Chromosome 17q21, allergic inflammation, and remodeling

染色体 17q21、过敏性炎症和重塑

基本信息

批准号：
10581533
负责人：
DAVID H BROIDE
金额：
$ 56.14万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-05-15 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10581533
关键词：
17q21 Alleles Allergens Allergic inflammation Applications Grants Asthma Biological Biology Blood Cells CD4 Positive T Lymphocytes Caring Cell Adhesion Cell Proliferation Cell physiology Cells Chemotaxis Childhood Chromosomes Development Funding Genes House Dust House Dust Mite Allergens Human Hypersensitivity Immune Immunology In Vitro Individual Laboratories Link Lung Mediating Mus Nature Pathogenesis Pathway interactions Play Population Heterogeneity Production Proteomics Pyroglyphidae Role Science Th2 Cells Transgenic Mice Vascular Endothelial Cell airway remodeling asthma model asthmatic base editing cytokine ethnic diversity genetic association genome wide association study in vivo insight knock-down mouse model novel overexpression polarized cell postnatal postnatal period respiratory smooth muscle response risk variant selective expression trafficking transcriptome sequencing

项目摘要

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.

该提案的重点是染色体 17q21 上的 ORMDL3 基因，该基因与多项全基因组关联研究中的人类哮喘。由于 ORMDL3 在 CD4+ 中高表达细胞，在本提案中，我们试图确定 ORMDL3 如何影响 CD4+ 细胞功能、过敏炎症、哮喘。尽管 17q21 号染色体与哮喘的关联已得到充分证实，支持与发病机制相关的肺细胞中这种关联的生物学机制哮喘尚不为人们所了解，这也是本提案的重点。另外因为至少有9个基因位于染色体 17q21 上的这个区域，了解每个基因的生物学特性对于了解该区域如何影响哮喘的发展非常重要。作为 SNP 连接 ORMDL3 与哮喘的关系与 ORMDL3 表达水平的增加有关，我们生成了通用的 ORMDL3 转基因 (TG) 小鼠表达人类 ORMDL3 (hORMDL3) 和证明它们自发地（在没有接触过敏原的情况下）显着发育 ASM 增加和 AHR 增加是哮喘的主要特征。除了这一重要的基线效应之外 ORMDL3 在 ASM 中对 AHR 的影响，ORMDL3 在增强 Th2 反应和 AHR 方面也发挥着重要作用正如我们对过敏原攻击的通用 hORMDL3 TG 小鼠的研究所证明的那样。因此，重点该提案是为了增加我们对 ORMDL3 在 CD4+ T 淋巴细胞中如何表达的理解增强 Th2 对过敏原挑战的反应。在本拨款提案中，我们建议证明表达 ORMDL3 水平升高的 CD4+ 细胞（小鼠和人类）可增强 Th2 反应体外和体内室内尘螨 (HDM) 过敏原，并使用蛋白质组学和 RNAseq 方法计划以确定 HDM 四聚体阳性 CD4+ 细胞中介导这种 ORMDL3 效应的下游途径。 ORMDL3 下游途径的靶向敲除或过度表达将证明它们对 CD4 细胞功能的贡献。最后，我们使用单碱基编辑来编辑与 ORMDL3 关联的 SNP，以确定哪些 SNP 功能性调节 ORMDL3、Th2 的水平细胞因子和 ORMDL3 的下游途径。