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，由宿主和环境之间的复杂相互作用引起，导致对肠道微生物群的过度炎症反应。尽管存在几种免疫调节疗法，但许多患者仍然经历严重的疾病负担。由于IBD的发病率在北美和国外都在增加，因此需要新的治疗途径来更成功地治疗这些疾病。 白细胞介素-10是一种二聚体细胞因子，在体内具有强效抗炎作用。在肠道中尤其如此，其中IL-10对于防止小鼠和人类的过度肠道炎症至关重要。然而，由于对IL-10基因调控的复杂性缺乏了解，在临床上调节IL-10的产生受到阻碍。在这里，我们提供了初步的证据表明，转录抑制因子Gfi 1的作用，以抑制IL 10在多个谱系的CD 4 + T细胞的转录。此外，我们发现Gfi 1缺陷型T细胞在结肠炎的CD 45 RBhi转移模型中不会导致严重疾病。 在此基础上，我们首先提出了一种结合基因和蛋白表达分析、生物信息学和ChIP研究的方法来确定Gfi 1调控IL 10在CD 4 + T细胞亚群中的机制。其次，我们将确定对IL 10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。 由凯西韦弗博士赞助的培训计划将允许学习免疫学，遗传学和IBD指导研究的各种技术。此外，它将允许科学项目设计，沟通和培训，这将有利于作为一个医生科学家的职业生涯。 虽然该项目将通过使用小鼠模型来研究IL 10的调节，但我们的分子研究将由IBD患者GWAS的数据指导。因此，它也应该阐明IL 10在人类疾病中的调节。

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.