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: 9385967
• 负责人: Michelle Leigh Ratliff
• 金额: $ 9.46万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 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 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9385967
• 关键词: ARID3A gene; ATAC-seq; Adult; Affect; Age; Age-Years; Aging; Autoantibodies; Binding; Blood; Blood Cells; Cell Differentiation process; 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; Molecular Profiling; 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; 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 人造血是一个复杂的动态过程，需要对多个基因进行复杂的调节 涉及谱系承诺的途径，导致多种成熟的血细胞类型的发展。 对小鼠和人类的多项研究报告了早期造血的与年龄相关的缺陷，功能缺陷 成熟的老年免疫细胞群体，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。我们实验室的先前研究表明 DNA结合蛋白ARID3A导致造血发育偏离淋巴谱系 祖细胞，表明ARID3A对于正常造血很重要。来自老年人的初步数据 外周血显示出表达ARID3A的造血干细胞（HSC）的频率下降，领先 减少了多能祖细胞和多淋巴祖细胞的数量，但没有变化 与健康的年轻外周血相比，髓样祖细胞的细胞。拟议的研究试图 扩大了体内年轻和衰老的人类外周血造血的初步表型数据 祖细胞的统计显着性，评估和比较年轻ARID3A-的发育潜力 表达体外和体内的年轻ARID3A缺陷和老化的HSC。我们将进一步检查基因 年轻人和老年之间的表达差异以及ARID3A表达和ARID3A缺乏的表达差异 RNA-Seq的HSC。差异表达的基因将进一步分析ARID3A结合和染色质 通过ChIP-Seq和ATAC-Seq分析进行重塑。这些测序技术将是 培训该提案的部分。最后，我们将评估年轻ARID3A的年轻人的能力 ARID3A缺乏和旧的HSC植入了人源化小鼠，以对免疫反应进行免疫反应 接种肺炎链球菌后PC-KLH和生存。拟议研究的总体目标是 确定ARID3A表达在HSC中对免疫衰老和先天免疫反应的影响。 这些研究的完成将根据ARID3A表达对HSC进行分层，识别ARID3A调节的基因 对于衰老很重要，并确定潜在的治疗靶标，以改善老年人的免疫学功能 个人。