Role of C/EBPepsilon in myeloid differentiation

C/EBPepsilon 在骨髓分化中的作用

• 批准号: 7802278
• 负责人: Stephanie Halene
• 金额: $ 13.09万
• 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/7802278
• 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; 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; NOD/SCID mouse; Neutrophil Collagenase; Patients; Phagocytosis; Phenotype; Play; Process; Progranulocytes; Proteins; Reporting; Respiratory Burst; Retroviral Vector; Role; 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

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 defectivetransition 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 defectsof 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/EBPepsilonthrough microarray analysis of the cell lines and primary C/EBPe +/+ and -/- bonemarrow; 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 reversalof phenotype. Verified important targets will be transduced into C/EBPe-/- marrow progenitors and transplanted back into C/EBPe-/- mice to assess reversal of phenotype.

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。这 次生颗粒蛋白（SGP）乳铁蛋白（LF），跨番茄I（TCI），中性粒细胞的外观 胶原酶（NC）和中性粒细胞明胶酶（NG）标志着对末端嗜中性粒细胞的承诺 分化。 C/EBPEPSILON（C/EBPE）在协调SGP基因上调中起关键作用。 小鼠中C/EBPE基因的破坏会导致嗜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的缺陷术。嗜中性粒细胞有双眼 核，异常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，无法表达用于二级和三级颗粒含量蛋白的编码的mRNA。 小鼠在感染后的3-5个月内死亡或“骨髓增生”并发症。表型和 C/EBPE - / - 小鼠的功能缺陷与次生颗粒患者的患者紧密平行 不足。我们从C/EBPE - / - 小鼠的骨髓中制成了2种不同的细胞系， 相应的野生型同窝窝，它们模仿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转录 调节末端中性粒细胞分化，特别是C/EBPE在中性粒细胞中的作用 成熟计划。我们的具体目的是：1）完成新生成的单元格的表征 线条并将它们确立为C/EBPE的忠实模型 - / - 表格：2）识别下游目标 C/eBpepsilOnthough的微阵列分析和主要C/EBPE +/ +和 - / - bonemarrow; 3）用鉴定的候选基因逆转录病毒转导拯救C/ebpepsilon - / - 表型 在特定目标2中。基因将首先转移到C/EBPE - / - 细胞系中，以确定逆转 表型。经过验证的重要目标将被引入C/EBPE - / - 骨髓祖细胞和 移回C/EBPE - / - 小鼠以评估表型的逆转。