Engineering of human intestinal organoids containing immune cells

含有免疫细胞的人体肠道类器官工程

• 批准号: 8855062
• 负责人: Christiane Wobus
• 金额: $ 34.95万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-01 至 2020-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8855062
• 关键词: 3-Dimensional; Anti-Infective Agents; Antigen-Antibody Complex; Bacteriology; Behavior; Benchmarking; Cell physiology; Cells; Child; Clostridium difficile; Coculture Techniques; Complex; Crohn' s disease; Development; Diarrhea; Disease; Disease model; Disease susceptibility; Economics; Engineering; Enteral; Enterocytes; Environment; Epithelial; Epithelial Cells; Epithelium; Equilibrium; Functional disorder; Gastrointestinal tract structure; Generations; Goals; Gram-Negative Bacteria; Gram-Positive Bacteria; Hematopoietic; Human; Human Engineering; Immune; Immune response; Immunology; In Vitro; Individual; Infection; Inflammatory Bowel Diseases; Inflammatory Response; Intestinal Diseases; Intestines; Lamina Propria; M cell; Measures; Methods; Modeling; Morbidity - disease rate; Mucosal Immune Responses; Mucosal Immunity; Mutation; Norovirus; Oral; Organ; Organoids; Pathogenesis; Physiological; Pluripotent Stem Cells; Property; Public Health; Research; Research Personnel; Research Project Grants; Rotavirus; Salmonella enterica; Sampling; Smooth Muscle Myocytes; Surface; System; Testing; Therapeutic; Toxin; Vaccination; Vaccines; Villus; Virus; Virus Diseases; Work; cell type; cellular engineering; commensal microbes; cost; disorder control; disorder prevention; drug development; effective therapy; enteric pathogen; food antigen; foodborne illness; gut microbiota; health economics; immune function; improved; in vitro Model; in vivo; innate immune function; intestinal epithelium; intestinal homeostasis; killings; microbial; mortality; multidisciplinary; novel; oral vaccine; pathogen; pathogenic bacteria; response; transcytosis; vaccine efficacy; virology

PROJECT SUMMARY/ABSTRACT Enteric pathogens infect their human host via the gastrointestinal tract causing significant morbidity, mortality, and economic hardship worldwide. The intestine is a complex organ comprised of a layer of epithelial cells, the lamina propria containing immune cells, and a layer of smooth muscle cells organized into villi and crypts. Currently available in vitro models of the human intestine have major limitations, e.g., lack of immune cells or microbiota, and therefore, understanding how human enteric pathogens behave in their native host environment has presented significant challenges. Systems more closely mimicking the human intestine are needed to gain a better understanding of intestinal diseases, host - pathogen interactions and to facilitate the development of more effective therapies. Recent studies revealed that primary human pluripotent stem cells can be differentiated in vitro into three-dimensional human intestinal organoids (HIOs) with a pseudo-lumen and polarized intestinal epithelium similar to the organization found in the intestine in vivo. Importantly, HIOs contain the major secretory and absorptive cell types of the intestinal epithelium. However, HIOs lack microfold (M) cells and immune cells, which are critical for induction of mucosal immunity and balancing of appropriate tolerogenic and/or inflammatory responses to food antigens, commensal bacteria and pathogens. Imbalances in intestinal homeostasis result in devastating diseases such as inflammatory bowel disease (IBD), and M cells are important targets for improving oral vaccine efficacy. Therefore, the goal of Project 2 is to develop immune HIOs containing M cells and immune cells. We hypothesize that immune HIOs, i.e., HIOs co-cultured with immune cells, will more faithfully mimic the human intestine than current models and thus will be superior for studying host-pathogen interactions, drug development and disease pathogenesis. To develop complex human intestinal organoids with immune cells, we will: 1) establish and benchmark a complex organoid model by co-culturing immune cells with HIOs (immune HIOs); 2) determine the response of the complex organoid model to infection with Salmonella enterica serovar Typhimurium, Clostridium difficile and human norovirus; and 3) determine the suitability of immune HIOs with IBD-associated mutations to model aspects of disease pathogenesis. The effects of microbiota-derived products on the function of immune HIOs will also be evaluated. Our studies will synergize with Project 1, which will elucidate the interactions of HIOs with commensal or pathogenic bacteria in the absence of immune cells, and with Project 3, which will engineer a platform for the HIOs to allow intra- and extra-luminal flow and sampling. Our proposed research is significant because it is expected to provide a novel system to facilitate studies of intestinal dysfunction. The immune HIO model has the potential to fast-track development of effective vaccines, anti-infectives and treatment options for intestinal diseases like IBD.

项目总结/摘要 肠道病原体通过胃肠道感染它们的人类宿主，导致显著的发病率、死亡率， 世界范围内的经济困难。肠是由一层上皮细胞组成的复杂器官， 含有免疫细胞的固有层和组织成绒毛和隐窝的平滑肌细胞层。 目前可用的人肠体外模型具有主要局限性，例如，缺乏免疫细胞或 因此，了解人类肠道病原体在其天然宿主中的行为 环境带来了重大挑战。更接近模仿人类肠道的系统是 需要更好地了解肠道疾病，宿主-病原体相互作用， 开发更有效的治疗方法。最近的研究表明，原代人类多能干细胞 可以在体外分化成具有假腔的三维人肠类器官（HIO） 和极化的肠上皮，类似于体内肠中发现的组织。重要的是， 包含肠上皮的主要分泌和吸收细胞类型。然而，HIO缺乏微折叠 (M)细胞和免疫细胞，它们对于诱导粘膜免疫和平衡适当的 对食物抗原、共生细菌和病原体的致耐受性和/或炎症反应。失衡 肠内稳态导致破坏性疾病，如炎症性肠病（IBD）和M细胞 是提高口服疫苗效力的重要靶点。因此，项目2的目标是开发免疫 含有M细胞和免疫细胞的HIO。我们假设免疫HIO，即，共培养的HIO 免疫细胞，将更忠实地模仿人类肠道比目前的模型， 对于研究宿主-病原体相互作用、药物开发和疾病发病机制具有上级优势。到 开发具有免疫细胞的复杂的人类肠道类器官，我们将：1）建立和基准化一个复杂的 通过将免疫细胞与HIO（免疫HIO）共培养来建立类器官模型; 2）确定免疫细胞对类器官模型的应答。 复杂类器官模型感染沙门氏菌血清型鼠伤寒，艰难梭菌和 人诺如病毒;和3）确定具有IBD相关突变的免疫HIO对模型的适用性 疾病的发病机制。微生物源性产品对免疫HIO功能的影响 也将进行评估。我们的研究将与项目1协同，项目1将阐明HIO的相互作用 在没有免疫细胞的情况下与寄生虫或病原菌一起使用，以及与项目3一起使用， 用于HIO的平台，以允许腔内和腔外流动和采样。我们的研究计划是 这是因为它有望提供一种新的系统来促进肠功能障碍的研究。的 免疫HIO模型有可能快速开发有效的疫苗，抗感染药物， 肠道疾病如IBD的治疗选择。