Mechanistic modeling of epigenetic modifier mutations in human pluripotent stem cell-derived immune cells

人类多能干细胞衍生的免疫细胞表观遗传修饰突变的机制模型

• 批准号: 10733331
• 负责人: Minji Byun
• 金额: $ 19.63万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10733331
• 关键词: ATAC-seq; Ablation; Acceleration; Address; Affect; Age; Aging; Atherosclerosis; Automobile Driving; Base Pairing; Binding; Biochemical; Biological; Biology; Blood Cells; Cardiovascular Diseases; Cell model; Cells; ChIP-seq; Chromatin; Clinical; Clonal Expansion; Clone Cells; Complex; DNA Methylation; DNMT3a; Data; Defect; Elderly; Enhancers; Enzymes; Epigenetic Process; Experimental Models; Family; Future; Gene Combinations; Gene Expression; Gene Expression Profiling; Genes; Genetic Diseases; Genetic Models; Genetic Transcription; Hematologic Neoplasms; Hematology; Hematopoiesis; Heterozygote; Human; Immune; Immune response; Impairment; Individual; Inflammation; Knowledge; Macrophage; Measurement; Measures; Modeling; Molecular; Morbidity - disease rate; Mus; Mutate; Mutation; Myelogenous; Myeloid Cells; Null Lymphocytes; Orthologous Gene; Outcome; Pathogenicity; Pathology; Pathway interactions; Phenotype; Population; Prevention; Proteins; Research; Research Personnel; Resolution; Risk; Side; Somatic Mutation; Testing; Work; atherosclerosis risk; demethylation; differential expression; genetic manipulation; human pluripotent stem cell; in vitro Model; in vivo; knock-down; loss of function; loss of function mutation; mortality; public health priorities; therapeutic target; transcription factor; transcriptome

PROJECT SUMMARY Clonal hematopoiesis of indeterminate potential (CHIP) refers to the presence of expanded blood cell clones with one or more somatic mutations without other hematologic abnormalities. CHIP is common in the elderly, affecting more than 10% of individuals over 65 years. CHIP is associated with a 10-fold increase risk of hematologic malignancies and a doubled risk of atherosclerotic cardiovascular disease, contributing to an increase in all-cause mortality. The two most commonly mutated genes in CHIP are DNMT3A and TET2, both of which encode epigenetic modifiers. Recent studies have found evidence of increased inflammation and worsened atherosclerosis when the murine ortholog of TET2 was perturbed in myeloid cells in vivo. However, the specific epigenetic mechanism underlying this phenotype and whether DNMT3A and TET2 mutations target the same biological pathway are unclear. Furthermore, it remains unexplained why loss-of-function mutations in DNMT3A and TET2 have shared a clinical outcome despite the two genes encode enzymes with opposite biochemical functions (DNA methylation vs. de-methylation). To address this knowledge gap, we developed human pluripotent stem cell (hPSC)-derived macrophage models of DNMT3A- and TET2- haploinsufficiency. hPSC-derived macrophages are inexhaustible, scalable, and amenable to genetic manipulation, offering a powerful in vitro model well-suited for mechanistic studies. In this proposal, we propose the use of this model to (1) define unique and shared epigenetic features of DNMT3A and TET2 haploinsufficiency, and (2) identify transcription factors driving altered immune gene expression in DNMT3A- and TET2-haploinsufficient immune cells. This project is built on our previous work, and we anticipate that findings from this study will guide future studies on targeting an epigenetic vulnerability shared between DNMT3A- and TET2-mutated cells as well as mechanistic studies on other drivers of CHIP.

项目摘要 不确定电势（CHIP）的克隆造血是指扩张的血细胞克隆的存在 具有一个或多个没有其他血液学异常的体细胞突变。芯片在老年人中很常见， 在65年内影响超过10％的个体。芯片与增加10倍的风险有关 血液系统恶性肿瘤和动脉粥样硬化心血管疾病的两倍风险，有助于 全因死亡率的增加。芯片中两个最常见的突变基因是DNMT3A和TET2，都 其中编码表观遗传修饰符。最近的研究发现了炎症增加和 当TET2的鼠直系同源物在体内扰动TET2的鼠直系同源物时，动脉粥样硬化恶化。然而， 该表型的基础的特定表观遗传机制以及DNMT3A和TET2突变是否 目标尚不清楚相同的生物途径。此外，仍然无法解释为什么功能丧失 尽管有两个基因编码酶，但DNMT3A和TET2中的突变仍具有临床结果 相反的生化功能（DNA甲基化与脱甲基化）。为了解决这个知识差距，我们 开发了人类多能干细胞（HPSC）衍生的DNMT3A和TET2-的巨噬细胞模型 单倍不足。 HPSC衍生的巨噬细胞是取之不尽的，可扩展的，并且可与遗传 操纵，提供强大的体外模型，非常适合机械研究。在这个建议中，我们 提出该模型的使用来定义DNMT3A和TET2的独特和共享的表观遗传特征 单倍不足，（2）确定转录因子驱动DNMT3A-中的免疫基因表达改变的转录因子 和TET2 - 悬子免疫细胞。该项目建立在我们以前的工作上，我们预计 这项研究的发现将指导未来针对针对表观遗传脆弱性的研究 DNMT3A和TET2突变的细胞以及对其他芯片驱动因素的机理研究。