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

描述(申请人提供)：炎症性肠病(IBD)，包括克罗恩病和溃疡性结肠炎，是由于宿主和环境之间的复杂相互作用导致对肠道微生物区系的过度炎症反应所致。虽然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在北美和国外的发病率都在增加，因此需要新的治疗途径来更成功地治疗这些疾病。白介素10是一种在体内具有强大抗炎功能的二聚体细胞因子。在肠道中尤其如此，IL-10对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基因调控的复杂性的了解，临床上调节IL-10的产生一直受到阻碍。在这里，我们提供了转录抑制因子Gfi1在多个CD4T细胞系中抑制IL10转录的初步证据。此外，我们还发现，在结肠炎的CD45RBi转移模型中，缺乏Gfi1的T细胞并不会导致严重的疾病。在这项前期工作的基础上，我们首次提出采用基因和蛋白质表达分析、生物信息学和芯片研究相结合的方法来确定Gfi1对CD4T细胞亚群中IL10的调节机制。其次，我们将确定在IBD小鼠模型中，对IL10的影响是否为保护机制，以及哪些T细胞亚群参与了这种保护。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指导的研究中的各种技术。此外，它将允许科学的项目设计、交流和培训，这将有利于作为一名内科科学家的职业生涯。虽然这个项目将通过使用MOUS模型来研究IL10的调节，但我们的分子研究将以患有IBD的人类GWAs的数据为指导。因此，这也应该有助于揭示IL10在人类疾病中的调控作用。

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.