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

 描述（由申请人提供）：炎症性肠病（IBD）是一种慢性胃肠道疾病，是一种具有异质性潜在遗传学和症状的复杂疾病。过去十年的研究已经确定了100多个与IBD风险增加有关的基因。全基因组关联研究已经确定了与克罗恩病（IBD的一个子集）风险增加相关的单核苷酸多态性（SNP），该基因称为自噬相关16-like 1（ATG 16 L1）。这种基因在一个称为自噬的过程中发挥作用，这是一个细胞在营养缺乏时回收细胞内成分的过程。该提案将解决该基因（ATG 16 L1 T300 A）中的微小变化或多态性的影响，并将重点关注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，因此该提案重点关注这种基因改变如何影响肠道Treg细胞的生成和功能。为了阐明多态性ATG 16 L1 T300 A在肠道Treg细胞中的作用，我们将采用一种独特的小鼠模型，其中小鼠具有与IBD患者相同的基因突变。携带荧光Treg细胞的特殊小鼠模型也将用于帮助体外鉴定和操作肠道Treg细胞。通过这些小鼠模型，将使用许多技术和方法研究肠道Treg细胞，以了解ATG 16 L1 T300 A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和结肠炎小鼠模型。我们将在Aim 2中使用在其T细胞上具有非常特异性受体的其他小鼠品系，例如OT-II T细胞转基因系统。该系统将使我们能够解决自噬如何影响称为抗原呈递细胞的免疫细胞的能力，以处理抗原用于T细胞识别。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，并特别观察肠道中的Treg细胞，Aim 3将对IBD风险相关基因变化ATG 16 L1 T300 A如何促进IBD的病理生理学产生机制性理解。该提案产生的数据将为ATG 16 L1 T300 A纯合子克罗恩病患者中Treg细胞的功能提供新的见解，并将与IBD精准医学和Treg细胞治疗的进展具有临床相关性。研究培训计划制定了这个露丝L。Kirschstein国家研究服务奖将培养我的沟通和写作能力，并为我作为粘膜免疫学领域的独立研究人员做好准备。

 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.