The zinc finger transcription factor ZBP89 in blood & vascular development

血液中的锌指转录因子ZBP89

• 批准号: 8287061
• 负责人: M. AMIN ARNAOUT
• 金额: $ 37.3万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8287061
• 关键词: Adult; Alleles; Anemia; B-Lymphocytes; Biochemical; Biological Assay; Biotinylation; Blast Cell; Blood; Blood Cells; Blood Vessels; Bone Marrow; Bone Marrow Cells; Bone Marrow Transplantation; C2H2 Zinc Finger; CCAAT-Enhancer-Binding Protein-alpha; CCAAT-Enhancer-Binding Proteins; Candidate Disease Gene; Cell Culture Techniques; Cell Line; Cell Lineage; Cells; Chronic; Commit; Complementary DNA; Data; Defect; Development; EMSA; ES Cell Line; Ectopic Expression; Embryo; Endothelial Cells; Erythrocytes; Erythroid; Exhibits; Failure; Fishes; GATA1 gene; Gene Expression; Generations; Genes; Genetic; Genomics; Health; Helix-Turn-Helix Motifs; Hematopoiesis; Hematopoietic; Hematopoietic System; Hematopoietic stem cells; Human; Hybrids; ITGAM gene; In Vitro; Inflammation; Injection of therapeutic agent; Integrins; KDR gene; Knock-out; Knockout Mice; L-Plastin; Leukocytosis; Link; Lymphoid; MYB gene; Mediating; Mesoderm; Methods; Modeling; Molecular; Mus; Mutagenesis; Myelogenous; Myeloid Cells; Myocardial Infarction; Nature; Orthologous Gene; PECAM1 gene; PTPRC gene; Perinatal mortality demographics; Phenotype; Play; Population; Process; Program Development; Proteins; Proteomics; Proto-Oncogene Proteins c-myb; RNA Caps; Radiation; Reporter; Role; Series; Specific qualifier value; Staging; Stem cells; TAL1 gene; TIE gene; Transfusion; Transgenic Organisms; Transplantation; Undifferentiated; Yeasts; Zebrafish; Zinc Fingers; angiogenesis; cadherin 5; cardiac repair; cell type; chemotherapy; developmental plasticity; embryonic stem cell; fetal; human GATA1 protein; in vivo; macrophage; monocyte; mutant; neutrophil; novel strategies; overexpression; progenitor; programs; promoter; protein complex; reconstitution; self-renewal; small hairpin RNA; transcription factor; tumor

DESCRIPTION (provided by applicant): The hematopoietic system originates from a small population of self-renewing hematopoietic stem cells (HSCs) that in turn derive from hemangioblasts, common precursors of blood and blood vessels. Extensive studies have identified key transcription factors, such as the basic helix loop helix (bLH) SCL/TAL1 and GATA factors, which play critical roles in the successive steps of differentiation of HSCs into the erythroid, myeloid and lymphoid lineages. Despite these advances, the nature of the factors that commit the hemangioblast to blood and endothelial cells are obscure and the transcriptional networks operative in HSC self-renewal and lineage commitment remain incomplete. In preliminary studies, we have identified the Kruppel-like zinc finger transcription factor ZBP-89 as a master regulator of early development of FLK1+ mesoderm into blood and blood vessels. Forced expression of ZBP-89 increased expression of primitive and definitive hematopoiesis in zebrafish and in mouse embryonic stem cells (ESCs)/embryonic body (EB) cultures, but reduced axial and intersomitic blood vessel formation in zebrafish embryos and the endothelial replating potential and sprouting angiogenesis in mouse EB cultures. Knockdown of ZBP-89 in zebrafish or in mouse ESCs resulted in a dramatic reduction in primitive and definitive hematopoietic markers (e.g. SCL and GATA factors), but an increase in endothelial lineage markers, distinguishing the mechanism of action of ZBP-89 from all other known transcription factors regulating hematovascular development. Further, we find that mouse pubs homozygous for a hypomorphic ZBP-89 mutant allele die perinatally and exhibit a marked reduction in circulating red blood cells, but an increase in mature myeloid cells, reflecting an additional critical role for ZBP-89 in fetal hematopoiesis, perhaps at the level of the bipotential Common Myeloid Progenitor (CMP) stem cell, which gives rise to both the erythroid and myeloid lineages. The effects of ZBP-89 modulation on adult hematopoiesis and the underlying transcriptional networks involved in its action at the level of hemangioblasts and CMPs are unexplored. In this application, we propose to assess the consequences of loss of ZBP-89 on fetal and adult hematopoiesis and vascular development in vivo (Aim 1), determine the effects of its ectopic expression on blood and vessel lineage development in adult mice and zebrafish (Aim 2), and elucidate the mechanism(s) underlying ZBP-89- mediated hematopoietic lineage commitment. (Aim 3). Genetic, biochemical, proteomics, genomics, stem cell cultures and bone marrow transplantation in mouse and zebrafish will be utilized. PUBLIC HEALTH RELEVANCE: Formation of all blood cells is closely linked developmentally to formation of blood vessels and the two processes are regulated in large part by transcription factors, proteins that control the decision of a common undifferentiated stem cell precursor to become a blood cell or a vascular cell. The instructional network that underlies this cell fate decision remains ill defined, despite its potential impact on development of new therapies. For example, blood stem cells are being used to reconstitute bone marrow damaged by radiation or chemotherapy, but they are very few in numbers, a major challenge in cellular therapy. Increasing their number by enhancing their development from their stem cell precursors will serve a critical need. Methods for enhancing or suppressing new blood vessel formation (angiogenesis) may also be useful respectively, in cardiac repair following a heart attack or in reducing harmful angiogenesis (induced by tumors or chronic inflammation). We have discovered a transcription factor that lies at the cross road of blood and blood vessel development. We are proposing a series of studies to pinpoint the role of this factor, and define the developmental networks it coordinates.

