The Function and Microbial Regulation of Innate Lymphoid Cells in IBD

描述(申请人提供)：炎症性肠病(IBD)影响100多万美国人。虽然在许多情况下，药物使我们能够控制疾病，但IBD仍然会导致显著的发病率和死亡率。一条重要的遗传途径 在IBD中发现的是由一种名为白介素23的分子激活的。最近，一种新类型的淋巴细胞被发现，称为先天淋巴样细胞(ILC)，它对IL-23有反应。这项提议的目的是支持我的额外研究培训，以确定功能和 帮助我过渡到独立的研究生涯，专注于IBD免疫细胞的微生物调控。肠道免疫细胞激活的微生物调控专家丹·利特曼博士和微生物依赖的肠道炎症专家蒂莫西·王博士将共同指导这项提案。首先，从IBD(或非IBD)捐赠者的肠道分离出的ILC的遗传物质(即RNA)将使用最先进的测序技术进行定义。我将参加正式的序列分析培训，以便分析这些数据，并确定IBD患者肠道ILC中激活的基因/遗传途径。我建议的第二个主要方面是定义这些肠道ILC与树突状细胞(DC)的相互作用，树突状细胞是一种特殊的免疫细胞，它在炎症部位捕获蛋白质，并将处理后的蛋白质呈递给T细胞，以刺激免疫反应。小鼠肠道中存在几种类型的DC，它们捕获蛋白质和/或刺激T细胞的能力不同。为了研究它们与ILC的相互作用，我将使用我们实验室产生的新型小鼠模型，这些模型允许我用荧光标记跟踪特定的DC亚群，并在活体小鼠中消融特定的DC亚群。此外，我将描述人类结肠中存在的DC亚群，并评估它们在培养中与人类肠道ILCs相互作用的能力。最后，这项工作将使用无菌小鼠来测试来自IBD和非IBD捐赠者的特定细菌支持ILC活动的能力 在结肠炎的小鼠模型中。为了提供科学反馈和职业发展建议，我组建了一个咨询委员会，成员包括IBD免疫学专家劳埃德·梅耶尔博士、人类免疫学专家梅根·赛克斯博士、肠道微生物学专家马丁·布拉泽博士和科研职业发展专家詹姆·鲁宾博士。有了这份科学和职业发展计划，我相信这项建议将为IBD的疾病发病机制以及诊断和治疗策略提供新的见解，并为我过渡到独立的研究生涯奠定坚实的基础。

DESCRIPTION (provided by applicant): Inflammatory bowel disease (IBD) affects over 1 million Americans. Although in many circumstances medications have allowed us to gain control of disease, IBD still results in significant morbidity and mortality. One important genetic pathway identified in IBD is activated by a molecule called interleukin(IL)-23. Recently, a new type of lymphocyte, called an innate lymphoid cell (ILC), was discovered that responds to IL-23. The aim of this proposal is to support my additional research training to characterize the function and regulation of ILCs in IBD and help me transition to an independent research career focused on microbial regulation of immune cells in IBD. Dr. Dan Littman, an expert in the microbial regulation of intestinal immune cell activation, and Dr. Timothy Wang, an expert in microbial-dependent intestinal inflammation, will serve as co-mentors of this proposal. First, genetic material (i.e. RNA) from ILCs isolated from the intestines of patients with IBD (or non- IBD) donors will be defined using state-of-the-art sequencing technology. I will participate in formal training in sequence analysis in order to analyze this data and define genes/genetic pathways activated in intestinal ILCs of patients with IBD. The second main aspect of my proposal is to define the interaction of these intestinal ILCs with dendritic cells (DCs), specialized immune cells that capture proteins at sites of inflammation and present the processed protein to T cells to stimulate an immune response. Several types of DCs exist within the mouse intestine with different capacity to capture proteins and/or stimulate T cells. In order to investigate their interaction with ILCs, I will use novel mouse models generated in our lab that allow me to track particular DC subsets with a fluorescent label and to ablate specific DC subsets in a live mouse. Furthermore, I will characterize the DC subsets present in the human colon and evaluate their ability to interact with human intestinal ILCs in culture. Finally, this work will use "germ-free" mice to test the ability of particular bacteria from IBD and non-IBD donors to support ILC activity in mouse models of colitis. In order to provide scientific feedback and career development advice, I have assembled an Advisory Committee including Dr. Lloyd Mayer, an expert in IBD immunology; Dr. Megan Sykes, an expert in human immunology; Dr. Martin Blaser, an expert in intestinal microbiology; and Dr. Jaime Rubin, an expert in scientific research career development. With this scientific and career development plan, I believe this proposal will offer new insight into disease pathogenesis as well as diagnostic and therapeutic strategies for IBD and serve as a strong foundation for my transition to an independent research career.