Role of ARID3a in aging human hematopoietic progenitor cells

ARID3a在人类造血祖细胞衰老中的作用

• 批准号: 10263295
• 负责人: Michelle Leigh Ratliff
• 金额: $ 24.49万
• 依托单位: EAST CAROLINA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10263295
• 关键词: ARID3A gene; ATAC-seq; Adult; Affect; Age; Age-Years; Aging; Autoantibodies; Binding; Blood; Blood Cells; Cells; ChIP-seq; Complex; DNA-Binding Proteins; Data; Defect; Development; Dominant-Negative Mutation; Elderly; Epigenetic Process; European; Exhibits; Flow Cytometry; Frequencies; Future; Gene Expression; Gene Expression Regulation; Genes; Goals; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; Human; Immune; Immune response; Immunization; Impairment; In Vitro; Individual; Influenza; Innate Immune Response; Keyhole Limpet Hemocyanin; Knockout Mice; Lead; Lymphoid; Multipotent Stem Cells; Mus; Myelogenous; Myeloid Progenitor Cells; Pathway interactions; Patients; Play; Population; Population Study; Process; Production; Regimen; Regulation; Reporting; Role; Streptococcus pneumoniae; Systemic Lupus Erythematosus; Techniques; Training; Transgenic Mice; Vaccination; age related; aged; base; cell type; chromatin remodeling; differential expression; hematopoietic stem cell differentiation; hematopoietic stem cell expansion; human subject; humanized mouse; immune function; immunosenescence; improved; in vivo; in vivo Model; insight; new therapeutic target; overexpression; pathogen; peripheral blood; phenotypic data; progenitor; response; self-renewal; senescence; small hairpin RNA; therapeutic target; transcription factor; transcriptome; transcriptome sequencing

Project Summary Human hematopoiesis is a complex, dynamic process that requires intricate regulation of multiple gene pathways involved in lineage commitments, leading to development of multiple mature blood cell types. Multiple studies in mice and humans report age-related defects in early hematopoiesis and functions defects in the mature aged immune cell populations, which leads to impaired immune responses to pathogens. These defects are especially important because population studies estimate that nearly 40% of European and US populations will be over 60 years of age by 2050. Previous studies in our lab indicate that loss of expression of the DNA-binding protein ARID3a leads to hematopoietic developmental skewing away from lymphoid lineage progenitors, indicating that ARID3a is important for normal hematopoiesis. Preliminary data from aged human peripheral blood shows decreased frequency of ARID3a-expressing hematopoietic stem cells (HSCs), leading to decreased numbers of multipotent progenitors and multi-lymphoid progenitors, but no alterations in numbers of myeloid progenitor cells when compared to healthy young peripheral blood. The proposed studies seek to expanded the preliminary phenotypical data of ex vivo young and aged human peripheral blood hematopoietic progenitor cells for statistical significance, evaluate and compare developmental potential of young ARID3a- expressing, young ARID3a-deficient and aged HSCs in vitro and in vivo. We will further examine the gene expression differences between young and old, as well as between ARID3a expressing and ARID3a deficient HSCs by RNA-seq. Differentially expressed genes will be further analyzed for ARID3a binding and chromatin remodeling by ChIP-seq and ATAC-seq analyses. These sequencing techniques will be the focus of the training portion of this proposal. Finally, we will evaluate the capacity of young ARID3a-expressing, young ARID3a-deficient, and old HSC engrafted humanized mice to mount immune responses to immunization with PC-KLH and survival following inoculation with S. pneumoniae. The overall goal of the proposed studies is to determine the effect of ARID3a expression in HSCs on immunosenescence and innate immune responses. Completion of these studies will stratify HSCs based on ARID3a expression, identify ARID3a-regulated genes important for aging, and identify potential therapeutic targets for improving immunological function in elderly individuals.

项目摘要 人类造血是一个复杂的动态过程，需要多个基因的复杂调控 参与谱系承诺的途径，导致多种成熟血细胞类型的发展。 在小鼠和人类中进行的多项研究报告了与年龄相关的早期造血缺陷和功能缺陷， 成熟老化的免疫细胞群，这导致对病原体的免疫反应受损。这些 缺陷尤其重要，因为人口研究估计，近40%的欧洲和美国 到2050年，人口将超过60岁。我们实验室以前的研究表明， DNA结合蛋白ARID 3a导致造血发育偏离淋巴系 这表明ARID 3a对于正常造血是重要的。老年人的初步数据 外周血显示表达ARID 3a的造血干细胞（HSC）的频率降低， 多能祖细胞和多淋巴祖细胞数量减少，但数量无变化 与健康年轻人外周血相比，拟议的研究旨在 扩大了体外年轻和老年人外周血造血干细胞的初步表型数据， 祖细胞的统计学意义，评估和比较年轻的ARID 3a- 在体外和体内表达年轻的ARID 3a缺陷的和老化的HSC。我们将进一步研究 年轻人和老年人之间以及ARID 3a表达和ARID 3a缺陷之间的表达差异 通过RNA-seq.将进一步分析差异表达基因的ARID 3a结合和染色质 通过ChIP-seq和ATAC-seq分析进行重构。这些测序技术将是本领域的重点。 这一建议的培训部分。最后，我们将评估年轻的ARID 3a表达的能力，年轻的 ARID 3a缺陷和年老HSC移植的人源化小鼠，以产生对免疫的免疫应答， PC-KLH和接种S.肺炎。拟议研究的总体目标是 确定HSC中ARID 3a表达对免疫衰老和先天免疫应答的影响。 这些研究的完成将基于ARID 3a表达对HSC进行分层，鉴定ARID 3a调控的基因， 重要的老化，并确定潜在的治疗靶点，以改善老年人的免疫功能 个体