Role of slug in hematopoietic stem cell regeneration

蛞蝓在造血干细胞再生中的作用

基本信息

批准号：
8338402
负责人：
WEN-SHU WU
金额：
$ 35.24万
依托单位：
CHILDREN'S HOSPITAL & RES CTR AT OAKLAND
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-23 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8338402
关键词：
Adult Aplastic Anemia Area Attenuated Binding Binding Sites Biological Assay Blood Cells Blood Vessels Bone Marrow Bone Marrow Transplantation Cell Culture Techniques Cell Cycle Cell division Cells Cleaved cell Clinical Treatment Consensus Cyclin D1 Data Disease Down-Regulation Dyes Feedback Fluorouracil Gene Expression Genes Genetic Hematopoiesis Hematopoietic Hematopoietic Stem Cell Transplantation Hematopoietic System Hematopoietic stem cells Human In Vitro Injection of therapeutic agent Injury Intervention Luciferases MMP9 gene Mediating Membrane Messenger RNA Methods Modeling Molecular Mus Natural regeneration Patients Production Proto-Oncogene Protein c-kit Publishing Recovery Regulation Relative (related person)Reporter Reporter Genes Research Residual state Role Sickle Cell Anemia Signal Pathway Signal Transduction Stem Cell Factor Stress Testing Therapeutic Time Transcription Repressor/Corepressor Transgenes Transplantation Umbilical Cord Blood Work Zinc Fingers base blood resource c-myc Genes cell type improved in vivo inhibitor/antagonist innovation leukemia mouse model notch protein novel novel strategies peripheral blood premature prevent promoter response self renewing cell self-renewal slug small hairpin RNA stem cell division success

项目摘要

DESCRIPTION (provided by applicant): Hematopoietic stem cells (HSCs) are rare self-renewing cells in the hematopoietic compartment that fuel the production of millions of differentiated blood cells of various types. HSC transplantation has become a common clinical treatment for patients suffering from leukemia, aplastic anemia, sickle cell disease, and other disorders of hematopoietic systems. Because transplantation success depends heavily on the number of available HSCs in peripheral and cord blood resources, expansion of HSCs both in vivo and in vitro has assumed a high priority. Unfortunately, progress in this research area has been impeded by the limited understanding of the molecular mechanisms governing the decision of HSCs to self-renew or differentiate. In the proposed project, we aim to define the role of Slug, an evolutionarily conserved zinc-finger transcriptional repressor, in the regulation of HSC self-renewal. Our most recent work shows that Slug is dispensable for normal hematopoiesis, but is critical for suppression of blood cell repopulation after bone marrow transplantation or treatment with 5-fluorouracil (5- FU). Additional studies suggest that Slug is a potent negative regulator of c-kit and other cell cycle regulators (including cyclin D1, c-Myc, and Id3), while it is positively regulated by the SCF/c-Kit signaling in HSCs. These findings led logically to the central hypothesis of the current application: that Slug functions in a novel negative- feedback loop in SCF/c-kit signaling pathway, and that its deletion will promote symmetric division of HSCs during repopulation of the hematopoietic systems. These predictions will be tested by examining the effects of Slug deficiency on the integrity of HSCs following myelotoxic injury (Aim 1); by determining if Slug regulates symmetric cell division of HSCs during hematopoietic regeneration (Aim 2); and by elucidating Slug's role in the SCF/c-kit signaling pathway (Aim 3). Successful completion of these aims will be facilitated by innovative methods and strategies that were developed specifically for this project, including single cell division assays based on PKH2-26 dye and Notch-GFP reporters to discriminate between symmetric and asymmetric division, and a new HSC self-renewal competitive assay. The information we expect to generate will have the potential to improve HSC expansion both in vitro and in vivo.

描述（由申请人提供）：造血干细胞（HSC）是造血区室中罕见的自我更新细胞，为数百万种不同类型的分化血细胞的产生增添了生产。 HSC移植已成为患有白血病，性障碍性贫血，镰状细胞疾病和其他造血系统疾病的患者的常见临床治疗方法。由于移植成功在很大程度上取决于外围和脐带血资源中可用的HSC的数量，因此体内和体外的HSC的扩展具有很高的优先级。不幸的是，该研究领域的进步受到了对HSC对自我更新或分化的决定的分子机制有限的理解的阻碍。在拟议的项目中，我们旨在定义Slug（进化保守的锌指转录阻遏物）在HSC自我更新调节中的作用。我们最近的工作表明，SLUG对于正常的造血是可分配的，但对于用5-氟尿嘧啶（5- FU）治疗后抑制血细胞再生物至关重要。其他研究表明，SLUG是C-KIT和其他细胞周期调节剂（包括Cyclin D1，C-MYC和ID3）的有效负调节剂，而HSC中的SCF/C-KIT信号则对其进行了积极调节。这些发现从逻辑上导致了当前应用的中心假设：SLUG在SCF/C-KIT信号通路中的新型负反馈循环中起作用，并且其缺失将促进造血系统重生过程中HSC的对称分裂。这些预测将通过检查骨髓毒性损伤后的SLUG缺乏对HSC的完整性的影响来测试这些预测（AIM 1）；通过确定SLUG在造血再生过程中是否调节HSC的对称细胞分裂（AIM 2）；并通过阐明Slug在SCF/C-KIT信号通路中的作用（AIM 3）。这些目标的成功完成将通过专门为该项目开发的创新方法和策略来促进，包括基于PKH2-26染料和Notch-GFP记者的单细胞分裂测定，以区分对称和非对称分区，以及一种新的HSC自我更新竞争力的竞争力。我们期望产生的信息将有可能在体外和体内改善HSC扩展。