C-Type Lectins and Immune Surveillance in ALD

C 型凝集素与 ALD 中的免疫监视

• 批准号: 10925432
• 负责人: Adam Kim
• 金额: $ 16.6万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10925432
• 关键词: ATAC-seq; Acute; Address; Affect; Agonist; Alcohol consumption; Alcoholic Hepatitis; Alcoholic Liver Diseases; Alcohols; Bacteria; Bioinformatics; Biological; Biological Assay; Blood; Blood Circulation; C Type Lectin Receptors; C-Type Lectins; CD14 gene; Cell membrane; Cells; Chromatin; Chromatin Structure; Circulation; Clinical; Clinical Immunology; Complement; Data; Dedications; Dendritic Cells; Development Plans; Disease; Disease Progression; Elements; Ethanol Metabolism; Extravasation; FCGR3B gene; Faculty; Family; Focus Groups; Functional disorder; Fungal Components; Future; Gene Cluster; Gene Expression; Gene Expression Regulation; Gene Family; Genes; Genetic Transcription; Genome; Gram-Negative Bacteria; Hypersensitivity; Immune; Immune response; Immune system; Immunologic Surveillance; Immunology; Inflammasome; Inflammation; Inflammatory; Innate Immune Response; Innate Immune System; Leadership; Learning; Lectin Receptors; Ligands; Lipopolysaccharides; Liver; Measurement; Measures; Mediating; Mentors; Microbe; Molecular; Monitor; Morbidity - disease rate; Myelogenous; Pathway interactions; Patients; Pattern; Pattern recognition receptor; Peripheral; Peripheral Blood Mononuclear Cell; Persons; Positioning Attribute; Postdoctoral Fellow; Receptor Gene; Receptor Signaling; Regulation; Research; Ribosomal RNA; Role; Severities; Severity of illness; Signal Transduction; Stimulus; TLR4 gene; Technology; Testing; Toll-like receptors; Transcriptional Regulation; Transposase; Up-Regulation; Variant; Virus; Work; bioinformatics tool; career development; cell type; chronic alcohol ingestion; cytokine; data analysis pipeline; dectin 1; disease phenotype; experimental study; fungus; gut microbes; gut microbiome; gut microbiota; immunological diversity; immunoregulation; inflammatory milieu; inflammatory modulation; liver inflammation; liver injury; machine learning algorithm; member; microbial; microbial products; microorganism interaction; monocyte; mortality; mouse dectin-2; mouse model; mycobiome; new therapeutic target; particle; pathogen; pathogenic bacteria; pathogenic fungus; prevent; receptor expression; response; single-cell RNA sequencing; skills; therapeutic target; tool; transcription factor

