Mechanistic studies of Interleukin 10 gene regulation in intestinal CD4 T cells

描述（由申请人提供）：炎症性肠病（IBD），包括克罗恩病和溃疡性结肠炎，是宿主和环境之间复杂相互作用导致肠道微生物群过度炎症反应的结果。尽管存在几种免疫调节疗法，但许多患者仍然承受着严重的疾病负担。由于IBD的发病率在北美和国外都在增加，需要新的治疗途径来更成功地治疗这些疾病。白细胞介素-10是一种二聚体细胞因子，在体内具有有效的抗炎功能。在肠道中尤其如此，IL-10对于防止小鼠和人类过度的肠道炎症至关重要。然而，由于缺乏对IL-10基因调控复杂性的理解，临床上调节IL-10的产生一直受到阻碍。在这里，我们提供了初步证据，证明转录抑制因子Gfi1可以抑制多种CD4+ T细胞谱系中Il10的转录。此外，我们发现在结肠炎的CD45RBhi转移模型中，Gfi1缺陷T细胞不会驱动严重疾病。在这项初步工作的基础上，我们首先提出利用基因和蛋白表达分析、生物信息学和ChIP研究相结合的方法来确定Gfi1调节CD4+ T细胞亚群中Il10的机制。其次，我们将确定对Il10的影响是否是IBD小鼠模型中的保护机制，以及哪些T细胞亚群有助于这种保护。该培训计划由Casey Weaver博士赞助，将允许学习免疫学、遗传学和ibd指导研究方面的各种技术。此外，它将允许科学项目设计，沟通和培训，这将有利于职业生涯作为一个医生-科学家。虽然本项目将通过小鼠模型研究Il10的调控，但我们的分子研究将以IBD患者GWAS的数据为指导。因此，它也应该揭示在人类疾病中的IL10调控。

DESCRIPTION (provided by applicant): Inflammatory Bowel Diseases (IBD), including Crohn's Disease and Ulcerative Colitis, result from complex interactions between host and environment leading to an excessive inflammatory response to the intestinal microbiota. Although several immunomodulatory therapies exist, many patients still experience a severe burden of disease. Due to the fact that the incidence of IBD is increasing both in North America and abroad, new therapeutic avenues are needed to more successfully treat these disorders. Interleukin-10 is a dimeric cytokine with potent anti-inflammatory functions in vivo. This is especially true in the intestinal tract, where IL-10 is essential to prevent excessive enteric inflammation in both mice and humans. However, modulating IL-10 production clinically has been hampered by a lack of understanding of the complexity of IL10 gene regulation. Here, we provide preliminary evidence that the transcriptional repressor Gfi1 acts to repress Il10 transcription in multiple lineages of CD4+ T cells. Furthermore, we show that Gfi1 deficient T cells do not drive severe disease in the CD45RBhi transfer model of colitis. Building on this preliminary work, we first propose to determine the mechanism of Gfi1 regulation of Il10 in CD4+ T cell subsets using a combined approach of gene and protein expression analysis, bioinformatics, and ChIP studies. Secondly, we will identify if the effect on Il10 is the protectiv mechanism in mouse models of IBD, and which T cell subsets contribute to this protection. The training program, sponsored by Dr. Casey Weaver, will allow for the learning of a variety of techniques in immunology, genetics, and IBD-directed research. Additionally, it will allow for scientific project design, communication, and training that will be beneficial for a career as a physician-scientist. While this project will investigate the regulation of Il10 by the use of mous models, our molecular studies will be guided by data from GWAS of humans with IBD. Thus, it should also shed light on IL10 regulation in human disease.