Deciphering the Pathogenesis of EHEC Infection and the Effects of Bacteria-Based Therapies Using Comparative Gut-on-a-Chip

使用比较芯片肠道破译 EHEC 感染的发病机制和细菌疗法的效果

基本信息

批准号：
10452087
负责人：
Yoko Miyamoto Ambrosini
金额：
$ 18.93万
依托单位：
WASHINGTON STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10452087
关键词：
Acute Advanced Development Alternative Therapies Alzheimer&apos s Disease Animal Model Animal Testing Antibiotic Therapy Bacteria Biochemical Biological Assay Biopsy Canis familiaris Cell Culture Techniques Cells Chronic Clinical Colitis Colonic Diseases Colorectal Cancer Development Diabetes Mellitus Disease Epithelial Epithelial Cells Escherichia coli Escherichia coli EHEC Escherichia coli Infections Experimental Designs Exploratory/Developmental Grant for Diagnostic Cancer Imaging Exposure to Functional disorder Future Gastroenterology Gastrointestinal Injury Genomics Germ-Free Goals Hemolytic-Uremic Syndrome Hemorrhagic colitis Human Immune Immune response In Situ Hybridization In Vitro Individual Infection Intestines Investigation Lead Maps Medicine Microfluidics Mission Modeling Names Neurologic Organ Outcome Oxygen Pathogenesis Pathologic Pathway interactions Patients Peripheral Blood Mononuclear Cell Persons Physiological Physiology Probiotics RNA Research Resolution Resources Rodent Rodent Model Scientist Shiga Toxin Supportive care System Technology Therapeutic Therapeutic Effect Therapeutic Intervention Time United States National Institutes of Health Visualization Work base canine model combinatorial comparative cytotoxic design dysbiosis foodborne gut microbiome high resolution imaging high risk human disease in vitro Model in vivo insight intestinal epithelium intestinal homeostasis metabolomics microbiome microcin multiple omics novel novel therapeutics organ on a chip pathogen physiologic model preclinical development prevent research and development spatiotemporal stem cell biology stem cells transcriptomics

项目摘要

PROJECT SUMMARY This NIH ORIP R21 award application describes a 2-year plan designed to allow us to investigate pathophysiology of Enterohemorrhagic Escherichia coli (EHEC) infection in translational and comparative canine and human in vitro models. Investigation of the gastrointestinal injuries caused by EHEC and the development and assessment of therapeutic interventions have been hampered by the lack of translatable in vitro models that effectively reproduces the typical human colonic disease that progresses to bloody diarrhea and hemolytic uremic syndrome (HUS). The EHEC infection in dogs takes very similar clinical courses to that in humans as canine EHEC infection can naturally lead to mild to severe forms of EHEC infection as well as HUS. In carrying out the proposed research, development of both canine and human EHEC models will be accomplished while utilizing our expertise in veterinary gastroenterology, intestinal stem cell biology, microbiome, and microfluidic organ-on-chip technology. Our proposed study is a high-risk project because canine in vitro modeling of EHEC infection has never been performed before in an organ-chip platform. However, we believe our previous work makes this study highly feasible to develop canine and human EHEC Chips by incorporating colonoids, microbiome, immune cells, EHEC, and bacteria-based treatments into the physiological model. Specifically, Aim 1 will allow development of canine and human EHEC Chips and assessing the contribution of microbiome in the disease pathogenesis while mapping host responses by utilizing single-cell level multi-omics (especially genomics and transcriptomics) and RNA in situ hybridization. Aim 2. will allow assessment of immune contribution in the pathogenesis. Aim 3. will allow further assessment of preventative or therapeutic effects of two well studied bacteria (E.coli Nissle 1917 and MccPDI producing E. coli) on EHEC Chips. Consistent with the ORIP’s mission statements promoting veterinary scientists to employ their expertise in comparative medicine to investigate human diseases, my research will allow me to use my expertise in comparative gastroenterology as well as in primary stem cell culture to investigate alterations in intestinal homeostasis relevant to EHEC infection. The results generated in this proposal have direct implications for in vivo canine EHEC models and ultimately to human EHEC patients, since they will utilize donor-derived colonoids in the experimental designs and provide new insights into transcriptomic alterations in initiating, propagate, or ameliorate the EHEC infection. Our findings on the species similarities and differences in host responses can be applied to various chronic conditions that have been associated with intestinal dysbiosis that occur in both human and dogs (i.e., Colorectal Cancer, Diabetes Mellitus, and Alzheimer’s Disease, to name a few). In summary, the proposed study in this application will allow us to map the host-EHEC crosstalk as well as the effect of bacteria-based therapies by leveraging the single-cell multi-omics analysis, which may lead to a broader impact on uncovering the underlying disease mechanism and developing new preventative or therapeutic anti-EHEC therapy.

项目概要这份 NIH ORIP R21 奖项申请描述了一项为期 2 年的计划，旨在让我们能够进行调查转化犬和比较犬肠出血性大肠杆菌 (EHEC) 感染的病理生理学肠出血性大肠杆菌引起的胃肠道损伤的研究和开发。由于缺乏可转化的体外模型，治疗干预措施和评估受到阻碍有效地再现了典型的人类结肠疾病，该疾病会发展为血性腹泻和溶血狗的肠出血性大肠杆菌感染的临床过程与人类非常相似。犬肠出血性大肠杆菌感染自然会导致轻度至重度的肠出血性大肠杆菌感染以及 HUS。根据拟议的研究，犬和人类肠出血性大肠杆菌模型的开发将在同时完成利用我们在兽医胃肠病学、肠道干细胞生物学、微生物组和微流体方面的专业知识我们提出的研究是一个高风险项目，因为犬肠出血性大肠杆菌的体外建模。以前从未在器官芯片平台上进行过感染，但是我们相信我们之前的工作。使得这项研究通过整合结肠素来开发犬和人类肠出血性大肠杆菌芯片变得非常可行，具体来说，目标是将微生物组、免疫细胞、肠出血性大肠杆菌和基于细菌的治疗方法纳入生理模型中。 1 将允许开发犬类和人类肠出血性大肠杆菌芯片并评估微生物组在疾病发病机制，同时利用单细胞水平多组学（尤其是目标 2. 基因组学和转录组学）和 RNA 原位杂交将允许评估免疫。目标 3. 将允许进一步评估预防或治疗效果。 EHEC 芯片上的两种经过充分研究的细菌（大肠杆菌 Nissle 1917 和产生 McPDI 的大肠杆菌）与 EHEC 芯片一致。 ORIP 的使命宣言促进兽医科学家利用他们在比较医学方面的专业知识研究人类疾病，我的研究将使我能够利用我在比较胃肠病学方面的专业知识以及在原代干细胞培养中研究与肠出血性大肠杆菌感染相关的肠道稳态变化。该提案中产生的结果对体内犬 EHEC 模型有直接影响，并最终对人类肠出血性大肠杆菌患者，因为他们将在实验设计中利用供体来源的结肠，并提供关于转录组改变在引发、传播或改善肠出血性大肠杆菌感染方面的新见解。关于宿主反应的物种相似性和差异的研究可以应用于各种慢性病，与人类和狗的肠道菌群失调有关（即结直肠癌、糖尿病和阿尔茨海默病等）总之，本申请中提出的研究。将使我们能够通过利用单细胞多组学分析，这可能会对发现潜在疾病产生更广泛的影响机制开发和新的预防性或治疗性抗 EHEC 疗法。