Role of C/EBPepsilon in myeloid differentiation

C/EBPepsilon 在骨髓分化中的作用

• 批准号: 7188117
• 负责人: Stephanie Halene
• 金额: $ 12.85万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7188117
• 关键词: Affect; Appearance; Back; Biological Assay; Bone Marrow; Bone Marrow Transplantation; Candidate Disease Gene; Cell Line; Cell Nucleus; Cells; Characteristics; Chemotaxis; Cytoplasmic Granules; Defect; Disruption; Estrogen Receptors; Event; Failure; Gelatinase A; Gene Expression; Gene Proteins; Genes; Hematopoietic stem cells; Infection; Injection of therapeutic agent; Knockout Mice; Lactoferrin; Marrow; Microarray Analysis; Modeling; Mus; Myelocyte; Myelodysplastic/Myeloproliferative Disease; Myelogenous; Myeloproliferation; Neutrophil Collagenase; Patients; Phagocytosis; Phenotype; Play; Polymerase Chain Reaction; Process; Progranulocytes; Proteins; Reporting; Respiratory Burst; Retroviral Vector; Role; SCID Mice; Series; Site; Staging; Transcobalamin (I); Transcriptional Regulation; Transducers; Transplantation; Up-Regulation; bactericide; established cell line; granulocyte; in vivo; insight; leukemia; neutrophil; progenitor; programs; protein expression; retroviral transduction; transcription factor

DESCRIPTION (provided by applicant): Mature neutrophils arise from the hematopoietic stem cell via a series of commitment steps. The appearance of the secondary granule proteins (SGP) lactoferrin (LF), transcobalamin I (TCI), neutrophil collagenase (NC) and neutrophil gelatinase (NG) marks the commitment to terminal neutrophil differentiation. C/EBPepsilon (C/EBPe) plays a critical role in the coordinate upregulation of SGP genes. Disruption of the C/EBPe gene in mice leads to morphologic and functional defects in neutrophil maturation with a defective transition from the promyelocyte to the myelocyte stage. The neutrophils have bilobed nuclei, abnormal respiratory burst activity, and impaired chemotaxis and bactericidal activity. They lack specific granules and fail to express mRNAs encoding for secondary and tertiary granule content proteins. The mice die within 3-5 months of infection or from complications of "myeloproliferation". Phenotypic and functional defects of the C/EBPe -/- mice closely parallel those in patients with secondary granule deficiency. We have made 2 different cell lines from the bone marrow of the C/EBPe -/- mice and corresponding wildtype littermates that mimic the morphologic and functional defects in the knockout mice. Using these cell lines and primary marrow cells, we propose to further characterize the transcriptional regulation of terminal neutrophil differentiation and specifically the role of C/EBPe in the neutrophil maturation program. Our specific aims are: 1) To complete the characterization of the newly generated cell lines and establish them as a faithful model of the C/EBPE -/- phenotye: 2) To identify downstream targets of C/EBPepsilon through microarray analysis of the cell lines and primary C/EBPe +/+ and -/- bone marrow; and 3) To rescue the C/EBPepsilon -/- phenotype by retroviral transduction with candidate genes identified in specif aim 2 . Genes will be transferred first into the C/EBPe -/- cell line to determine reversal of phenotype. Verified important targets will be transduced into C/EBPe -/- marrow progenitors and transplanted back into C/EBPe -/- mice to assess reversal of phenotype

描述（由申请人提供）： 成熟的中性粒细胞是由造血干细胞通过一系列承诺步骤引起的。二次颗粒蛋白（SGP）乳铁蛋白（LF），经胆质I（TCI），中性粒细胞胶原酶（NC）和中性粒细胞明胶酶（NG）的出现标志着对末端嗜中性粒细胞分化的承诺。 C/EBPEPSILON（C/EBPE）在协调SGP基因上调中起关键作用。小鼠中C/EBPE基因的破坏会导致中性粒细胞成熟的形态和功能缺陷，从叶虫细胞到骨髓细胞阶段有缺陷的过渡。中性粒细胞具有双核核，异常的呼吸爆发活性以及趋化性和杀菌活性受损。他们缺乏特定的颗粒，无法表达编码二级和第三纪颗粒含量蛋白的mRNA。小鼠在感染后的3-5个月内死亡或“骨髓增生”并发症。 C/EBPE - / - 小鼠的表型和功能缺陷与继发性颗粒缺乏患者的患者紧密平行。我们已经从C/EBPE - / - 小鼠的骨髓和相应的野生型窝窝中产生了2种不同的细胞系，它们模仿了基因敲除小鼠中形态和功能缺陷。使用这些细胞系和原代骨髓细胞，我们建议进一步表征末端嗜中性粒细胞分化的转录调节，特别是C/EBPE在中性粒细胞成熟程序中的作用。我们的具体目的是：1）完成新生成的细胞系的表征，并将其确定为C/EBPE - / - 表格的忠实模型：2）通过对细胞系的微阵列分析和主要C/EBPE +/eBPE +/ +/ +/ +/ +和 - bone -bone Bone -Bone Bone Marrow来识别C/EBPEPSILON的下​​游目标； 3）用逆转录病毒转导，用特异性AIM 2中鉴定的候选基因来挽救C/ebpepsilon - / - 表型。基因将首先转移到C/EBPE - / - 细胞系中，以确定表型的逆转。经过验证的重要目标将被转导为C/EBPE - / - 骨髓祖细胞，并将其移回C/EBPE - / - 小鼠以评估表型的反转