Development of colonic intraepithelial lymphocytes and their role in epithelial and immune function

结肠上皮内淋巴细胞的发育及其在上皮和免疫功能中的作用

• 批准号: 9889102
• 负责人: Kristine A. Kuhn
• 金额: $ 12.96万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9889102
• 关键词: 16S ribosomal RNA sequencing; Affect; Anatomy; Antibiotics; Antigens; Bacteria; Basic Science; Bioinformatics; Blood Circulation; CD4 Positive T Lymphocytes; CD8B1 gene; Clinical Research; Coculture Techniques; Colitis; Colon; Colonoscopy; Data; Development; Disease; Distal; Doctor of Philosophy; Epithelial; Epithelial Cells; Epithelium; Etiology; Fluorescence; Foundations; Gene Deletion; Goals; Health; Homeostasis; Human; Immunologics; In Vitro; Inflammation; Inflammatory Bowel Diseases; Interleukin-6; Intestines; Knowledge; Label; Lead; Learning; Light; Link; Lymphocyte; Lymphocyte Function; MYD88 deficiency; Mentors; Mentorship; Methods; Modeling; Molecular Biology; Molecular and Cellular Biology; Monitor; Mucous Membrane; Mus; Oral; Ovalbumin; Pathogenesis; Pathology; Pathway interactions; Patients; Permeability; Population; Positioning Attribute; Production; Property; Research; Role; Sampling; Signal Transduction; Site; Small Intestines; T-Cell Receptor; T-Lymphocyte; Testing; Therapeutic; Tissues; Training; Transgenic Mice; Translating; Translational Research; Viola; career; career development; clinical application; commensal bacteria; commensal microbes; cytokine; dysbiosis; experience; experimental study; host microbiota; immune function; in vitro Model; in vivo; interest; intraepithelial; microbiome; microbiome analysis; microbiome research; microbiome sequencing; mouse model; novel; population based; public health relevance; recruit; response; rheumatologist; targeted treatment; trafficking; translational study

 DESCRIPTION (provided by applicant): As a rheumatologist, I am interested in understanding the etiology of extra-intestinal manifestations of inflammatory bowel disease (IBD). Treatment of patients with extra-intestinal IBD is difficult due to few therapeutic options. My ultimate career goal is to understand the mechanism by which extra-intestinal manifestations arise so that targeted therapies may be developed. In this proposal I hypothesize that resident bacteria educate the colonic intraepithelial lymphocyte (IEL) population, which in turn, influences downstream epithelial and immune functions that result in disease. We chose to focus upon colonic IELs in this proposal so that we can begin to link microbiome studies in human populations that are based on colonic samples to downstream mucosal and immune functions. In order to complete these studies and accomplish my ultimate goal of translating my basic research into clinical applications, I require additional career training that includes in vitro modeling, molecular biology, bioinformatics, and translational science. In Aim 1, we will define the role of commensal bacteria in developing the colonic IEL repertoire. Our preliminary data in mice demonstrate that activated, IL-6+ IELs reside in the colon during homeostasis but are depleted when mice are placed on broad- spectrum antibiotics. For this aim, first I will learn microbiome sequencing and analysis through courses in bioinformatics and my mentorship team. Second, we will utilize ovalbumin-specific T cell receptor transgenic mice that will be given oral ovalbumin to determine if T cell antigen recognition alone is sufficient to establish th colonic IEL population. Third, we will utilize conditional gene deletions of MyD88 in IELs and epithelial cells to better understand innate signals that lead to IEL recruitment into the colonic epithelium. Aim 2 will identify the mechanisms by which IEL secreted IL-6 contributes to barrier function through the use of IL-6-/- mice and in vitro model epithelia. I will learn epithelial biolgy in these experiments from my primary mentor Sean Colgan, PhD who has vast expertise in this field. Finally, Aim 3 will focus on the role of trafficking IELs. We will utilize a novel transgeni mouse wherein colonoscopy-induced green-to-red fluorescence is converted, labeling IELs in the distal colon. Again, Dr. Colgan's knowledge of molecular and cellular biology will guide these experiments. In combination with antibiotic-induced dysbiosis, IL-6 deficiency, and a spontaneous model of colitis, we will be strongly positioned to dissect the impacts of resident intestinal bacteria, IL-6, and inflammation on extra- intestinal IEL trafficking. By understanding the development and function of IELs in health and disease, we hope to identify a target pathway for treatment of extra-intestinal IBD. The project described in this proposal in addition to the career development activities will allow me translate our findings into clinical studies correlating microbiome data, IELs, and disease status in patients with IBD and extra-intestinal IBD. Such translational studies will form the foundation of my independent research career.

