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：
9278153
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/9278153
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 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 Hematologic Neoplasms Hematological Disease Hematopoiesis Hematopoietic Hematopoietic stem cells Impairment In Situ Hybridization Individual Investigation 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 Signaling Molecule Signaling Protein Specificity Stem cells Technology TimeLine Validation WNT Signaling Pathway Zebrafish base bone morphogenetic protein receptors cell type chromatin immunoprecipitation combinatorial database query embryonic stem cell experimental study in vivo insight knock-down member notch protein novel overexpression precursor cell protein activation public health relevance small molecule inhibitor 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）：造血研究对于理解血液疾病的病因至关重要，多年来，多年来一直是回答有关茎和祖细胞遗传调节的基本问题的范式，以及谱系命运。该提案的目的是继续和扩展我们对骨形态发生蛋白（BMP）信号通路的造血分子控制的当前研究，这是胚胎模式的重要调节剂。 BMP是信号分子的大型超家族的成员，并通过激活下游SMAD转录因子的子集发挥其许多生物学作用。我们最近使用小鼠胚胎干（ES）细胞模型的研究表明，SMAD1在造血潜力上发挥了暂时的调节作用，在抑制造血性膨胀之前，先于先前抑制前体细胞之前刺激造血作用。我计划以这些研究为基础，以使用小鼠ES细胞和斑马鱼（Danio Rerio）模型系统来识别和检查关键调节基因网络。为此，我将追求三个单独的目标，这将融合他们对BMP途径的分析以及对各种已建立和新兴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研究BMP效应子SMAD1和SMAD5对造血谱系承诺的个体和组合贡献，并使用小鼠ES细胞中的条件基因敲低策略进行扩展。在第二个目标中，我将使用斑马鱼模型来探测先前鉴定的SMAD1和SMAD5靶标的造血功能，并着眼于定义它们在造血过渡中的作用，这些造血过渡是在发育中发育的胚胎中迁移的活性造血的位置。我将通过鼠ES细胞研究对这些发现进行补充，以通过染色质免疫沉淀来验证直接的SMAD结合靶标。最后，在第三个目的中，我将在ES细胞中使用小抑制剂分子和斑马鱼检查BMP信号在SMAD激活上游的胚胎造血作用的调节作用，并特别注意SMAD-独立过程，以定义BMP介导的特定血统控制的新机制。