Chromosome 17q21, allergic inflammation, and remodeling

这项提议的焦点是位于染色体17q21上的ORMDL3基因，该基因与 几项全基因组关联研究中的人类哮喘。由于ORMDL3在CD4+细胞中高表达 细胞，在这项建议中，我们试图确定ORMDL3如何影响CD4+细胞的功能，过敏 炎症和哮喘。尽管染色体17q21与哮喘的联系已经很好地建立起来了， 肺细胞中这种联系的生物学机制(S)与慢性阻塞性肺疾病的发病机制有关 哮喘没有得到很好的理解，也是这项提案的重点。此外，由于至少有9个基因 位于染色体17q21上的这一区域，分别了解这些基因的生物学特性 对于了解该区域如何影响哮喘的发展很重要。作为SNP链接 ORMDL3与哮喘相关，我们产生了普遍的ORMDL3表达水平增加 ORMDL3转基因(TG)小鼠，表达增加的人ORMDL3(HORMDL3)和 证明它们自发地(在没有过敏原暴露的情况下)显著发展 哮喘的主要特征是ASM增加和AHR增加。除了这一重要的基线效应之外 ORMDL3在ASM中对AHR的作用也在增强Th2应答和AHR方面发挥重要作用 正如我们对过敏原挑战通用hORMDL3TG小鼠的研究所表明的那样。因此，重点是 这项建议是为了增加我们对ORMDL3如何在CD4+T淋巴细胞中表达的理解 增强Th2细胞对变应原挑战的反应。在这份赠款提案中，我们打算证明 表达高水平ORMDL3的CD4+细胞(小鼠和人)增强了Th2反应 体外和体内对尘螨(HDM)变应原的研究，并使用蛋白质组学和RNAseq方法计划 目的：确定HDM四聚体阳性的CD4+细胞中介导ORMDL3效应的下游通路。 靶向敲除或过度表达被确定为ORMDL3下游的通路将 展示它们对CD4细胞功能的贡献。最后，我们正在使用单碱基编辑来编辑 与ORMDL3连锁的SNP确定哪些SNP在功能上调节ORMDL3、Th2的水平 细胞因子和ORMDL3的下游途径。

The focus of this proposal is on ORMDL3 a gene on chromosome 17q21, which has been highly linked to human asthma in several genome wide association studies. As ORMDL3 is highly expressed in CD4+ cells, in this proposal we seek to determine how ORMDL3 influences CD4+ cell function, allergic inflammation, and asthma. Although the linkage of chromosome 17q21 to asthma is very well established, the biologic mechanism(s) underpinning this association in lung cells pertinent to the pathogenesis of asthma is not as well understood and is the focus of this proposal. In addition as there are at least 9 genes located in this region on chromosome 17q21 understanding the biology of each of these genes individually is important to understanding how this region influences the development of asthma. As the SNP linking ORMDL3 to asthma is associated with increased levels of ORMDL3 expression, we generated universal ORMDL3 transgenic (TG) mice that express increased levels of human ORMDL3 (hORMDL3) and demonstrated that they spontaneously (in the absence of allergen exposure) develop significantly increased ASM and increased AHR, major features of asthma. In addition to this important baseline effect of ORMDL3 in ASM on AHR, ORMDL3 also plays a significant role in enhancing Th2 responses and AHR as demonstrated in our studies of allergen challenged universal hORMDL3 TG mice. Thus, the focus of this proposal is to increase our understanding of how ORMDL3 expressed in CD4+ T lymphocytes enhances Th2 responses to allergen challenge. In this grant proposal we propose to demonstrate that CD4+ cells (mouse and human) expressing increased levels of ORMDL3 have enhanced Th2 responses in vitro and in vivo to house dust mite (HDM) allergen, and using proteomic and RNAseq approaches plan to identify downstream pathways in HDM tetramer positive CD4+ cells that mediate this ORMDL3 effect. Targeted knockdown or overexpression of pathways identified to be downstream of ORMDL3 will demonstrate their contribution to the function of CD4 cells. Finally, we are using single base editing to edit SNPs linked to ORMDL3 to determine which SNPs functionally regulate levels of ORMDL3, Th2 cytokines, and downstream pathways of ORMDL3.