Specification of blood lineages in a vertebrate embryo

脊椎动物胚胎血统的规范

• 批准号: 7086219
• 负责人: ROBERT K HO
• 金额: $ 27.11万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7086219
• 关键词: alternatives to animals in research; biological signal transduction; cell differentiation; cell growth regulation; cell proliferation; cell transplantation; developmental genetics; embryogenesis; embryonic stem cell; erythrocytes; gene environment interaction; gene expression; genetic models; genetic regulation; genetically modified animals; hematopoietic stem cells; mesoderm; mutant; myelocyte; transcription factor; vertebrate embryology; zebrafish; alternatives to animals in research; biological signal transduction; cell differentiation; cell growth regulation; cell proliferation; cell transplantation; developmental genetics; embryogenesis; embryonic stem cell; erythrocytes; gene environment interaction; gene expression; genetic models; genetic regulation; genetically modified animals; hematopoietic stem cells; mesoderm; mutant; myelocyte; transcription factor; vertebrate embryology; zebrafish

DESCRIPTION (provided by applicant): The overall goals of this proposal are to characterize the lineal origins of blood cells and to determine the effects of cell interactions upon fate decisions made by vertebrate blood cell precursors. Our identification of separate sites of production of macrophages and red blood cells in the zebrafish will allow for a dissection of the different environmental and genetic influences that cause precursors to adopt myeloid and erythroid fates. We have identified a key interaction with the paraxial mesoderm that is required for proper RBC formation and will further study the implications of this interaction for future diagnostic and therapeutical implications related to leukemias, myelomas, lymphomas, myelodysplastic syndromes and anemias. An understanding of the influences that affect cell fate choices will potentially be important in the screening, culturing and in vitro differentiation of stem cells for such procedures as bone marrow transplants and the treatment of genetic disorders such as thalassemia and sickle cell anemia. Our general strategy is to use the genetic and cellular advantages of the model system zebrafish to understand blood cell differentiation. The zebrafish has become a viable and important model system for blood cell differentiation in amniotes, including humans. There are currently over 50 described zebrafish mutations in 26 complementation groups that exhibit blood defects. Characterization and cloning of such hematopoietic mutants has made substantial contributions to human disease research, for instance the zebrafish mutant, sauternes, currently serves as the only model of human congenital sideroblastic anemia. Transgenic lines of zebrafish serve as model systems for cancer studies, for instance, lines of zebrafish expressing c-myc exhibit symptoms of T-cell acute leukemia. The zebrafish mutant spadetail fails to form early Red Blood Cell lineages but retains the ability to form early macrophage lineages. We propose to study the role of both cell-autonomous and environmental factors that are required to form the erythroid and myeloid cell fate by comparing the process of blood formation between wild-type and spadetail mutant zebrafish. Specifically, we will perform fate map studies to which the zebrafish is particularly amenable, for the purpose of directly determining the origins of myeloid and red blood cell precursors. We will determine when the spadetail gene function and environmental signals are required to cooperate for proper red blood cell production by transplanting wildtype cells into various positions within the spadetail mutant to rescue the loss of red blood cell phenotype. We will also use cell transplantations to determine if an interaction with paraxial mesoderm can respecify myeloid precursors to adopt a red blood cell fate.

描述（由申请人提供）：该提案的总体目标是表征血细胞的线性起源，并确定细胞相互作用对脊椎动物血细胞前体做出的命运决定的影响。我们鉴定斑马鱼中巨噬细胞和红细胞生产的单独生产部位将允许解剖不同的环境和遗传影响，从而导致前体采用髓样和红细胞命运。我们已经确定了适当的RBC形成所必需的与近期中胚层的关键相互作用，并将进一步研究这种相互作用对与白血病，脊髓瘤，淋巴瘤，骨髓异常成年综合征和贫血相关的将来诊断和治疗意义的含义。了解影响细胞命运选择的影响的理解对于骨髓移植和诸如thalassalasealia和镰状细胞贫血等遗传疾病的治疗等过程的筛查，培养和体外分化可能很重要。 我们的一般策略是使用模型系统斑马鱼的遗传和细胞优势来了解血细胞的分化。斑马鱼已成为包括人类在内的羊膜细胞分化的可行且重要的模型系统。目前有50多个描述的斑马鱼突变在26个互补组中表现出血液缺陷。这种造血突变体的表征和克隆对人类疾病研究做出了重大贡献，例如斑马鱼突变体Sauternes，目前是人类先天性SiderOlbastic贫血的唯一模型。斑马鱼的转基因线充当癌症研究的模型系统，例如，表达C-Myc的斑马鱼线表现出T细胞急性白血病的症状。斑马鱼突变尾尾无法形成早期的红细胞谱系，但保留了形成早期巨噬细胞谱系的能力。我们建议研究通过比较野生型和小尾突变突变体斑马鱼之间的血液形成过程来研究形成红细胞和髓样细胞命运所需的细胞自主和环境因素的作用。 具体而言，我们将进行斑马鱼特别适合的命运图研究，目的是直接确定髓样和红细胞前体的起源。我们将通过将野生型细胞移植到Spadetail突变体中的各种位置，以挽救红细胞表型的丧失，从而确定何时需要使用Spadetail基因功能和环境信号来合作以进行适当的红细胞产生。我们还将使用细胞移植来确定与副胚层的相互作用是否可以重新定位髓样前体以采用红细胞命运。