Project 2

项目2

• 批准号: 10625519
• 负责人: Sally E Wenzel
• 金额: $ 53.67万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10625519
• 关键词: 17q12; 17q21; Adverse event; Animal Model; Apoptotic; Asthma; Bioinformatics; Biological Markers; CCL26 gene; CD8-Positive T-Lymphocytes; CD8B1 gene; Cell Death; Cell Survival; Cell physiology; Cells; Cessation of life; Clinical; Coculture Techniques; Data; Development; ERBB2 gene; Elements; Epigenetic Process; Epithelial Cell Proliferation; Epithelial Cells; Epithelium; Functional disorder; Gene Cluster; Gene Expression; Genes; Genetic; Growth; Growth Factor Receptors; Heart; Heterogeneity; IL18 gene; Immune; Immune response; Immunity; In Vitro; Inflammasome; Inflammatory; Interferons; Interleukin-12; Ligands; Link; Lymphocyte; MUC5AC gene; Methylation; Mitosis; Molecular; Mucin 1 protein; Mucins; NF-kappa B; Natural Immunity; Neurons; Outcome; Pathway interactions; Patients; Phenotype; Precision therapeutics; Process; Proliferating; Protein Analysis; Proteins; Publishing; Refractory; Reporting; Research; Risk; Role; Severities; Steroids; Stimulus; System; T-Lymphocyte; Techniques; Tissues; Toll-Like Receptor Pathway; Toll-like receptors; Up-Regulation; Validation; adaptive immunity; airway epithelium; asthmatic; bronchial epithelium; cell type; clinical phenotype; cohort; computerized tools; genome wide association study; genomic locus; immune activation; improved; in vivo; in vivo Model; molecular phenotype; new therapeutic target; novel; periostin; peripheral blood; programs; repaired; response; sialylation; single cell technology; single-cell RNA sequencing; targeted treatment; therapeutic target; transcriptome sequencing; treatment response; wound healing

Project 2 explores the paradigm-shifting hypothesis that the epithelium is at the heart of two fundamentally different molecular phenotypes of severe asthma (SA). In the 1st, bronchial epithelial cells (BEC)s respond to initial environmental stimuli to drive an innate intrinsic phenotype. In the 2nd, BEC cell death pathways intersect with CD8 T-cell immune processes to drive an immune interactive phenotype. This hypothesis is based on published, submitted and preliminary data across Project 1 and Project 2 which identify two immune cell phenotypes of severe asthma, one independent of T-cells and another, highly associated with them, in particular with CD8 cells. Similar levels of Type-2 (T2) biomarkers/genes are present in both. In published data from the previous cycle, epithelial growth and repair modules more strongly associated with asthma severity than T2 modules, suggesting T2 immunity alone does not drive severity. Consistent with gene expression, poor wound repair of asthmatic BECs in vitro was reported, with associated loss of a proliferative capacity. The tethered mucin MUC4b, increased in asthma, lowers BEC proliferation, while increasing secretion. BEC gene clustering mirrors Project 1 with T-cell- and non-T-cell associated SA phenotypes, each with similar levels of T2 biomarkers. The innate intrinsic/Non-T-cell SA cluster associates with innate NF-kB and apoptotic pathways potentially through Toll like receptors (TLR)s. The immune interactive/CD8 cluster associates with IL-12 and -18, interferons and upregulation of inflammasome and pyroptosis pathways through the 17q12-21 gene, GSDMB. Thus, interactions between BEC genetic/epigenetic risks, innate and adaptive immunity and cell death pathways may contribute to these distinct phenotypes, with differing treatment responses. Using cutting edge technologies of scRNAseq, advanced bioinformatics and mechanistic in vitro and in vivo models, we will refine the molecular phenotypes identified in the 1st cycle and link them with cell profiles identified in Project 1. We propose 3 aims: 1) Identify the mechanisms for and functional implications of an innate intrinsic epithelial phenotype using fresh and cultured healthy and asthmatic BECs 2) Evaluate the role of GSDMB in the development of a IL-18/CD8-associated immune interactive phenotype and its functional implications using fresh and cultured healthy and asthmatic BECs 3) Integrate immune-inflammatory phenotypes with epithelial molecular phenotypes, as the penultimate effort to link cell types, mechanisms and compartments to rigorously refine phenotypes. These studies will improve the understanding of molecular phenotypes in relation to cell death, allowing Identification of novel precision therapeutic targets to eventually confirm underlying endotypes.

项目2探讨了范式转移假说，即上皮细胞是两个细胞的核心。 严重哮喘（SA）的根本不同的分子表型。在第一，支气管 上皮细胞（BEC）对初始环境刺激作出反应，以驱动先天的内在 表型第二，BEC细胞死亡途径与CD 8 T细胞免疫过程交叉， 驱动免疫互动表型。这一假设是基于已发表、提交和 项目1和项目2的初步数据，确定了两种免疫细胞表型， 严重哮喘，一个独立于T细胞，另一个与T细胞高度相关，特别是 CD 8细胞在两者中存在相似水平的2型（T2）生物标志物/基因。于公开的 来自上一周期的数据，上皮生长和修复模块与 哮喘的严重程度高于T2模块，这表明T2免疫本身并不驱动严重程度。 与基因表达一致，据报道，哮喘BEC在体外的伤口修复较差， 伴随着增殖能力的丧失。系留粘蛋白MUC 4 b在哮喘中增加， 降低BEC增殖，同时增加分泌。BEC基因聚类反映了项目1， T细胞和非T细胞相关的SA表型，每种表型具有相似的T2生物标志物水平。的 先天性内在/非T细胞SA簇与先天性NF-kB和凋亡途径相关 可能通过Toll样受体（TLR）。免疫相互作用/CD 8簇相关 IL-12和IL-18、干扰素以及炎性小体和细胞凋亡途径的上调 通过17 q12 -21基因，GSDMB。因此，BEC遗传/表观遗传风险之间的相互作用， 先天性和适应性免疫以及细胞死亡途径可能有助于这些不同的 表型，具有不同的治疗反应。利用scRNAseq的尖端技术， 先进的生物信息学和机制，在体外和体内模型，我们将完善分子 在第一个周期中鉴定的表型，并将它们与项目1中鉴定的细胞谱联系起来。我们 提出3个目标：1）确定先天性内在的机制和功能含义， 使用新鲜和培养的健康和哮喘BEC的上皮表型2）评估作用 GSDMB在IL-18/CD 8相关免疫相互作用表型的发展中的作用， 使用新鲜和培养的健康和哮喘BEC的功能意义3）整合 免疫炎症表型与上皮分子表型，作为倒数第二个努力 将细胞类型、机制和区室联系起来，严格细化表型。这些研究 将提高对细胞死亡相关分子表型的理解， 鉴定新的精确治疗靶点，以最终确认潜在的内型。