Genetic regulation of intestinal epithelial cell innate immune signaling in human type 1 diabetes

人类1型糖尿病肠上皮细胞先天免疫信号的基因调控

• 批准号: 10047614
• 负责人: Shannon Margaret Wallet
• 金额: $ 7.23万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-05 至 2020-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10047614
• 关键词: Genetic; regulation; intestinal; epithelial; cell

Project Abstract/Summary Type 1 Diabetes (T1D) is an autoimmune disease with both genetic and environmental contributions, yet a single environmental factor has yet to be identified. We propose that rather than the specific identity of the environmental factor, the way in which an environmental signal is received is most important to disease. Among individuals at risk of developing T1D, seroconversion to islet autoantibody (IA) production is the best and earliest biological predictor of disease progression. Thus, critical events leading to the destructive β cell autoimmunity associated with T1D are linked to the development of IAs. In subjects with high risk HLA alleles, 9 gene regions marked by single nucleotide polymorphisms (SNPs) are associated with IA seroconversion. Our collaborator’s analysis of these IA risk loci reveals enrichment for genes/proteins that regulate only three cell signaling pathways (RAS/MAPK, PI3K/AKT and JAK/STAT), whereby four IA seroconversion risk variants (PTPN22, SH2B3, ERBB3 and UBASH3A) each impact all three. Thus, we propose that regulation of environmental sensing is at least in part controlled by the one or more of these risk variants associated with the development of IAs. Appropriate communication of the gastrointestinal tract (GI) with the environment is required for local and systemic immunologic homeostasis, which is in part regulated by the intestinal epithelial cell (IEC) through their physical and biochemical interactions with innate and adaptive immune populations. To this end, contribution of immunity within the GI tract to the expression of autoimmunity at distal sites has been previously demonstrated. Thus we propose that the seroconversion risk variants affect immune signaling pathways within the IEC contributing to the maintenance of autoimmunity in T1D, through the secondary expansion and/or polarization of autoreactive effector T-cells. To study human disease, a major need is cell-specific human isogenic systems that can be used to dissect the role of individual T1D risk gene variant alleles. Our preliminary studies support use of a disease-in- a-dish strategy that pairs human induced pluripotent stem cells (iPSCs) with CRISPR/Cas9 gene modification to fulfill these needs. We propose to couple iPSCs and CRISPR/Cas9 technology with our method of reproducible primary human IECs culture to achieve haploinsufficient, mono-allelic homozygosity at risk or protective alleles to evaluate alterations in IEC-immune mechanisms which contribute to the maintenance of βcell autoimmunity.

项目摘要/摘要 1型糖尿病（T1D）是一种具有遗传和环境贡献的自身免疫性疾病 单个环境因素尚未确定。我们建议，而不是特定的身份 环境因素，接收环境信号的方式对疾病最重要。之中 有发育T1D，胰岛自动抗体（IA）生产的人的风险是最好，最早的 疾病进展的生物学预测指标。这是导致破坏性β细胞自身免疫性的关键事件 与T1D相关的与IAS的发展有关。在高风险HLA等位基因的受试者中，9个基因区域 以单核苷酸多态性（SNP）为特征与IA血清转化有关。我们的合作者 对这些IA风险当地人的分析揭示了仅调节三个细胞信号的基因/蛋白质的富集 途径（RAS/MAPK，PI3K/AKT和JAK/STAT），其中四个IA Seroconversion风险变体（PTPN22，PTPN22， SH2B3，ERBB3和UBASH3A）每个都撞击这三个。那，我们提出了对环境的调节 感知至少部分由与开发相关的这些风险变体中的一个或多个控制 IAS。 在本地和 系统性免疫稳态，部分由肠上皮细胞（IEC）调节 与先天和适应性免疫吞噬的物理和生化相互作用。为此， 先前已经证明了胃肠道对自身免疫性表达的自身免疫表达的免疫力。 我们建议血清转化风险变体影响IEC内的免疫信号通路 通过次要扩展和/或极化，有助于维持T1D自身免疫性 自动反应效应子T细胞。 为了研究人类疾病，主要需求是细胞特异性的人源系统，可用于 剖析单个T1D风险基因变异等位基因的作用。我们的初步研究支持使用疾病 A-dish策略将人类诱导多能干细胞（IPSC）与CRISPR/CAS9基因修饰诱导的策略 满足这些需求。我们建议将IPSC和CRISPR/CAS9技术与我们的可再现方法相结合 初级人类IEC培养 评估IEC免疫机制的改变，这有助于维持βcell自身免疫性。