描述（由申请人提供）：造血系统起源于少数自我更新的造血干细胞（HSC），这些造血细胞（HSC）反过来源自血管细胞，血管细胞，是血管和血管的常见前体。广泛的研究已经确定了关键的转录因子，例如基本的螺旋环螺旋（BLH）SCL/TAL1和GATA因子，这些因子在HSC的连续分析步骤中起着至关重要的作用，将HSC分化为红系，髓样和淋巴样谱系。尽管有这些进展，但将血管细胞归于血管细胞和内皮细胞的因素的性质是晦涩难懂的，并且在HSC自我更新和谱系承诺中操作的转录网络仍然不完整。在初步研究中，我们已经确定了Kruppel样锌指转录因子ZBP-89是FLK1+中胚层早期发育中的主要调节剂。 ZBP-89的强迫表达增加了斑马鱼和小鼠胚胎干细胞（ESC）/胚胎体（EB）培养物中原始和确定的造血的表达，但减少了Zebrafish Embryos中的轴向和肌间血管形成，并在胚胎中以及内皮化的潜在和新生的EBERINE EBERINE EBERUTINE EBERTINE EBERINE EBERINE EBERENIS IN MELE EBERISERISIS。 Zebrafish或Mouse ESC中ZBP-89的敲低导致原始和确定的造血标志物的急剧减少（例如SCL和GATA因子），但内皮谱系标记的增加，从而区分了ZBP-89的作用机制，从而将ZBP-89的作用机制与其他已知转录因子的作用机制区分开。 Further, we find that mouse pubs homozygous for a hypomorphic ZBP-89 mutant allele die perinatally and exhibit a marked reduction in circulating red blood cells, but an increase in mature myeloid cells, reflecting an additional critical role for ZBP-89 in fetal hematopoiesis, perhaps at the level of the bipotential Common Myeloid Progenitor (CMP) stem cell, which gives rise to both the红细胞和髓样谱系。 ZBP-89调节对成年造血的影响以及在血管细胞和CMP水平上涉及其作用的基本转录网络尚未探索。在此应用中，我们建议评估ZBP-89损失对体内胎儿和成人造血的影响以及体内的血管发育（AIM 1），确定其异位表达对成年小鼠和斑马拉夫（AIM 2）血液和血管发育的影响（AIM 2），并在ZBP-89下进行了固定在ZBP-89-89-89-89-89-89-89-89-89-89-89-89-89- （目标3）。将利用小鼠和斑马鱼中的遗传，生化，蛋白质组学，基因组学，干细胞培养物以及骨髓移植。公共卫生相关性：所有血细胞的形成与血管的形成紧密相关，这两个过程在很大程度上由转录因子进行调节，转录因子控制了常见的未分化的干细胞前体的决策成为血细胞或血管细胞。尽管对新疗法的发展潜在影响，但仍在该细胞命运决定的基础的教学网络仍未定义。例如，血液干细胞用于重建受放射或化学疗法损坏的骨髓，但数量很少，这是细胞疗法的主要挑战。通过从干细胞前体增强其发展来增加数量将满足至关重要的需求。增强或抑制新血管形成（血管生成）的方法也可以分别有用，在心脏病发作后心脏修复或减少有害血管生成（由肿瘤或慢性炎症引起）。我们发现了一个位于血管发育十字路口的转录因子。我们提出了一系列研究，以查明该因素的作用，并定义其协调的发展网络。