Role of the ultraconserved genomic regions (UCRs) in hematopoiesis

超保守基因组区域 (UCR) 在造血中的作用

• 批准号: 8119543
• 负责人: Ramiro Garzon
• 金额: $ 19.06万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8119543
• 关键词: Binding Sites; Biological Assay; Biology; Blood; Blood Cells; Blood Platelets; Bone Marrow Transplantation; CD34 gene; Cells; Code; Complementary DNA; Complex; Data; Development; Disease; Elements; Erythrocytes; Erythroid; Family; Functional RNA; Gene Expression; Genome; Genomics; Goals; Hematopoiesis; Hematopoietic; Hematopoietic System; Hematopoietic stem cells; Human; Human Genome; Immunophenotyping; In Vitro; Malignant Neoplasms; Megakaryocytes; MicroRNAs; Molecular; Morphology; Names; Oncogenes; Pattern; Play; Process; Regulation; Reporting; Research; Role; Signal Pathway; Staging; System; Tissues; Transcription Initiation Site; Tumor Suppressor Proteins; Work; cell type; human GATA1 protein; in vivo; insight; leukemia; meetings; mouse genome; novel; peripheral blood; promoter; public health relevance; rat genome; research study; stem cell differentiation; transcription factor

DESCRIPTION (provided by applicant): The hematopoietic system is one of the most complex tissues and includes a number of different cell types essential for survival. This process is highly regulated by the complex interplay of signaling pathways and changing sets of transcription factors that are present in hematopoietic cells. The discovery of a new class of transcribed ultraconserved genomic regions (UCRs) that are aberrantly expressed in human leukemias, prompt us to seek whether these UCRs are differentially expressed among the specific hematopoietic lineages and whether UCRs play a role in the differentiation of hematopoietic stem cells (HPCs) into specific lineages, in particular the erythroid and megakaryocytic lineages. Using a novel UCRs microarray platform we have characterized UCRs expression at different stages of hematopoietic differentiation and identified distinctive signatures associated with particular lineages. The overall goal of this proposal is to demonstrate that transcribed UCRs are functionally relevant during hematopoiesis. To achieve this goal we propose the following specific aims: 1) Specific Aim #1: To investigate the effects of ectopic over-expression or inhibition of UCRs expression on erythrocyte and megakaryocyte lineage differentiation "in vitro" and "in vivo" by using morphology, immunophenotype, colony assays and bone marrow transplant assay experiments and 2) Specific Aim #2: To investigate whether GATA-1 regulates UCRs expression by: 1) analyzing UCRs expression after GATA-1 activation using a conditional GATA-1 cell system; 2) identifying UCRs transcription starting sites using Rapid Amplification of cDNA ends (RACE); 3) identifying GATA-1 binding sites using Chip-sequencing and 4) performing promoter functional studies. This research has the potential to identify novel regulators of hematopoiesis and may give insights into basic biology of gene expression and cell fate determination. If successful, this research will uncover an unparalleled reservoir of molecular information on ultraconserved non-coding and coding RNAs in stem cell differentiation that will be extraordinarily valuable for the field. PUBLIC HEALTH RELEVANCE: This research has the potential to give insights into the basic mechanisms of blood formation and to identify novel regulators in this process. This new information will be extraordinarily valuable for the field. Potential benefits include: 1) direct insight into normal hematopoietic regulation will also be important to understand malfunction in leukemias and other hematologic disorders and 2) the pharmacological modulation of hematopoietic stem cells by targeting critical regulators could be used to treat a wide variety of blood cell disorders that arise from abnormalities in the red or platelet family.

描述（由申请人提供）：造血系统是最复杂的组织之一，包括许多生存所必需的不同细胞类型。这一过程受到信号通路和造血细胞中存在的转录因子组变化的复杂相互作用的高度调节。人类白血病中异常表达的一类新的转录超保守基因组区域（UCR）的发现，促使我们寻找这些UCR是否在特定造血谱系中存在差异表达，以及UCR是否在造血干细胞的分化中发挥作用。 （HPC）进入特定谱系，特别是红系和巨核细胞谱系。使用新型 UCR 微阵列平台，我们表征了造血分化不同阶段的 UCR 表达，并鉴定了与特定谱系相关的独特特征。该提案的总体目标是证明转录的 UCR 在造血过程中具有功能相关性。为了实现这一目标，我们提出以下具体目标： 1) 具体目标#1：利用形态学研究异位过度表达或抑制 UCR 表达对“体外”和“体内”红细胞和巨核细胞谱系分化的影响、免疫表型、集落测定和骨髓移植测定实验以及 2) 具体目标 #2：通过以下方式研究 GATA-1 是否调节 UCR 表达：1) 分析后的 UCR 表达使用条件 GATA-1 细胞系统激活 GATA-1； 2) 使用 cDNA 末端快速扩增 (RACE) 识别 UCR 转录起始位点； 3) 使用芯片测序识别 GATA-1 结合位点，4) 进行启动子功能研究。这项研究有可能确定新型造血调节因子，并可能为基因表达和细胞命运决定的基础生物学提供深入的见解。如果成功，这项研究将揭示干细胞分化过程中极其保守的非编码和编码 RNA 的无与伦比的分子信息库，这对该领域来说非常有价值。 公共健康相关性：这项研究有可能深入了解血液形成的基本机制，并确定该过程中的新型调节剂。这些新信息对该领域来说将非常有价值。潜在的好处包括：1）直接了解正常造血调节对于了解白血病和其他血液疾病的功能障碍也很重要；2）通过针对关键调节因子对造血干细胞进行药理调节可用于治疗多种血细胞由红色或血小板家族异常引起的疾病。