Using Multi-Omics to Define Regulators and Drivers of Granulomatous Inflammation and Chronic Beryllium Disease

使用多组学来定义肉芽肿性炎症和慢性铍病的调节因素和驱动因素

• 批准号: 10569103
• 负责人: LISA A MAIER
• 金额: $ 64.77万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-09 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10569103
• 关键词: ATAC-seq; Alleles; Amino Acids; Antigen Presentation; Antigen-Presenting Cells; Antigens; Berylliosis; Beryllium; Biological Markers; Bronchoalveolar Lavage; CD4 Positive T Lymphocytes; COVID-19; Cell Culture Techniques; Cell Differentiation process; Cell Line; Cells; Cellular Indexing of Transcriptomes and Epitopes by Sequencing; Chromatin; Chronic berylliosis; DNA Methylation; DNA analysis; Data; Development; Disease; Drug Targeting; Environment; Environmental Exposure; Epigenetic Process; Epitopes; Etiology; Exposure to; Flow Cytometry; Future; Gene Expression; Genes; Genetic; Genetic Polymorphism; Genetic Predisposition to Disease; Genomic approach; Glutamic Acid; Goals; Granuloma; Granulomatous; HLA-DPB1 gene; Health; Heritability; Hour; Immune; Immune Response Genes; Immune response; Inflammation; Inhalation; Inherited; Innate Immune Response; Investigation; Life; Lung; Macrophage; Measures; Mediating; Methods; Methylation; Modification; Molecular; Organ; Pathogenesis; Pathogenicity; Pathway interactions; Persons; Population; Positioning Attribute; Predisposition; Prevalence; Proliferating; Proteins; Pulmonary Fibrosis; Regulation; Regulatory Pathway; Risk; Risk Factors; Sampling; Site; Structure of parenchyma of lung; System; T-Lymphocyte; Technology; Testing; Time; Transcriptional Regulation; Validation; Variant; Work; XCL1 gene; airway epithelium; asthmatic airway; biomarker identification; chemokine; cytokine; disorder risk; epigenome; fibrotic lung; granulomatous lung disease; histone modification; member; multiple omics; neoantigens; novel; peripheral blood; recruit; response; single cell sequencing; targeted treatment; transcriptome; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT: The goal of this study is to define cell-specific regulatory networks and key drivers of cellular response to beryllium (Be) that result in the granulomatous lung disease, chronic beryllium disease (CBD), at the site of organ involvement. Relying on the complementary, unique expertise of investigative team we will define pathogenic pathways and risk factors for CBD, its precursor (Be sensitization; BeS) and similar environmentally induced diseases. Exposure to an inhaled Be antigen(s), in the setting of a genetically susceptible host, initiates a Th1 immune response, with antigen presentation occurring via HLA Class II on antigen presenting cell (APC) in the context of CD4+ T cells. Subsequently, CD4+ T cells and APCs are recruited to the lung, proliferate, produce cytokines and chemokines, and eventually form granulomas. An increased prevalence of HLA-DPB1 alleles with a glutamic acid at amino acid position 69 (E69) is found in CBD and BeS, although this variant is found in up to 40% of Be exposed workers without BeS or CBD, suggesting that other factors or forms of genetic regulation are important in disease pathogenesis. Growing data in other immune-mediated diseases suggests that epigenetic mechanisms in combination with genetic susceptibility and environment may help explain disease risk. Epigenetic modifications determine T cell and APC/macrophage differentiation and the ensuing immune response through DNA methylation and histone modifications of key genes and thus impact health and disease. Our previous work demonstrated DNA methylation changes associated with alterations in gene expression and disease state in bronchoalveolar lavage (BAL) cells of CBD subjects compared to BeS and controls, including pivotal immune response genes and networks. Single cell sequencing technologies have emerged as a key method to characterize novel cell population and cell-specific changes in gene expression. Based on this information, our hypothesis is that exposure to Be alters epigenetic marks, impacting gene expression and immune cell differentiation in cell-specific manner, and ultimately risk of granulomatous lung disease. We will test this hypothesis through three specific aims, focusing mainly on macrophages and CD4+ T cells but acknowledging that other cell populations are important and considering them in alternative approaches and future directions. In Aim 1 we will characterize the lung immune cell response to Be exposure using single cell ATAC-seq and CITE- seq to measure chromatin accessibility, gene expression levels and protein epitopes at four timepoints to define key drivers in specific cell populations. In Aim 2 we will validate the regulatory networks in uncultured lung samples, focusing on macrophages and CD4+ Tcells, using bulk ATAC- Me and RNA-sequencing. We will functionally validate the results from our multi-omic analysis from Aims 1 and 2 using CRSIPR-dCas9 to change methylation, chromatin accessibility, and gene expression at loci identified as key drivers of response to Be in cultured macrophage and T cell lines in Aim 3. Ultimately, this proposal will enhance our understanding of the novel genes, regulatory pathways and networks, and molecular mechanisms involved in CBD etiology.

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