 描述（由申请人提供）：作为一名风湿病学家，我有兴趣了解炎症性肠病（IBD）肠外表现的病因学。由于治疗选择很少，肠外 IBD 患者的治疗很困难。我的最终职业目标是了解肠外表现产生的机制，以便开发靶向治疗。在该提案中，我假设常驻细菌会培养结肠上皮内淋巴细胞 (IEL) 群体，进而影响下游上皮和免疫功能，从而导致疾病。我们选择在本提案中重点关注结肠 IEL，以便我们可以开始将基于结肠样本的人群微生物组研究与下游粘膜和免疫功能联系起来。为了完成这些研究并实现将基础研究转化为临床应用的最终目标，我需要额外的职业培训，包括体外建模、分子生物学、生物信息学和转化科学。在目标 1 中，我们将定义共生细菌在发展结肠 ​​IEL 库中的作用。我们在小鼠身上的初步数据表明，在体内平衡期间，活化的 IL-6+ IEL 驻留在结肠中，但当小鼠接受广谱抗生素治疗时，IL-6+ IEL 会被耗尽。为了这个目标，首先我将通过生物信息学课程和我的导师团队学习微生物组测序和分析。其次，我们将利用口服卵清蛋白的卵清蛋白特异性 T 细胞受体转基因小鼠来确定仅 T 细胞抗原识别是否足以建立结肠 IEL 群体。第三，我们将利用 IEL 和上皮细胞中 MyD88 的条件基因删除来更好地了解导致 IEL 募集到结肠上皮的先天信号。目标 2 将通过使用 IL-6-/- 小鼠和体外模型上皮来确定 IEL 分泌的 IL-6 促进屏障功能的机制。我将在这些实验中向我的主要导师 Sean Colgan 博士学习上皮生物学，他在该领域拥有丰富的专业知识。最后，目标 3 将重点关注贩运 IEL 的作用。我们将利用一种新型转基因小鼠，其中结肠镜检查诱导的绿色荧光转变为红色荧光，标记远端结肠中的 IEL。科尔根博士的分子和细胞生物学知识将再次指导这些实验。结合抗生素引起的生态失调、IL-6 缺乏和自发性结肠炎模型，我们将有能力剖析肠道常驻细菌、IL-6 和炎症对肠外 IEL 运输的影响。通过了解 IEL 在健康和疾病中的发展和功能，我们希望找到治疗肠外 IBD 的目标途径。除了职业发展活动之外，本提案中描述的项目将使我能够将我们的发现转化为与 IBD 和肠外 IBD 患者的微生物组数据、IEL 和疾病状态相关的临床研究。这种转化研究将构成我独立研究生涯的基础。

项目成果

Microbiota-mediated mucosal inflammation in arthritis.

微生物群介导的关节炎中粘膜炎症。

• DOI: 10.1016/j.berh.2020.101492
• 发表时间: 2019-12
• 期刊: Best practice & research. Clinical rheumatology
• 作者: Chriswell ME;Kuhn KA
• 通讯作者: Kuhn KA

The arthritis connection to inflammatory bowel disease (IBD): why has it taken so long to understand it?

• DOI: 10.1136/rmdopen-2020-001558
• 发表时间: 2021-04
• 期刊: RMD open
• 影响因子: 6.2
• 作者: Ashrafi M;Kuhn KA;Weisman MH
• 通讯作者: Weisman MH