Investigation of Epigenetic Dysregulation in Lupus NK Cells

狼疮 NK 细胞表观遗传失调的研究

• 批准号: 10363743
• 负责人: PAUL JOSEPH UTZ
• 金额: $ 19.68万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-03 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10363743
• 关键词: Affect; Antigen-Antibody Complex; Atlases; Benchmarking; Biological; Biological Assay; Blood donor; Cell Adhesion Molecules; Cell Compartmentation; Cell physiology; Cell surface; Cell-Mediated Cytolysis; Cells; Cellular biology; ChIP-seq; Characteristics; Chromatin; Clinical; Clinical Research; Complex; Cytometry; DNA Methylation; DNA Sequence Alteration; Data; Data Set; Development; Disease; Epigenetic Process; Fluorescence-Activated Cell Sorting; Foundations; Frequencies; Functional disorder; Future; Genetic; Genomics; Goals; Granzyme; Heterogeneity; Histone Acetylation; Histone Deacetylase; Histones; Immune; Immune Tolerance; Immune response; Immune system; Individual; Interferon Type II; Intervention; Investigation; Light; Link; Lupus; Lysine; MHC Class I Genes; Measures; Mediating; Methods; Modeling; Molecular; Monitor; Monozygotic twins; NK Cell Activation; Natural Killer Cells; Other Genetics; Outcome; Pathologic; Patients; Pattern; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Pharmacology; Phenotype; Play; Population; Production; Proteomics; Receptor Cell; Refractory; Regulation; Resolution; Role; Sampling; Systemic Lupus Erythematosus; TNF gene; Technology; Testing; Therapeutic Intervention; Transposase; Virus Diseases; Vorinostat; antibody-dependent cell cytotoxicity; base; clinically significant; cytokine; cytotoxicity; disease heterogeneity; disorder risk; epigenetic marker; epigenetic variation; epigenomics; genomic locus; healthy volunteer; high dimensionality; high reward; high risk; histone modification; human disease; immune function; indexing; innovation; insight; novel; novel therapeutics; perforin; peripheral blood; receptor; single cell analysis; single cell technology; systemic inflammatory response

PROJECT SUMMARY The overarching goal of this proposal is to systematically characterize an epigenetically distinct natural killer (NK) cell population with hypoacetylated histones. This NK cell subset is reduced in frequency in systemic lupus erythematosus (SLE) patients with increased disease activity. We will test the hypothesis that this NK cell subset, which we term “NKSLE cells”, has a distinct cell surface molecule profile, and is functionally different from other NK cells in cytolytic capacity and cytokine secretion upon activation. We will further examine if NK cell functions can be altered by experimental manipulation of histone acetylation. Our data will lay a strong foundation for future epigenomic studies on specific genes or genomic loci pivotal for lupus pathophysiology. The advent of single-cell technologies has greatly accelerated our understanding of immune system heterogeneity. Increasing evidence supports a compelling model in which pathologically important cells in SLE may represent only a small fraction of the highly complex immune system. This proposal builds upon this innovative concept, and will leverage cutting-edge technologies to interrogate the epigenetic and proteomic heterogeneity of NK cells, and their complex relationships. In Aim 1, we will employ highly multiplexed mass cytometry to monitor a broad array of functionally important NK cell markers, such as activating and inhibitory receptors, cell adhesion molecules, and those indicative of cytotoxicity activity, and histone acetylation abundance in individual NK cells. We will first benchmark NK cell heterogeneity in healthy blood donors, with a goal to identify cellular features associated with global histone acetylation level. Next, we will extend the analysis to SLE patients, whose NK cell compartments show reduced representation of cells with hypoacetylated histones. This study will reveal the phenotypic and functional characteristics of NKSLE cells. In Aim 2, we will investigate the functional differences between NK cells with differential histone acetylation content. Cytotoxicity and proinflammatory cytokine secretion are key immunological functions mediated by NK cells during a viral infection. Mass cytometry will be utilized to examine how differential histone acetylation abundance in individual NK cells is correlated with the degree of their immune response upon activation measured by degranulation markers, and cytokine production. Our analysis will first focus on NK cells isolated from healthy volunteers, and subsequently extend to those from lupus patients to better understand the clinical significance of NKSLE cell reduction in lupus. Lastly, as opposed to genetic mutations that are generally refractory to therapeutic interventions, epigenetic phenomena are largely reversible, making them attractive targets for developing new drugs. We will investigate the plasticity of NK cell functions by pharmacologically manipulating histone acetylation. Together, our proposal will provide new insights into the biological and pathological significance of histone acetylation in NK cell molecular and functional phenotypes. Our data will be valuable for future studies on lupus and other human diseases in which NK cells play important pathological roles.

项目摘要 这项建议的首要目标是系统地描述一个表观遗传学上不同的自然 具有低乙酰化组蛋白的杀伤（NK）细胞群。这种NK细胞亚群在全身性免疫中的频率降低， 红斑狼疮（SLE）患者的疾病活动性增加。我们将检验NK细胞 我们称之为“NKSLE细胞”的亚群具有不同的细胞表面分子谱，并且在功能上不同于 其他NK细胞在活化时的细胞溶解能力和细胞因子分泌。我们将进一步检查NK细胞 功能可以通过组蛋白乙酰化的实验操作来改变。我们的数据将奠定坚实的基础 用于将来对狼疮病理生理学关键的特定基因或基因组位点的表观基因组学研究。的出现 单细胞技术大大加速了我们对免疫系统异质性的理解。增加 证据支持一个令人信服的模型，其中SLE中病理重要的细胞可能仅代表一小部分 高度复杂的免疫系统的一部分。这一建议建立在这一创新概念的基础上， 利用尖端技术来研究NK细胞的表观遗传和蛋白质组异质性， 复杂的关系。在目标1中，我们将采用高度多路复用的质谱细胞术来监测广泛的阵列 功能重要的NK细胞标志物，如活化和抑制受体，细胞粘附分子， 以及指示细胞毒性活性和单个NK细胞中组蛋白乙酰化丰度的那些。我们将首先 健康献血者中的基准NK细胞异质性，目的是识别与以下相关的细胞特征： 整体组蛋白乙酰化水平。接下来，我们将分析扩展到SLE患者，其NK细胞区室 显示具有低乙酰化组蛋白的细胞的代表性降低。这项研究将揭示表型和 NKSLE细胞的功能特征。在目标2中，我们将研究NK细胞之间的功能差异， 组蛋白乙酰化含量不同。细胞毒性和促炎细胞因子分泌是关键 病毒感染期间NK细胞介导的免疫功能。将使用质谱细胞仪检查 单个NK细胞中差异组蛋白乙酰化丰度如何与其免疫反应程度相关 通过脱粒标志物测量的活化后的反应和细胞因子产生。我们的分析将首先 专注于从健康志愿者中分离的NK细胞，随后扩展到狼疮患者， 更好地理解狼疮中NKSLE细胞减少的临床意义。最后，相对于基因 通常对治疗干预无效的突变，表观遗传现象在很大程度上是可逆的， 使它们成为开发新药的有吸引力的目标。我们将研究NK细胞功能的可塑性， 通过人工操纵组蛋白乙酰化。总之，我们的提案将提供新的见解， 组蛋白乙酰化在NK细胞分子和功能表型中的生物学和病理学意义。 我们的数据将是有价值的未来研究狼疮和其他人类疾病中的NK细胞发挥重要作用 病理角色