Discovery of BMP-dependent mechanisms controlling embryonic hematopoiesis

控制胚胎造血的 BMP 依赖性机制的发现

基本信息

批准号：
8504379
负责人：
Brandoch D Cook
金额：
$ 8.74万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-24 至 2018-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8504379
关键词：
Active Sites Acute Address Attention Binding Biological Biological Markers Biological Models Blood Bone Morphogenetic Proteins Cell Count Cell Lineage Cell model Cells Chronic Commit Complement Databases Defect Development Developmental Process Disease E-Box Elements ES Cell Line Embryo Engineering Etiology Experimental Models Eye Flow Cytometry Future Gene Expression Profile Gene Targeting Genes Genetic Genomics Germ Layers Goals Hematological Disease Hematopoiesis Hematopoietic In Situ Hybridization Individual Investigation Malignant Neoplasms Mediating Mesoderm Modeling Molecular Mus Myelogenous Nature Notch Signaling Pathway Palliative Care Pathway interactions Pattern Phenotype Phosphotransferases Population Process Protein Inhibition Publishing RNA Interference Receptor Activation Regulation Regulator Genes Reporter Research Role Signal Pathway Signal Transduction Signaling Molecule Specificity Stem cells Technology TimeLine Validation Zebrafish base bone morphogenetic protein receptors cell type chromatin immunoprecipitation combinatorial embryonic stem cell in vivo inhibitor/antagonist insight member notch protein novel overexpression precursor cell progenitor protein activation public health relevance research study small molecule stem transcription factor transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): The study of hematopoiesis is essential for understanding the etiology of blood diseases, and for many years has served as a paradigm for answering fundamental questions concerning genetic regulation of stem and progenitor cells, and lineage fate. The goal of this proposal is to continue and expand our current research on the molecular control of hematopoiesis by the bone morphogenetic protein (BMP) signaling pathway, an important regulator of embryonic patterning. BMPs are members of a large superfamily of signaling molecules, and exert many of their biological effects via activation of a subset of downstream SMAD transcription factors. Our recent studies using a mouse embryonic stem (ES) cell model have shown that Smad1 exerts temporally distinct regulatory roles on hematopoietic potential, stimulating hematopoiesis prior to commitment of precursor cells before later inhibiting hematopoietic expansion. I plan to build on these studies to identify and examine key regulatory gene networks using both mouse ES cell and zebrafish (Danio rerio) model systems. For this purpose, I will pursue three separate aims, which will converge on their analysis of the BMP pathway and the validation of relevant gene targets identified by various established and emerging genomic technologies and databases. Experiments in the first Aim will investigate the individual and combinatorial contributions of the BMP effectors Smad1 and Smad5 to hematopoietic lineage commitment and expansion using conditional gene knockdown strategies in mouse ES cells. In the second Aim, I will use the zebrafish model to probe the hematopoietic functions of previously identified targets of Smad1 and Smad5, with an eye toward defining their roles in hematopoietic transitions that progress as the site of active hematopoiesis migrates within the developing embryo. I will complement these findings with murine ES cell studies to validate direct SMAD binding targets via chromatin immunoprecipitation. Finally, in the third Aim I will use small inhibitor molecules in ES cells and the zebrafish to examine regulatory roles of BMP signaling on embryonic hematopoiesis upstream of SMAD activation, with particular attention to SMAD- independent processes, to define novel mechanisms of BMP-mediated control of specific blood lineages.

描述（由申请人提供）：造血作用的研究对于理解血液疾病的病因学至关重要，并且多年来一直作为回答有关干细胞和祖细胞的遗传调控以及谱系命运的基本问题的范例。该提案的目标是继续并扩展我们目前对骨形态发生蛋白（BMP）信号通路（胚胎模式的重要调节因子）对造血作用的分子控制的研究。 BMP 是信号分子大家族的成员，通过激活下游 SMAD 转录因子的子集来发挥其许多生物效应。我们最近使用小鼠胚胎干 (ES) 细胞模型进行的研究表明，Smad1 对造血潜力发挥暂时不同的调节作用，在前体细胞定型之前刺激造血，然后抑制造血扩张。我计划在这些研究的基础上，使用小鼠 ES 细胞和斑马鱼（Danio rerio）模型系统来识别和检查关键的调控基因网络。为此，我将追求三个不同的目标，这些目标将集中于对 BMP 途径的分析以及对通过各种已建立和新兴的基因组技术和数据库确定的相关基因靶标的验证。第一个目标中的实验将利用小鼠 ES 细胞中的条件基因敲低策略，研究 BMP 效应子 Smad1 和 Smad5 对造血谱系定型和扩增的个体和组合贡献。在第二个目标中，我将使用斑马鱼模型来探测先前确定的 Smad1 和 Smad5 靶标的造血功能，着眼于定义它们在造血转变中的作用，这些转变随着活跃造血作用位点在发育中胚胎内的迁移而进展。我将通过小鼠 ES 细胞研究来补充这些发现，以通过染色质免疫沉淀验证直接 SMAD 结合靶标。最后，在第三个目标中，我将在 ES 细胞中使用小抑制剂分子，斑马鱼研究 BMP 信号传导对 SMAD 激活上游胚胎造血的调节作用，特别关注 SMAD 独立过程，以确定 BMP 介导的特定血统控制的新机制。