Defining the Niche-dependent Role of RNA Editing in Aged and MDS Hematopoietic Stem and Progenitor Cell Dysfunction

定义 RNA 编辑在老年和 MDS 造血干细胞和祖细胞功能障碍中的生态位依赖性作用

• 批准号: 10252784
• 负责人: Catriona Helen Macleod Jamieson
• 金额: $ 35.55万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-20 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10252784
• 关键词: 3' Untranslated Regions; Adenosine; Age; Aging; Biological; Biological Assay; Bone Marrow; Bone Marrow Diseases; Cell Aging; Cell Cycle; Cell Cycle Deregulation; Cell Cycle Kinetics; Cell Cycle Regulation; Cell Lineage; Cell Maintenance; Cells; Coculture Techniques; Code; Cytokine Signaling; DNA; DNA Sequence Alteration; DNA sequencing; Deaminase; Degenerative Disorder; Diagnostic; Double-Stranded RNA; Dysmyelopoietic Syndromes; Endothelial Cells; Enzymes; Event; Evolution; Family; Functional disorder; Gene Expression; Genetic Transcription; Goals; Healthcare; Hematologic Neoplasms; Hematology; Hematopoiesis; Hematopoietic stem cells; Homeostasis; Human; Human Genome; Immunocompromised Host; Impairment; Inflammation; Inflammatory; Inosine; Investigation; Link; Maintenance; Malignant Neoplasms; Mediating; Messenger RNA; MicroRNAs; Mus; Mutation; Myelogenous; Myeloid Progenitor Cells; Pathway interactions; Patients; Play; Population; Prevalence; Primates; Progenitor Cell Engraftment; RNA; RNA Editing; RNA I; RNA Processing; RNA Splicing; Reporter; Research; Resources; Risk; Role; Signal Pathway; Source; Specimen; Tissues; Transcript; Untranslated RNA; Up-Regulation; adenosine deaminase; age related; aged; bone marrow failure syndrome; cdc Genes; cell stroma; cytokine; hematopoietic engraftment; human stem cells; humanized mouse; mouse model; mutant; new therapeutic target; novel therapeutics; premalignant; progenitor; prognostic significance; self-renewal; single-cell RNA sequencing; stem cells; therapy resistant; transcriptome; transcriptome sequencing

Title Defining the Niche-dependent Role of RNA Editing in Aged and MDS Hematopoietic Stem and Progenitor Cell Dysfunction Project Summary During aging, impaired hematopoietic stem and progenitor cell (HSPC) maintenance induced by clonal DNA mutations as well as niche-driven RNA processing deregulation can set the stage for myelodysplastic syndrome (MDS) initiation. Recently, increased adenosine deaminase associated with RNA1 (ADAR1)-mediated A-to-I editing was shown by our group and other research teams to contribute to therapeutic resistance in a broad array of malignancies. Also, we discovered that lentivirally enforced ADAR1 expression in HSPC enhanced myeloid differentiation commensurate with upregulation of PU.1 and reduced dormancy. Whole transcriptome RNA sequencing (RNA-seq) analysis demonstrated that inflammatory cytokine signaling pathways and RNA editing increased during normal aged HSPC evolution to MDS. Thus, we hypothesized that niche dependent activation of RNA editing by ADAR1 provides a competitive advantage for MDS over normal HSPCs. The majority of ADAR1 mediated adenosine-to-inosine (A-to-I) RNA editing events in humans occur within double- stranded RNA (dsRNA) loops created by primate-specific Alu sequences, which comprise 10 percent of the human genome, thereby underscoring that important ADAR1 functional differences exist between human HSPCs compared with their murine counterparts. However, the limited research effort aimed at deciphering the role of ADAR1-mediated RNA editing in HSPC maintenance has been performed primarily in mouse models rather than highly purified human HSPCs. Because ADAR1 is activated by inflammatory cytokines that accelerate aging and MDS initiation, our main goal is to define the niche- dependent role of RNA editing on human HSPC cell fate and cell cycle regulation during aging and MDS initiation. We will first determine the RNA editing profile by whole transcriptome and single cell RNA-seq, RESSqPCR and lentiviral RNA editing reporters. The functional role of ADAR1 in HSPC aging and MDS initiation will be examined in stromal co-cultures with or without addition of inflammatory cytokines, FUCCI2BL cell cycle reporters, and humanized aged HSC and MDS immunocompromised mouse models. We will also examine the effect of RNA editing on APOBEC3 family of DNA deaminase function during MDS initiation. The proposed study is uniquely responsive to PAS-13-033: Stimulating Hematology Investigation: New Endeavors (SHINE) because it will identify the role of ADAR1-mediated regulatory mRNA and miRNA editing in HSPC myeloid lineage commitment and cell cycle deregulation during age-dependent MDS initiation in the inflammatory bone marrow niche. The ultimate goal of this study is to determine the biological, diagnostic and prognostic significance of ADAR1-mediated RNA editing in HSPC aging compared with MDS initiation.

标题定义了RNA编辑在老化和MD的造血词干和 祖细胞功能障碍 项目摘要 在衰老期间，造血茎和祖细胞（HSPC）维持受损。 克隆DNA突变以及小裂驱动的RNA处理放松管制可以为 骨髓增生综合征（MDS）的起始。最近，增加了与 我们的小组和其他研究团队向RNA1（ADAR1）介导的A到I编辑显示了 在广泛的恶性肿瘤中有助于治疗性抗性。另外，我们发现 HSPC中的慢病毒强化ADAR1表达增强了髓样分化与 PU.1的上调和休眠减少。整个转录组RNA测序（RNA-Seq） 分析表明，炎性细胞因子信号通路和RNA编辑增加 在正常老化的HSPC演变为MDS期间。因此，我们假设利基依赖性激活 ADAR1编辑的RNA编辑为MDS比正常HSPC提供了竞争优势。多数 ADAR1介导的腺苷对 - 肌苷（A-to-I）RNA编辑事件发生在双重 由灵长类动物特异性ALU序列创建的滞留的RNA（DSRNA）环，该序列占10％ 人类基因组，从而强调了重要的ADAR1功能差异 人类HSPC与他们的鼠相比。但是，有限的研究工作旨在 已经执行了ADAR1介导的RNA编辑在HSPC维护中的作用 主要在小鼠模型中而不是高度纯化的人类HSPC中。因为adar1被激活 加速衰老和MDS启动的炎性细胞因子，我们的主要目标是定义利基市场 RNA编辑在衰老期间对人HSPC细胞命运和细胞周期调节的依赖性作用 MDS启动。我们将首先通过整个转录组和单个单元格确定RNA编辑曲线 RNA-Seq，RESSQPCR和慢病毒RNA编辑记者。 ADAR1在HSPC中的功能作用 衰老和MDS的启动将在基质共培养中检查，或者不增加或不增加 炎性细胞因子，FucCI2BL细胞周期报告基因以及人源化的老年HSC和MDS 免疫功能低下的小鼠模型。我们还将检查RNA编辑对APOBEC3的影响 MDS启动过程中DNA脱氨酶功能的家族。拟议的研究对 PAS-13-033：刺激血液学调查：新的努力（Shine），因为它将 确定ADAR1介导的调节mRNA和miRNA编辑在HSPC髓样谱系中的作用 在炎症骨中依赖年龄依赖的MDS启动期间的承诺和细胞周期放松管制 骨髓利基。这项研究的最终目标是确定生物学，诊断和预后 与MDS启动相比，HSPC衰老中ADAR1介导的RNA编辑的重要性。