C-Type Lectins and Immune Surveillance in ALD

C 型凝集素与 ALD 中的免疫监视

• 批准号: 10020883
• 负责人: Adam Kim
• 金额: $ 12.31万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-20 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10020883
• 关键词: ATAC-seq; Acute; Address; Affect; Agonist; Alcohol consumption; Alcoholic Hepatitis; 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; Liver diseases; Measurement; Measures; Mediating; Mentors; Microbe; Molecular; Monitor; Morbidity - disease rate; Myelogenous; Pathway interactions; Patients; Pattern; Pattern recognition receptor; Peripheral; Peripheral Blood Mononuclear Cell; 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; 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 候选人是一名博士后研究员，致力于发展一个学术研究小组，重点是 应用生物信息学了解酒精相关性肝病（ALD）的病理生理学。他以前 目前的博士后工作给了他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，其中候选人将发展临床免疫学技能，生成概述的测序数据 在提案中，并学习领导技能，过渡到独立。在接下来的三年里， 独立的教师职位，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，以了解先天免疫系统中的基因调控，这将使 候选人在ALD免疫学建立未来的项目。研究计划：ALD是一系列疾病 影响着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， 可增加ALD病症的发病率和严重性。在AH中，先天免疫系统对 微生物副产物饮酒会破坏肠道屏障，导致肠道微生物渗漏到 门静脉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）。尽管TLR已经被 CTL在使先天免疫细胞对微生物产物敏感方面的作用已得到充分研究，但直到最近才 与ALD的小鼠模型有关。CTL是一个大家族的PRR，其感测多种多样的细胞因子。 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中上调。CTL也 在PBMC中响应LPS/TLR 4信号传导而上调。我们预测CTL在肠道反应中上调- 衍生的LPS，以增加对可能存在于血液中的其他微生物的感知， 途径是二级先天免疫监视途径。在本建议中，我们将在三个方面测试这一假设。 旨在解决这一免疫监视途径的功能作用和调节，以及通过 而这一点，在《易经》中，是不言而喻的。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/真菌生物群测序，以确定哪些微生物副产物 在病人体内循环第二，我们将使用单细胞RNA测序（scRNA-seq）来分析 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基因在基因组中成簇，我们将使用scRNA-seq和ATAC-seq（测定 转座酶可降解染色质），以了解基因组中邻近基因的共同调控。总的来说， 这一建议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疾病进展过程中，并可能确定新的治疗靶点以减少炎症。