工系统，该系统将能够对人类结肠上皮和上覆微生物群进行离体培养，从而在介于非极化细胞培养系统和完整人体有机体的复杂性之间的环境中重现结肠粘膜的 3D 结构和环境。我们实验室在结肠上皮干细胞持续单层培养方面的最新技术进展将与微型支架和装置相结合，以创建结肠模拟物。许多创新将被纳入微工程系统中，目的是重现结肠干细胞生态位、分化的肠粘膜、微生物群以及这些区室之间的动态信息流。该平台将通过提供进入这些隔室的独立流体和气体通道来严格控制管腔和基底隐窝环境。该平台将支持有丝分裂原、形态发生素、分化因子、膳食化合物和气体梯度的形成，以便对结肠生理学进行前所未有的研究。将测试与饮食因素、微生物组和结肠上皮相互作用相关的许多假设，以证明这种变革性技术的力量和广泛的适用性。