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Molecular Circuits in the Hematopoietic Stem Cell Niche

造血干细胞生态位中的分子回路

基本信息

批准号：
10231033
负责人：
JAMES J COLLINS
金额：
$ 166.57万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-15 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10231033
关键词：
ATAC-seq Adipocytes Adult Algorithms Anatomy Aorta Atlases Blood Bone Marrow Bone Marrow Transplantation Cataloging Catalogs Cell Adhesion Molecules Cell Communication Cell Count Cell Culture Techniques Cell Differentiation process Cell Line Cell Maintenance Cell surface Cells Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Coculture Techniques Code Collaborations Communities Computational algorithm Computer software DNA Binding DNA cassette Data Data Set Derivation procedure Development Ecosystem Elements Embryo Embryonic Development Endothelial Cells Endothelium Engineering Equilibrium Fetal Liver Fishes Gene Expression Gene Expression Profile Gene Transfer Generations Genes Genetic Transcription Glean Goals Gonadal structure Hematology Hematopoiesis Hematopoietic Hematopoietic stem cells Homing Human In Situ Hybridization In Vitro Investigational Therapies Life Location Mammals Mesenchymal Mesonephric structure Mining Modeling Molecular Mouse Strains Mus Neural Crest Placenta Pluripotent Stem Cells Population Pregnancy Recreation Regulator Genes Reporter Reporting Research Research Personnel Resolution Resources Shapes Signal Transduction Site Somites Source Specific qualifier value System Techniques Testing Therapeutic Transplantation Umbilical Cord Blood Work Zebrafish cell type computational pipelines critical period data resource fetal gene discovery genetic manipulation hematopoietic differentiation hematopoietic stem cell differentiation hematopoietic stem cell emergence hematopoietic stem cell expansion hematopoietic stem cell niche hemogenic endothelium in vivo innovation insight intercellular communication knock-down macrophage novel open source programs recruit self-renewal single-cell RNA sequencing stem cell migration stem cells synthetic biology synthetic construct transcriptome sequencing user-friendly web site

项目摘要

In mammals, hematopoietic stem cells (HSCs) first arise from a specialized hemogenic endothelium that lines the developing embryonic aorta, migrate to and expand in the fetal liver, and ultimately colonize the bone marrow, which supports hematopoiesis throughout adult life. These distinct anatomic locations harbor specialized microenvironments that support the developmental maturation, expansion, and ultimately the balance of self-renewal and differentiation of HSCs. The transcriptional programs that promote formation and differentiation of hematopoietic stem and progenitor cells (HSPCs) have been widely interrogated, but much remains to be learned about the supportive niche cells of the hematopoietic microenvironment and the mechanisms of cell-cell interaction that specify HSC emergence during development, HSC migration, lodging, and expansion in fetal niches, and the ultimate quiescence, self-renewal, and differentiation in the bone marrow. In our preliminary data, we have gathered evidence for number of cell types, including endothelial cells, mesenchymal cells, macrophages, neural crest derivatives, and somites as components of the hematopoietic niche. We will gather comprehensive “omics” data to catalogue the gene expression programs within the distinct hematopoietic niche cells that occur during development in the aorta-gonad-mesonephros (AGM), fetal liver, bone marrow, and placenta (aim 1). Our approach begins with tomo-seq, which enables us to discover gene expression patterns unique to cell populations like endothelium that have region-specific specialization. We will validate cell-specific expression in FACS purified cells by single cell RNA-seq and in situ hybridization, and will document functionality using morpholino and CRISPR knock-down in the experimentally tractable zebrafish model. We then use ATAC-seq to define functional open chromatin around these genes, and motif-finding software to identify DNA-binding regulatory factors that are candidate drivers of hematopoietic cell fate. We will employ a computational pipeline and develop novel algorithms to analyze these data (aim 2). Hypotheses emerging from aims 1 and 2 will be tested by constructing novel reporter strains of zebrafish and mice, as well as engineered pluripotent stem cells carrying synthetic reporters and drivers (aim 3). Our goal is to define the molecular circuitry that specifies niche cells during the critical periods of HSC emergence and expansion, and to probe cross-talk between niche elements and HSPCs. We hope to glean unique insights into the molecular mechanisms that drive hematopoietic formation and maturation during embryonic development, and to enhance our understanding of HSC maintenance, quiescence, self-renewal and differentiation.

在哺乳动物中，造血干细胞（hsc）首先产生于一种特殊的造血内皮细胞