KLF10 regulates colitis through mediating TGFb induction of FOXP3 in Treg cells.

KLF10 通过介导 Treg 细胞中 TGFb 诱导 FOXP3 来调节结肠炎。

• 批准号: 8100826
• 负责人: William A Faubion
• 金额: $ 19.71万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-15 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8100826
• 关键词: Address; Adoptive Transfer; Affect; Antigens; Area; Autoimmune Diseases; Binding; Biology; Cancer Prognosis; Cell Line; Cells; Chromatin; Chronic; Clinic; Colitis; Colorectal Cancer; Crohn' s disease; DNA; Data; Development; Diabetes Mellitus; Disease; Environment; Enzymes; Epigenetic Process; Equilibrium; Event; Experimental Designs; Experimental Models; Family; Family member; Gene Activation; Gene Expression; Gene Expression Regulation; Gene Targeting; Generations; Genes; Genetic Transcription; Genome; Genomics; Goals; Histone Acetylation; Histones; Human; Immune; Immune response; Immunology; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Intestines; Laboratories; Luciferases; Malignant Neoplasms; Mediating; Methodology; Mus; Neoplasms; Pathogenesis; Pathway interactions; Predisposition; Prevalence; Process; Protein Family; Proteins; Public Health; Qualifying; Recruitment Activity; Regulation; Regulatory Pathway; Regulatory T-Lymphocyte; Reporter; Research Personnel; Rheumatoid Arthritis; Role; Severities; System; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Therapeutic; Transfection; Transgenic Animals; Transgenic Organisms; Work; Zinc Fingers; base; body system; cell type; chromatin modification; chromatin remodeling; cytokine; design; experience; lifetime risk; member; novel; prevent; programs; promoter; resistance mechanism; transcription factor

DESCRIPTION (provided by applicant): The protein FOX3 acts on the DNA of T cells important to immune regulation. Researchers have suspected that this critical FOXP3+ T cell is important in at least the top three most common human autoimmune diseases and the four most deadly cancers. As there are over 8.5 million U.S. residents with an autoimmune disease, it is easy to appreciate the importance of understanding the mechanisms behind the development of this critical FOXP3+ T cell. Our long-term goal is to understand how the cytokine TGF, an extremely important cytokine to the intestinal environment, regulates the generation of these FOXP3+ T cells in the intestine. Our laboratory has discovered that the deficiency of a KLF family member in mice results in a block in intestinal FOXP3+ T cell generation and a tendency to develop colitis similar to human inflammatory bowel disease. Thus, our objective in this application is to determine the mechanism by which KLF family proteins mediate TGF-dependent Foxp3 gene transcription, and how disruption of this pathway leads to colitis. The three specific aims designed to achieve this objective include: (1) Test the hypothesis that KLF10 expression in T cells mediates a mechanism of resistance to colitis, (2) Test the hypothesis that KLF10 regulates distinct genomic control regions of Foxp3, and (3) Test the hypothesis that KLF10 works via distinct chromatin remodeling required for Foxp3 activation. We will use the KLF10-deficient mouse colony and adoptive transfer of immune cell compartments to determine the precise cell type in which KLF10 is critically required to prevent colitis. Subsequent TCR transgenic animals will be used to determine the mechanism by which KLF10 regulates the development of intestinal FOXP3+ T regulatory cells specific for a defined gut antigen. Human T cells lines and transient transfection methodology will be used to characterize the specific regulatory domains of KLF10 required for Foxp3 promoter function. Finally, a novel human T cell line with a defined Foxp3 promoter and luciferase reporter construct stably integrated into the host cell genome will be used to characterize KLF10-dependent chromatin modifications required for Foxp3 promoter activation. This project is significant because upon conclusion, we will understand the role for KLF10 in the regulation of Foxp3, elucidating pathobiological information into the pathogenesis of chronic inflammation and neoplasia. This in turn will stimulate new areas for experimental therapeutics in human chronic inflammatory diseases. PUBLIC HEALTH RELEVANCE: The public health burden of autoimmune disease is significant as these unregulated immune responses affect widely disparate organ systems causing such diseases as inflammatory bowel disease, rheumatoid arthritis and diabetes mellitus. This proposal addresses the mechanisms of development of T regulatory cells, cells expected to protect against autoimmune disease. Results from this proposal will stimulate new areas for experimental therapeutics in human chronic inflammatory diseases.

描述(由申请人提供)：Fox3蛋白作用于T细胞的DNA，对免疫调节至关重要。研究人员怀疑，这种关键的FOXP3+T细胞至少在人类最常见的三种自身免疫性疾病和四种最致命的癌症中是重要的。由于有超过850万美国居民患有自身免疫性疾病，因此很容易理解了解这种关键的FOXP3+T细胞发展背后的机制的重要性。我们的长期目标是了解对肠道环境极其重要的细胞因子转化生长因子如何调节肠道中FOXP3+T细胞的生成。我们的实验室发现，小鼠体内KLF家族成员的缺失会导致肠道FOXP3+T细胞生成受阻，并有发生类似人类炎症性肠病的结肠炎的倾向。因此，我们在这项应用中的目标是确定KLF家族蛋白介导转化生长因子依赖的Foxp3基因转录的机制，以及这一途径的中断如何导致结肠炎。为实现这一目标而设计的三个具体目标包括：(1)测试KLF10在T细胞中的表达介导结肠炎抵抗机制的假设，(2)测试KLF10调节Foxp3不同基因组控制区的假设，以及(3)测试KLF10通过Foxp3激活所需的不同染色质重塑发挥作用的假设。我们将使用KLF10缺乏的小鼠克隆和免疫细胞隔间的过继转移来确定KLF10预防结肠炎所必需的确切细胞类型。随后的TCR转基因动物将被用来确定KLF10调节特定肠道抗原的肠道FOXP3+T调节细胞发育的机制。我们将使用人类T细胞系和瞬时转染法来确定Foxp3启动子功能所需的KLF10的特定调控结构域。最后，一个定义了Foxp3启动子和荧光素酶报告结构稳定整合到宿主细胞基因组中的新型人类T细胞株将被用来表征激活Foxp3启动子所需的依赖于KLF10的染色质修饰。这个项目意义重大，因为在结论的基础上，我们将了解KLF10在Foxp3调控中的作用，阐明慢性炎症和肿瘤发病机制中的病理生物学信息。这反过来将刺激人类慢性炎症性疾病的实验疗法的新领域。 公共卫生相关性：自身免疫性疾病的公共健康负担很大，因为这些不受监管的免疫反应影响到广泛不同的器官系统，导致炎症性肠病、类风湿性关节炎和糖尿病等疾病。这项建议解决了T调节细胞的发展机制，T调节细胞有望预防自身免疫性疾病。这项提议的结果将刺激人类慢性炎症性疾病的实验疗法的新领域。