Translational genomics in gout: From GWAS signal to mechanism

痛风的转化基因组学：从 GWAS 信号到机制

• 批准号: 10735151
• 负责人: TONY R MERRIMAN
• 金额: $ 60.56万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-21 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10735151
• 关键词: Acute; Address; Adult; Affect; African American; Alabama; Ankylosing spondylitis; Architecture; Area; Biological Assay; CTLA4-Ig; Candidate Disease Gene; Cardiometabolic Disease; Cell Line; Cells; Chromosome Structures; Chromosomes, Human, 6-12 and X; Clinical; Code; Consent; DNA Methylation; Deposition; Development; Drug Targeting; Enhancers; Epigenetic Process; Family; Flare; Future; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Processes; Genetic Transcription; Genetic study; Genomics; Genotype; Gout; Health; Hematopoiesis; Hi-C; Human; Human Cell Line; Hyperuricemia; Immune; Immune response; Immunity; Individual; Inflammasome; Inflammation; Inflammatory; Inflammatory Response; Injections; Innate Immune System; International; Joints; Knock-out; Knowledge; Larva; Leukocytes; Link; Macrophage; Mediating; Metabolism; Methylation; Modeling; Modification; Molecular; Molecular Conformation; Molecular Genetics; Nucleic Acid Regulatory Sequences; Participant; Pathway interactions; Persons; Population; Predisposition; Process; Production; Proteins; RNA Binding; Regulation; Research; Research Design; Resources; Rheumatoid Arthritis; Role; Secondary to; Severities; Signal Transduction; Site; Stimulus; System; Testing; Therapeutic; Training; Translating; Translations; Untranslated RNA; Urate; Variant; Work; Zebrafish; candidate identification; caucasian American; cell type; chromatin isolation by RNA purification sequencing; comorbidity; drug development; druggable target; epigenome; epigenomics; experience; follow-up; gene conservation; genetic association; genetic variant; genome wide association study; genomic locus; histone modification; immunoregulation; improved; innovation; insight; knock-down; locked nucleic acid; monocyte; neutrophil; new therapeutic target; novel; novel strategies; overexpression; physical symptom; prevent; promoter; psychologic; recruit; response; risk variant; screening; social; targeted treatment; transcriptome; transcriptome sequencing; translational genomics; validation studies; volunteer

SUMMARY Gout is prevalent in the US (3.9% of the adult population) and not only directly impacts peoples’ lives but is also co-morbid with cardiometabolic disease. Gout consists of unpredictable episodes of acute inflammation or flares resulting from monocyte NLRP3 inflammasome activation by monosodium urate (MSU) crystals in people with hyperuricemia, the subsequent production of IL-1, and recruitment of large numbers of inflammatory cells into the affected joint. Epigenetic reprogramming and altered gene transcription in response to elevated levels of soluble urate are mechanisms of this enhanced cellular reactivity to the secondary stimuli, known as innate trained immunity. To better understand the factors controlling the progression from hyperuricemia to gout we have completed a very large genome-wide association study (GWAS) in gout, identifying several hundred gout-associated loci. Loci include long-noncoding RNAs (lncRNAs) that have diverse functions including regulation of gene expression, deposition of epigenetic modifications and organization of chromosome architecture. Some are immune gene-priming lncRNAs (IPLs) that direct transcriptional machinery over multiple promoters. From these loci we have identified one key causal pathway (clonal hematopoiesis of indeterminate potential (CHIP)) for which we will study the molecular genetic processes controlling its activation. The CHIP pathway is involved in control of the epigenome - DNA methylation, histone modification and metabolism of substrates. It may make people more susceptible to gout by causing the innate immune system to be hyper-responsive to MSU crystals. We hypothesize that the genetic loci identified from the gout GWAS control activity of this pathway by regulation of gene expression, including through regulation of the epigenome. In testing this hypothesis, in three Aims, we will understand the molecular control of the pathway and provide a basis for future studies in targeting this pathway for improved management of gout. In Aim 1 we will use experimental systems, including a zebrafish model of gout, to understand where regulatory regions are expressed and how the gout-associated loci influence NLRP3- inflamasome activation. Zebrafish are translucent, allowing development of innovative models, and there is 82% conservation of genes with humans and functionally equivalent macrophages and neutrophils. In Aim 2 we will compare the transcriptome and epigenome in MSU crystal-stimulated monocytes of individuals with high and low burdens of gout risk alleles in order to gain further insights into the molecular regulation of the pathways, and to identify downstream pathways. Individual genes will be knocked down in a cell line. In Aim 3 we will use our established pipeline to understand how lncRNAs (i.e. IPLs) connect transcriptional machinery to the promoters of innate immune genes at specific loci in the molecular control of activation of the gout flare. Our studies will deepen our knowledge of the mechanisms of gout flares and its genetic basis, and ultimately point to areas of research that may allow for novel treatments in gout.

概括 痛风在美国普遍存在（占成年人口的3.9％），不仅会直接影响人们的生活 也与心脏代谢疾病合并。痛风由不可预测的急性炎症或 单核细胞NLRP3炎性菌株（MSU）晶体激活产生的耀斑 患有高尿酸血症的人，随后的IL-1以及大量的招募 炎症细胞进入受影响的关节。表观遗传重编程和改变的基因转录反应 达到固体尿酸盐水平是这种增强的细胞反应性的机制， 被称为先天训练的免疫力。更好地了解控制从 高尿液症，我们已经完成了一项大型全基因组协会研究（GWAS）， 识别数百个相关的基因座。基因座包括具有长期编码的RNA（LNCRNA） 潜水功能，包括调节基因表达，表观遗传修饰的沉积和 染色体体系结构的组织。有些是导演 多个启动子上的转录机械。在这些基因座上，我们已经确定了一个关键因果途径 （不确定电势的克隆造血（芯片）），我们将研究分子遗传 处理控制其激活的过程。芯片途径与表观基因组-DNA的控制有关 甲基化的甲基化，组蛋白的修饰和代谢。它可能会使人们更容易获得 通过导致先天免疫系统对MSU晶体具有超反应性。我们假设 通过调节基因表达的调节，从该途径的痛风GWAS控制活性定位的遗传 包括通过调节表观基因组。在检验这一假设时，我们将了解 途径的分子控制，为以后的研究提供了针对此途径的改进的基础 痛风的管理。在AIM 1中，我们将使用实验系统（包括斑马模型痛风模型） 了解监管区域的表达位置以及与GOUT相关的基因座如何影响NLRP3- 激活。斑马鱼是半透明的，可以开发创新的模型，并且有 82％的基因和人类和功能等效的巨噬细胞和中性粒细胞保存。在目标2中 我们将比较MSU晶体刺激的单核细胞中的转录组和表观基因组 高和低燃烧的GET风险等位基因，以进一步了解分子调节 途径，并识别下游途径。单个基因将被击倒在细胞系中。在目标3中 我们将使用我们既定的管道来了解LNCRNA（即IPL）如何连接转录机械 在特定局部的先天免疫基因的启动子中，在痛风耀斑激活的分子控制中。 我们的研究将加深我们对痛风耀斑机制及其遗传基础的了解，并最终 指向研究领域，可以在痛风中进行新的治疗方法。