Investigation of Epigenetic Dysregulation in Lupus NK Cells

狼疮 NK 细胞表观遗传失调的研究

基本信息

批准号：
10194280
负责人：
PAUL JOSEPH UTZ
金额：
$ 23.63万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-03 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10194280
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

项目摘要

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.

项目摘要该提案的总体目标是系统地表征表观遗传上不同的自然杀手级（NK）细胞群含有低乙酰化组蛋白。该NK细胞子集的全身性频率降低狼疮性红斑患者（SLE）患者疾病活性增加。我们将测试这个NK细胞的假设我们称其为“ nksle细胞”的子集具有不同的细胞表面分子曲线，并且在功能上不同激活后，其他NK细胞以细胞溶解能力和细胞因子分泌。我们将进一步检查NK细胞是否通过对组蛋白乙酰化的实验操纵可以改变功能。我们的数据将奠定坚实的基础对于未来的表观基因组学研究，有关狼疮病理生理学的特定基因或基因组基因组关键。冒险单细胞技术极大地加速了我们对免疫系统异质性的理解。增加证据支持一个引人注目的模型，在该模型中，SLE中重要的重要细胞可能仅代表一个小的高度复杂的免疫系统的一部分。该提议建立在这个创新的概念的基础上，并将利用尖端技术来询问NK细胞的表观遗传和蛋白质组学异质性，以及他们复杂的关系。在AIM 1中，我们将采用高度多路复用的质量细胞仪来监视宽阵列功能上重要的NK细胞标记物（例如激活和抑制受体），细胞粘附分子，以及指示细胞毒性活性以及各个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的基准NK细胞异质性，目的是识别与细胞特征相关的细胞特征全球组蛋白乙酰化水平。接下来，我们将将分析扩展到SLE患者，其NK细胞室的患者表现出降低的用乙酰化组蛋白的细胞表示。这项研究将揭示表型和 NKSLE细胞的功能特征。在AIM 2中，我们将研究NK细胞之间的功能差异具有差异性组蛋白乙酰化含量。细胞毒性和促炎性细胞因子分泌是关键 NK细胞在病毒感染过程中介导的免疫功能。质量细胞仪将用于检查单个NK细胞中的差异组蛋白乙酰化丰度如何与免疫的程度相关通过脱粒标记和细胞因子产生测量激活后的反应。我们的分析将首先专注于从健康志愿者中分离出的NK细胞，随后扩展到狼疮患者到更好地了解狼疮中NKSLE细胞降低的临床意义。最后，与遗传相反通常对热干预的突变，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药物操纵组蛋白乙酰化。一起，我们的建议将为您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。我们的数据对于将来对NK细胞发挥重要作用的狼疮和其他人类疾病的研究很有价值病理角色。