C-Type Lectins and Immune Surveillance in ALD

C 型凝集素与 ALD 中的免疫监视

• 批准号: 10683563
• 负责人: Adam Kim
• 金额: $ 24.88万
• 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 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10683563
• 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; Clinical; Clinical Immunology; Complement; Data; 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 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; Lead; Leadership; Learning; 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; base; 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; host-microbe interactions; immunological diversity; immunoregulation; inflammatory milieu; liver inflammation; liver injury; machine learning algorithm; member; microbial; monocyte; mortality; mouse dectin-2; mouse model; mycobiome; new therapeutic target; particle; pathogen; pathogenic bacteria; pathogenic fungus; pathogenic virus; 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.

项目概要 候选人是一名博士后研究员，致力于发展一个专注于以下领域的学术研究小组 应用生物信息学来了解酒精相关性肝病（ALD）的病理生理学。他之前的 目前的博士后工作使他拥有独特的技能来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，其中候选人将培养临床免疫学技能，生成概述的测序数据 在提案中，学习领导技能以过渡到独立。获得资格后的未来3年 独立的教职职位，将致力于开发新的数据分析管道和建立细胞 生物学和生物信息学工具来了解先天免疫系统中的基因调控，这将允许 建立 ALD 免疫学未来项目的候选人。研究计划：ALD 是一系列疾病 影响着全世界越来越多的人。酒精性肝炎（AH）是一种严重的炎症性疾病， 会增加 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。饮酒会导致肠道屏障破坏，导致肠道微生物渗漏到体内。 门户循环。肝脏免疫系统能够通过模式识别来检测这些微生物 受体，包括 Toll 样受体 (TLR) 和 C 型凝集素受体 (CTL)。虽然 TLR 已 CTL 在使先天免疫细胞对微生物产物敏感方面的作用得到了充分研究，直到最近才被 与 ALD 小鼠模型有关。 CTL 是 PRR 的一个大家族，可以感知大量的多样性 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 AH 患者的肝脏和外周血单核细胞 PBMC 中表达上调。 CTL 还 PBMC 响应 LPS/TLR4 信号而上调。我们预测 CTL 会因肠道反应而上调 衍生的 LPS，以增强对血液中可能存在的其他微生物的感应，从而使 途径 次级先天免疫监视途径。在这个提案中，我们将通过三个方面来检验这个假设 旨在解决这种免疫监视途径的功能作用和调节及其机制 这种情况在 AH 中更加严重。首先，我们将使用从 AH 患者分离的 PBMC 来测量增加的 对 CTL 激动剂的敏感性，以及微生物/真菌组测序以确定哪些微生物副产物 已在患者体内循环。其次，我们将使用单细胞 RNA 测序 (scRNA-seq) 来剖析 不同的单核细胞亚类、CTL 表达的变化以及不同细胞类型对 CTL 的反应 激动剂。第三，由于CTL基因在基因组中聚集，我们将使用scRNA-seq和ATAC-seq（Assay 用于转座酶可及的染色质）来了解基因组中附近基因的共同调节。共， 该提案将进一步加深我们对宿主/微生物相互作用中 CTL 介导的免疫监视的理解 在 AH 疾病进展期间，并可能确定新的治疗靶点以减少炎症。