Controlling Hematopoietic Lineage Commitment from ESC to Platelets

控制从 ESC 到血小板的造血谱系承诺

基本信息

批准号：
7939700
负责人：
BEVERLY J. TOROK-STORB
金额：
$ 114.82万
依托单位：
FRED HUTCHINSON CANCER RESEARCH CENTER
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2016-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7939700
关键词：
Animals Biological Assay Biology Blood Cells Blood Component Removal Blood Platelets Bone Marrow Transplantation Canis familiaris Cell Lineage Cell Therapy Cell physiology Cells Chemicals Clinical Data Dimerization Educational process of instructing Effectiveness Experimental Models Flow Cytometry Freezing Gene Expression Gene-Modified Generations Genes Genetic Goals Hematopoiesis Hematopoietic Hematopoietic stem cells Hospitals Housing Human In Vitro Intervention Knowledge Libraries Life Linear Accelerator Radiotherapy Systems Liquid substance Magnetic Bead Technology Mammals Megakaryocytes Megakaryocytopoieses Messenger RNA Modeling Molecular Molecular Profiling Mus Nitrogen Operative Surgical Procedures Outcome Pathway interactions Pediatric Hospitals Pharmaceutical Preparations Phase Philadelphia Platelet Count measurement Population Pre-Clinical Model Principal Investigator Production Proteins RNA Interference Reagent Regulator Genes Reporter Research Research Personnel Resources Safety Screening procedure Signal Transduction Signaling Molecule Small Interfering RNA Speed Staging Stem cells Testing Thrombocytopenia Thrombopoiesis Translating Transplantation Treatment Efficacy Virus assay development base cell growth cytopenia design gene therapy genome-wide high throughput screening improved in vivo in vivo Model irradiation man pre-clinical progenitor programs promoter reconstitution response scale up small hairpin RNA small molecule tool vapor vector

项目摘要

DESCRIPTION (provided by applicant): Marrow transplantation have taught us that hematopoiesis is supported by multipotent hematopoietic stem cells that can maintain or reconstitute the various blood cell lineages throughout life. Despite advances in our understanding of lineage commitment, this has not translated into improved approaches for treating lineage specific cytopenias. The goal of our application is to further our understanding of lineage commitment and use this knowledge to develop molecular interventions that will drive hematopoiesis toward desired lineages both in vitro and in vivo. The application is specifically focused on megakaryopoiesis, because of the clinical importance of post-transplant thrombocytopenia, and to maximize synergy with our UOl partners at Childrens Hospital of Philadelphia. Towards this goal we propose a multifaceted program with 4 Aims, with each aim involving two or more investigators from the FHCRC/UW Consortium. In Aim 1, Drs Fero, Paddington and Torok-Storb will generate molecular profiles of functionally defined progenitors in the stem cell to platelet pathway from mouse, dog, and man. These three experimental models have complementary strengths that allow for a comprehensive approach including a robust preclinical in vivo model that can predict clinical outcomes. In Aim 2 these same investigators will gene modify the defined progenitor cells to express lineage-stage-specific reporters for use in a high content, high through put siRNA screening assays to identify changes in the microenvironment that will control progenitor fate. A third Aim will test the proliferation and differentation potential of distinct progenitor subsets in response to conditionally activated signaling molecules. For this purpose Drs Blau and Emery will use signaling molecule derivatives that can be activated in response to small molecule drugs called chemical inducers of dimerization (CIDs). In Aim 4, Drs Kiem and Blau will exploit the canine model of thrombocytopenia to test gene products identified in Aim 2, and cell products expanded in Aim 3 to improve the platelet count in vivo. These in vivo studies in the dog model will establish the safety and efficacy of these therapies in a highly relevant preclinical model

描述（由申请人提供）：骨髓移植告诉我们，造血由多能造血干细胞支持，这些干细胞可以在一生中维持或重建各种血细胞谱系。尽管我们对谱系承诺的理解取得了进步，但这并未转化为治疗谱系特定细胞质的改进方法。我们应用的目的是进一步了解谱系承诺，并利用这些知识来开发分子干预措施，这些干预措施将促使造血在体外和体内朝着所需的谱系。由于移植后血小板减少症的临床重要性，并最大程度地与我们在费城儿童医院的UOL合作伙伴最大程度地相关，因此该应用程序专门集中在巨型近代。为了实现这一目标，我们提出了一个具有4个目标的多面计划，每个目标都涉及来自FHCRC/UW财团的两个或更多研究者。在AIM 1中，Fero博士，Paddington和Torok-Storb将产生干细胞中功能定义的祖细胞的分子谱，从小鼠，狗和人产生血小板途径。这三个实验模型具有互补的强度，可以采用全面的方法，包括可以预测临床结果的强大临床前体内模型。在AIM 2中，这些研究者将基因修改定义的祖细胞，以表达谱系阶段特异性的记者，用于高含量，通过放置siRNA筛选测定，以确定将控制祖细胞命运的微环境的变化。第三个目标将在有条件地激活的信号分子中测试不同祖细胞集的增殖和区分潜力。为此，Drs Blau和Emery将使用信号分子衍生物，这些衍生物可以响应于称为化学诱导剂的二聚化化学诱导剂（CID）而被激活。在AIM 4中，Kiem和Blau博士将利用血小板减少症的犬类模型来测试AIM 2中鉴定的基因产物，而AIM 3中扩展的细胞产物以改善体内的血小板计数。这些在狗模型中的体内研究将在高度相关的临床前模型中确定这些疗法的安全性和功效