Deciphering the Role of Gut Microbiome in Inflammatory Bowel Disease Using a Canine Patient-Specific Gut-on-a-Chip

项目总结 此NIH Orip K01奖项申请描述了一项为期5年的培训计划，旨在让我获得更多 技能和知识让我可以过渡到一名独立的R01资助的终身教职研究科学家。在……里面 执行拟议的研究和职业发展计划，我将增加我的科学技能并获得 擅长肠道干细胞生物学、微生物群和微流控芯片器官技术。使用这一新的 我将建立一个科学利基，这将使我有别于我的导师，并为 一个强有力的、由外部资助的研究项目。在我的指导团队的支持下，我设计了一个 强大的研究计划，利用我在比较胃肠病学和 分子生物学。具体地说，目标1将展示影响犬传染性法氏囊病的分子和遗传变化。 在开发和验证特定于患者的IBD模型后，该模型可以定量评估 宿主-微生物组串扰的细胞和分子特征。目标2.将允许绘制微生物图谱 利用单细胞响应宿主-微生物组细胞间串扰的签名和上皮完整性 水平多组学(特别是基因组学和转录组学)和RNA原位杂交。与 ORIP的使命声明促进兽医科学家利用他们在比较医学方面的专业知识来 研究人类疾病，我的研究将使我能够运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炎症相关的肠道稳态的变化 肠病。此外，由于Orip支持人类疾病的动物建模，我将使用狗作为 自发动物模型研究肠道微生物区系对肠道上皮细胞的影响 在基因和生理上与人类相似。这项提案产生的结果具有直接的影响 对于人类疾病，因为它们将为启动或 维持肠道的慢性炎症。这些发现可以应用于各种慢性疾病，这些疾病 在流行病学上与微生物群失调和肠道健康紊乱有关(即， 结直肠癌、糖尿病和阿尔茨海默病，仅举几例)。这一知识可以应用于 了解疾病的发展和旨在通过以下途径改变肠道稳态的新疗法 肠道上皮-微生物-免疫轴的紊乱。综上所述，培训目标和职业生涯 此申请中提出的开发活动将促进我成功过渡到独立 研究方向。

PROJECT SUMMARY This NIH ORIP K01 award application describes a 5-year training plan designed to allow me to gain additional skill and knowledge so that I can transition to an independent R01-funded tenure track research scientist. In carrying out the proposed research and career development plan, I will add to my scientific repertoire and acquire expertise in intestinal stem cell biology, microbiome, and microfluidic organ-on-chip technology. Using this newly acquired expertise, I will establish a scientific niche that will set me apart from my mentors and pave the way to a robust, extramurally funded research program. With the support of my mentoring team, I have designed a robust research program that leverages my extensive expertise with comparative gastroenterology and molecular biology. Specifically, Aim 1 will demonstrate molecular and genetic alterations affecting canine IBD following the development and validation of a patient-specific IBD model that can quantitatively assess the cellular and molecular signature of host-microbiome crosstalk. Aim 2. will allow mapping of the microbial signature and epithelial integrity in response to the host-microbiome intercellular crosstalk by utilizing single-cell level multi-omics (especially genomics and transcriptomics) and RNA in situ hybridization. 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 Inflammatory Bowel Disease. Also, as ORIP supports animal modeling of human diseases, I will be using the dog as a spontaneous animal model to investigate the effect of gut microbiota in the intestinal epithelium given their genetic and physiological similarity to humans. The results generated in this proposal have direct implications for human diseases, since they will provide new insights into genetic and transcriptomic alterations initiating or maintaining the chronic inflammation in the gut. Such findings can be applied to various chronic conditions that have been epidemiologically associated with microbiome dysbiosis and disturbances of intestinal health (i.e., Colorectal Cancer, Diabetes Mellitus, and Alzheimer’s Disease, to name a few). This knowledge may be applied to understand disease development and novel therapies aimed at modifying intestinal homeostasis via perturbation of epithelium-microbiome-immune axis in the intestine. In summary, the training goals and career development activities proposed in this application will promote my successful transition into independent research directions.