PROJECT SUMMARY The Candidate is a postdoctoral fellow committed to developing an academic research group focused on applying bioinformatics to understand the pathophysiology of alcohol-related liver diseases (ALD). His previous and current postdoctoral work has given him the unique skillset to answer questions involving gene expression/regulation in the immune system. The Career Development Plan describes 2 years of mentored research wherein the candidate will develop skills in clinical immunology, generate the sequencing data outlined in the proposal, and learn leadership skills to transition into independence. The next 3 years, after obtaining an independent faculty position, will be dedicated to developing new data analysis pipelines and establishing cell biological and bioinformatics tools to understand gene regulation in the innate immune system, which will allow the candidate to establish future projects in ALD immunology. Research Plan: ALD is a spectrum of disorders that affect a growing number of people worldwide. Alcoholic Hepatitis (AH) is a severe inflammatory disease that can increase the morbidity and severity of ALD disorders. In AH, the innate immune system is hypersensitive to microbial byproducts. Alcohol consumption causes gut-barrier disruption, leading to leakage of gut microbes into the portal circulation. The liver immune system is able to detect these microbes through pattern recognition receptors, including the Toll-like receptors (TLRs) and C-type Lectin receptors (CTLs). While TLRs have been well studied for their role in sensitizing the innate immune cells to microbial products, the CTLs have only recently been implicated in mouse models of ALD. CTLs are a large family of PRRs that sense a vast diversity of microbes, including bacteria, fungi, and viruses. Our work has found that many members of the CTL gene family are upregulated in the liver and peripheral blood mononuclear cells PBMCs of AH patients. CTLs were also upregulated in PBMCs in response to LPS/TLR4 signaling. We predict CTLs are upregulated in response to gut- derived LPS in order to increase sensing for other microbes that may be present in the blood, making this pathway a secondary innate immune surveillance pathway. In this proposal, we will test this hypothesis in three aims to address the functional role and regulation of this immune surveillance pathway, and the mechanism by which it is exacerbated in AH. First, we will use PBMCs isolated from AH patients to measure increased sensitivity to CTL agonists, as well as micro-/mycobiome sequencing to determine what microbial byproducts were in circulation in patients. Second, we will use single-cell RNA sequencing (scRNA-seq) to dissect the different monocyte subclasses, variation in CTL expression, and how different cell types respond to CTL agonists. Third, because CTL genes are clustered in the genome, we will use scRNA-seq and ATAC-seq (Assay for Transposase-Accessible Chromatin) to understand co-regulation of nearby genes in the genome. Altogether, this proposal will further our understanding of CTL mediated immune surveillance in host/microbial interactions during AH disease progression and potentially identify new therapeutic targets to decrease inflammation.

项目摘要 候选人是致力于建立一个专注于的学术研究小组的博士后研究员 应用生物信息学以了解酒精相关肝病（ALD）的病理生理。他的前任 当前的博士后工作为他提供了独特的技能，可以回答涉及基因的问题 免疫系统中的表达/调节。职业发展计划描述了2年的指导 研究候选人将发展临床免疫学的技能，生成概述的测序数据 在提案中，学习领导能力以过渡到独立性。接下来的3年，获得了 独立的教师职位将致力于开发新的数据分析管道并建立细胞 了解先天免疫系统中基因调节的生物学和生物信息学工具，这将允许 在ALD免疫学领域建立未来项目的候选人。研究计划：ALD是一系列疾病 这会影响全球越来越多的人。酒精性肝炎（AH）是一种严重的炎症性疾病 可以增加ALD疾病的发病率和严重程度。在AH中，先天免疫系统对 微生物副产品。饮酒会导致肠道障碍，导致肠道微生物泄漏到 门户流通。肝免疫系统能够通过模式识别检测这些微生物 受体，包括Toll样受体（TLR）和C型凝集素受体（CTL）。而TLR已经 对它们在使先天性免疫细胞对微生物产物敏感的作用方面进行了良好研究，CTL直到最近才有 与ALD的小鼠模型有关。 CTL是一个大型的PRR家族，感知到大量的多样性 微生物，包括细菌，真菌和病毒。我们的工作发现CTL基因家族的许多成员 在AH患者的肝脏和外周血单核细胞PBMC中被上调。 CTL也是 响应LPS/TLR4信号传导，在PBMC中上调。我们预计CTL会响应肠道上调 派生的LP，以增加对血液中可能存在的其他微生物的感应，从而使这 途径是二级先天免疫监视途径。在此提案中，我们将在三个中检验这一假设 旨在解决该免疫监视途径的功能作用和调节，以及通过 它在ah中加剧了。首先，我们将使用与AH患者分离的PBMC测量增加 对CTL激动剂的敏感性以及微生物组测序，以确定哪种微生物副产品 患者流通。其次，我们将使用单细胞RNA测序（SCRNA-SEQ）进行剖析 不同的单核细胞亚类，CTL表达的变化以及不同的细胞类型对CTL的反应 激动剂。第三，由于CTL基因聚集在基因组中，因此我们将使用SCRNA-SEQ和ATAC-SEQ（测定法 用于转座酶可访问的染色质）了解基因组中附近基因的共同调节。共， 该建议将进一步了解宿主/微生物相互作用中CTL介导的免疫监视 在AH疾病进展过程中，并有可能识别新的治疗靶标，以减少炎症。