Uncover the new subsets of epidermal Langerhans cells

发现表皮朗格汉斯细胞的新亚群

• 批准号: 10020173
• 负责人: Indra Adrianto
• 金额: $ 37.02万
• 依托单位: HENRY FORD HEALTH SYSTEM
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-17 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10020173
• 关键词: Adult; Aging; Antigen-Presenting Cells; Autoimmune Diseases; Autoimmune Process; Bioinformatics; Biological Markers; Biology; Birth; Bone Marrow; C-Type Lectins; Cell Differentiation process; Cell Maturation; Cell model; Cells; Cellular Assay; Cellular biology; Chromatin; Data; Dendritic Cells; Development; Disease model; Embryo; Embryonic Development; Epidermis; Fetal Liver; Goals; Growth; Heterogeneity; Human; Hypersensitivity; Immune; Immune Tolerance; Immunity; Immunologist; In Vitro; Individual; Infection; Inflammation; Inflammatory; Intervention; Knockout Mice; Langerhans cell; Lead; Leishmaniasis; Maps; Mediating; Minor; Modeling; Molecular Profiling; Mus; Mutation; Mycoses; Names; Outcome; Peripheral; Phase; Phenotype; Play; Population; Procedures; Regulatory Element; Regulatory T-Lymphocyte; Reporter; Research; Research Personnel; Resolution; Role; Skin; Skin Cancer; Sorting - Cell Movement; T-Lymphocyte; Testing; Time; Transposase; Variant; Vitiligo; Work; Yolk Sac; activating transcription factor 3; adaptive immunity; base; epigenomics; experience; genetic signature; immune function; immunoregulation; in vivo; insight; langerin; macrophage; melanoma; monocyte; mouse model; new technology; progenitor; single cell analysis; single-cell RNA sequencing; skin disorder; transcriptome; transcriptome sequencing; transcriptomics

PROJECT SUMMARY Langerhans cells (LCs) are skin-resident dendritic cells (DCs) expressing the C-type lectin Langerin (CD207) that mediate both adaptive immunity and immune tolerance in skin and are involved in various types of skin diseases. Adult LCs are originated from embryonic yolk-sac-derived macrophages and fetal liver monocytes in the steady state. Interestingly, LCs could also be derived from the bone marrow or peripheral monocytes and repopulate the skin under inflammatory conditions. However, due to the lack of molecular profiles at individual LC level, a significant gap remains in our understanding on how a single CD207+ epidermal LC population can induce both immunity and tolerance. Fortunately, new technologies such as the single-cell RNA-sequencing (scRNA-seq) can evaluate cell-to-cell transcriptomic variation, while the single-cell assay for transposase- accessible chromatin using sequencing (scATAC-seq) can assess the epigenomic heterogeneity at single-cell resolution in an unbiased manner. Recently, we identified two major LC subsets in mice, ATF3+Bal2a1b- (mLC1) and ATF3-Bal2a1b+(mLC2) subsets, and three major LC subsets in human including ATF3+ (hLC1) subset using scRNA-seq. We also found in ATF3 knockout mice that lack of ATF3 enhances LC maturation and promotes LCs-induced Th1 and Th17 cell differentiation suggesting immune suppressive function induced by ATF3+LC1. Hence, these preliminary data support our hypothesis that LCs are heterogeneous consisting of distinct subsets with different immune functions. Our objective is to use single-cell analysis platforms plus the LC fate-mapping and mutation mouse models to further validate this. We will pursue two Specific Aims in the R61 phase: Aim 1) Characterize the gene signatures and regulatory elements of mLC1 and mLC2 by profiling LCs during embryonic, young, and aging development at steady-state and repopulated LCs at inflamed state using scRNA-seq and scATAC-seq; Aim 2) Generate ATF3negEGFP reporter mice to fate-map ATF3+LC1 embryonic development and the dynamic change of ATF3+LC1 and ATF3-LC2 subset at steady state during adult and aging development and at inflammatory state and functionally characterize LC subsets in vitro by sorting ATF3EGFP+ LC1 and ATF3- LC2 cells and rederiving ATF3.loxp mice, which will be crossed with hLangerin-Cre mice to generate LC- specific/time-induced ATF3KO for in vivo functional study. In the R33 phase, we will pursue the following Specific Aim: Aim 3) Functionally characterize ATF3+LC1 subset in vivo using LC-specific ATF3 deletion hLCcre.ATF3KO mice to evaluate the potential immune regulation function of ATF3+LC1 subset in the different disease models, including autoimmune vitiligo, melanoma, and fungi infection models. Our work will uncover the mystery of LC subsets with their specific functions, which will provide new insights into the biology of LCs and lead to the development of LC-based intervention strategies for skin diseases.

项目摘要 Langerhans细胞（LCS）是表达C型凝集素链霉素的皮肤居民树突状细胞（DCS）（CD207） 介导皮肤的适应性免疫和免疫耐受性，并参与各种类型的皮肤 疾病。成年LC源自胚胎蛋黄 - 萨克衍生的巨噬细胞和胎儿肝单核细胞 稳定状态。有趣的是，LC也可以源自骨髓或外周单核细胞，以及 在炎症条件下重新填充皮肤。但是，由于个人缺乏分子曲线 LC水平，我们对单个CD207+表皮LC种群如何如何才能了解的显着差距 诱导免疫力和耐受性。幸运的是，新技术（例如单细胞RNA） （SCRNA-SEQ）可以评估细胞对细胞转录组的变化，而单细胞测定转座酶 - 使用测序（SCATAC-SEQ）可访问的染色质可以评估单细胞的表观基因组异质性 以公正的方式解决。最近，我们确定了小鼠ATF3+BAL2A1B-（MLC1）的两个主要LC子集 和ATF3-BAL2A1B+（MLC2）子集，以及人类中的三个主要LC子集，包括ATF3+（HLC1）子集 scrna-seq。我们还在ATF3敲除小鼠中发现缺乏ATF3可增强LC成熟并促进 LCS诱导的Th1和Th17细胞分化表明ATF3+LC1诱导的免疫抑制功能。 因此，这些初步数据支持我们的假设，即LC是由不同的子集组成的异质的 具有不同的免疫功能。我们的目标是使用单细胞分析平台以及LC命运映射 和突变小鼠模型以进一步验证这一点。我们将在R61阶段实现两个具体目标：AIM 1） 通过在胚胎期间分析LCS，表征MLC1和MLC2的基因特征和调节元件 使用scrna-seq和 scatac-seq;目的2）生成ATF3NegeGFP记者小鼠以命运映射ATF3+LC1胚胎开发和 成人和衰老期间，ATF3+LC1和ATF3-LC2子集的动态变化 在炎症状态下，通过对ATF3EGFP+ LC1和ATF3-进行排序，在功能上表征了LC子集的体外表征 LC2细胞和Rederate Atf3.loxp小鼠，将与Hlangerin-Cre小鼠交叉以产生LC- 特定/时间诱导的ATF3KO用于体内功能研究。在R33阶段，我们将追求以下特定的特定 AIM：AIM 3）使用LC特异性ATF3删除HLCCRE.ATF3KO在功能上表征ATF3+LC1子集 小鼠评估不同疾病模型中ATF3+LC1子集的潜在免疫调节功能， 包括自身免疫性白癜风，黑色素瘤和真菌感染模型。我们的工作将揭开LC的奥秘 具有特定功能的子集，这将为LCS的生物学提供新的见解，并导致 开发基于LC的皮肤病干预策略。