The Effect of Autophagy on the Generation and Function of Gut Regulatory T Cells

 描述(申请人提供)：炎症性肠病(IBD)是一种慢性胃肠道疾病，是一种复杂的疾病，具有不同的遗传和症状。过去十年的研究已经确定了100多个与IBD发病风险增加有关的基因。全基因组关联研究发现，在一种名为自噬相关16-样1(ATG16L1)的基因上，单核苷酸多态(SNP)与克罗恩病(IBD)的风险增加有关。这种基因在一种称为自噬的过程中发挥作用，这是一种细胞在营养匮乏的情况下回收细胞内成分的过程。这项建议将解决该基因(ATG16L1T300A)的微小变化或多态的影响，并将重点放在一种对免疫系统调节至关重要的细胞类型，称为调节性T细胞(Treg细胞)。由于IBD是一种胃肠道疾病，本研究的重点是这种基因改变如何影响肠道Treg细胞的生成和功能。为了阐明ATG16L1T300A多态在肠道Treg细胞中的作用，我们将采用一种独特的小鼠模型，在该模型中，小鼠与IBD患者具有相同的基因突变。含有荧光Treg细胞的特殊小鼠模型也将被用于帮助在体外鉴定和操纵肠道Treg细胞。在这些小鼠模型中，将使用许多技术和方法来研究肠道Treg细胞，以了解ATG16L1T300A如何影响AIM 1中的Treg细胞功能。使用的一些方法包括：流式细胞术、ELISA、RTqPCR和小鼠结肠炎模型。我们将在AIM 2中使用T细胞上有非常特定受体的额外小鼠品系，例如OT-II T细胞转基因系统。这个系统将使我们能够解决自噬如何影响被称为抗原提呈细胞的免疫细胞处理抗原以识别T细胞的能力。这些细胞非常重要 因为它们指导Treg细胞的发育和功能。通过使用两种结肠炎小鼠模型，并专门观察肠道中的Treg细胞，Aim 3将从机制上理解IBD风险相关基因变化ATG16L1T300A如何在IBD的病理生理学中做出贡献。这项提案产生的数据将为ATG16L1T300A纯合子克罗恩病患者的Treg细胞功能提供新的见解，并将在临床上与IBD精准医学和基于Treg细胞的治疗的进步相关。为此次Ruth L.Kirschstein国家研究服务奖制定的研究培训计划将培养我的沟通和写作技能，并为我成为一名粘膜免疫学领域的独立研究员做好准备。

 DESCRIPTION (provided by applicant): Inflammatory bowel disease (IBD), a chronic disease of the gastrointestinal tract, is a complex disease with heterogeneous underlying genetics and symptoms. Research in the last decade has identified over 100 genes linked to an increased risk for developing IBD. Genome wide association studies have identified a single nucleotide polymorphism (SNP) associated with an increased risk of Crohn's disease, a subset of IBD, in a gene called autophagy-related 16-like 1 (ATG16L1). This gene functions in a process called autophagy, a process by which cells recycle their intracellular components when nutrients are scarce. This proposal will address the effect of a small change, or polymorphism, in this gene (ATG16L1T300A) and will focus on a cell type crucial for immune system regulation, called regulatory T cells (Treg cells). Since IBD is a disease of the gastrointestinal tract, this proposa focuses on how this gene alteration affects the generation and function of gut Treg cells. To elucidate the role of the polymorphism ATG16L1T300A in gut Treg cells, we will employ a unique mouse model in which mice harbor the same genetic mutation as people with IBD. Special mouse models that harbor fluorescent Treg cells will also be used to aid in the identification and manipulation of gut Treg cells in vitro. With these mouse models, gut Treg cells will be studied using many techniques and methods to understand how ATG16L1T300A affects Treg cell function in Aim 1. Some of the methods utilized will include: flow cytometry, ELISA, RTqPCR, and mouse models of colitis. We will use additional mouse strains in Aim 2 that have very specific receptors on their T cells, e.g. the OT-II T cell transgenic system. This system will enable us to address how autophagy affects the ability of immune cells called antigen presenting cells, to process antigen for T cell recognition. These cells are very important as they guide the development and function of Treg cells. By using two mouse models of colitis, and looking specifically at Treg cells in the intestine, Aim 3 will develop a mechanistic understanding of how the IBD-risk associated gene change ATG16L1T300A contributes to the pathophysiology of IBD. Data generated from this proposal will provide novel insight into the function of Treg cells in Crohn's disease patients homozygous for ATG16L1T300A and will be clinically relevant to the advancement of IBD precision medicine and Treg cell-based therapies. The research training plan developed for this Ruth L. Kirschstein National Research Service Award will develop my communication and writing skills and prepare me for a career as an independent researcher in the field of mucosal immunology.