Project 2

项目2

基本信息

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
项目类别：
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

项目摘要

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 探索了范式转换假设，即上皮细胞是两种细胞的核心。严重哮喘（SA）的分子表型根本不同。第一，支气管上皮细胞（BEC）对初始环境刺激做出反应，驱动先天的内在表型。第二，BEC 细胞死亡途径与 CD8 T 细胞免疫过程交叉，驱动免疫相互作用表型。该假设基于已发表、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，一种独立于 T 细胞，另一种与 T 细胞高度相关，尤其是与 CD8 细胞。两者中都存在相似水平的 2 型 (T2) 生物标志物/基因。在已发表的上一个周期的数据显示，上皮生长和修复模块与哮喘的严重程度高于 T2 模块，表明 T2 免疫本身并不会导致严重程度。据报道，与基因表达一致，哮喘 BEC 的体外伤口修复不良，并伴有增殖能力的丧失。附着的粘蛋白 MUC4b 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，每种表型都具有相似水平的 T2 生物标志物。这先天内在/非 T 细胞 SA 簇与先天 NF-kB 和凋亡途径相关可能通过 Toll 样受体 (TLR)。免疫相互作用/CD8簇关联与 IL-12 和 -18、干扰素以及炎症小体和焦亡途径的上调通过 17q12-21 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体外和体内模型，我们将完善分子在第一个周期中确定的表型，并将它们与项目 1 中确定的细胞概况联系起来。我们提出 3 个目标：1）确定先天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/CD8 相关免疫相互作用表型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，作为倒数第二项努力连接细胞类型、机制和区室以严格细化表型。这些研究将提高对与细胞死亡相关的分子表型的理解，从而使识别新的精准治疗靶点，最终确认潜在的内型。