Altered inflammatory response associated with acquired DNMT3A mutations

与获得性 DNMT3A 突变相关的炎症反应改变

• 批准号: 10746603
• 负责人: Minji Byun
• 金额: $ 66.53万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10746603
• 关键词: Address; Affect; Age; Animal Model; Base Pairing; Binding; Biological Assay; Blood Cells; Cardiovascular Diseases; Cell Differentiation process; Cell Lineage; Cell model; Cells; Chromatin; Chromatin Remodeling Factor; Clonal Expansion; Computerized Medical Record; Coronary heart disease; Coupled; DNA; DNA Methylation; DNMT3a; DNMT3a mutation; Data; Defect; Development; Diagnosis; Disease; Disease model; Elderly; Enrollment; Enzymes; Epigenetic Process; Functional disorder; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Genotype; Goals; Hematologic Neoplasms; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; Heterozygote; Human; IL6 gene; Immunology; Individual; Inflammation; Inflammatory; Inflammatory Response; Interleukin-6; Knowledge; Link; Macrophage; Malignant Neoplasms; Mediating; Modeling; Molecular; Molecular Profiling; Mus; Mutate; Mutation; Mutation Analysis; Myeloid Cells; Myelopoiesis; Nucleic Acid Regulatory Sequences; Orthologous Gene; Output; Pathologic; Patients; Phenotype; Population Heterogeneity; Positioning Attribute; Predisposition; Prevention approach; Proteins; Proteomics; Public Health; Regulator Genes; Repressor Proteins; Research Personnel; Risk; Role; Source; Study models; System; Techniques; Technology; United States; aged; aging population; biobank; cell type; chromatin remodeling; cohort; cytokine; design; drug use screening; epigenetic drug; ethnic diversity; genetic manipulation; genome editing; genome-wide; genomic locus; human embryonic stem cell; human pluripotent stem cell; induced pluripotent stem cell; insight; monocyte; mutation carrier; neutrophil; novel; novel strategies; novel therapeutic intervention; null mutation; prevent; promoter; recruit; single-cell RNA sequencing; stem cell biology; stem cell model; transcription factor; transcriptome sequencing; transcriptomics

PROJECT SUMMARY The human gene DNMT3A encodes one of the three enzymes that carry out DNA methylation in humans. Clonal expansion of blood cells with acquired mutations in DNMT3A is common in older adults, occurring in 5-10 % of healthy individuals aged 60 or above. Carriers of DNMT3A mutations have an approximately tenfold increased risk of developing hematologic cancers and are twice as likely to develop coronary heart disease. Given the rapidly aging population in the United States and worldwide, understanding the mechanistic basis of the association between acquired DNMT3A mutations in blood cells and increased susceptibility to cancer and cardiovascular disease is tremendously important for public health. Recent studies found evidence of increased inflammation mediated by myeloid cells when the ortholog of DNMT3A was perturbed in animal models. However, the molecular mechanisms underlying this phenomenon and whether DNMT3A mutations affect the inflammatory response of human myeloid cells remain poorly understood. To address this gap in knowledge, we established an experimental system based on myeloid cells differentiated from human pluripotent stem cells. Using this system, we found that human macrophages with DNMT3A mutations displayed altered inflammatory response compared to wild-type macrophages, characterized by augmented expression of IL-6, a potent proinflammatory cytokine. The IL6 promoter was one of the most significantly hypomethylated loci in DNMT3A-mutated macrophages, suggesting a direct mechanistic link between DNA methylation and inflammatory response in our model. In this application, we propose to characterize the molecular signature of the inflammatory response associated with DNMT3A mutations using genetically defined human macrophages and neutrophils and to dissect the epigenetic mechanisms underlying DNMT3A-mediated gene expression regulation. In addition, we will examine the impact of harboring clonally expanded blood cells with DNMT3A mutations on the inflammatory response of primary myeloid cells using a novel single-cell transcriptomic technique. We are in an ideal position to pursue this project given the availability of human pluripotent stem cell-based human myeloid cell models that we have developed and validated, our access to a large biobank representing extremely diverse populations, and the assembly of a strong scientific team consisting of investigators with complementary expertise. Findings from the proposed study will provide critical new insights into the consequence of acquiring DNMT3A mutations on inflammation, and help us develop novel strategies to prevent and treat pathologic conditions related to DNMT3A mutations.

项目摘要 人类基因DNMT3A编码在三种中进行DNA甲基化的酶之一 人类。在DNMT3A中获得的突变的血细胞的克隆扩张在老年人中很常见， 发生在5-10％的60岁或更高的健康个体中。 DNMT3A突变的载体具有 大约增加了患血液学癌症的风险大约增加了，并且可能发展的可能性是 冠心病。鉴于美国和全球的人口迅速衰老，理解 在血细胞中获得的DNMT3A突变与增加的机械基础 对癌症和心血管疾病的敏感性对于公共卫生非常重要。最近的研究 发现DNMT3A的直系同源 动物模型受到干扰。但是，这种现象的基础机制以及是否 DNMT3A突变会影响人髓样细胞的炎症反应仍然很少理解。到 在知识上解决这一差距，我们建立了一个基于髓样细胞的实验系统 来自人类多能干细胞。使用此系统，我们发现具有DNMT3A的人类巨噬细胞 与野生型巨噬细胞相比，突变显示出改变的炎症反应，其特征是 增强IL-6的表达，这是一种有效的促炎细胞因子。 IL6启动子是最多的 在DNMT3A突变的巨噬细胞中显着降低了甲基化的基因座，这表明了直接机械链接 在我们的模型中DNA甲基化和炎症反应之间。在此应用程序中，我们建议 表征与DNMT3A突变相关的炎症反应的分子特征 遗传定义的人类巨噬细胞和中性粒细胞，并剖析了依据的表观遗传机制 DNMT3A介导的基因表达调节。此外，我们将研究克隆藏身的影响 使用A的原代髓样细胞的炎症反应扩大血细胞，使用A 新型的单细胞转录组技术。我们处于理想的位置，可以在 我们已经开发的人类多能干细胞的人类髓样细胞模型的可用性 经过验证，我们进入代表极度多样化人群的大型生物库的访问，以及 强大的科学团队由具有互补专业知识的研究人员组成。提议的发现 研究将为获取DNMT3A突变对炎症的结果提供关键的新见解， 并帮助我们制定新的策略，以预防和治疗与DNMT3A突变有关的病理状况。