Mechanisms of a GATA Factor-Dependent Hematopoietic Signaling Pathway

GATA 因子依赖性造血信号通路的机制

• 批准号: 9295276
• 负责人: Kyle J Hewitt
• 金额: $ 10.01万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9295276
• 关键词: Actin-Binding Protein; Attenuated; Award; Biology; Bone Marrow; Bone Marrow Transplantation; Career Mobility; Cells; Cellularity; ChIP-seq; Core Facility; Correlative Study; Data; Data Set; Defect; Development; Elements; Embryo; Enhancers; Ensure; Environment; Erythroid; Erythropoiesis; Exhibits; Faculty; Fetal Liver; Foundations; G Protein-Coupled Receptor Signaling; Gene Targeting; Genes; Genetic; Goals; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; Hepatocyte; Institution; K-Series Research Career Programs; Knockout Mice; Laboratories; Laboratory Research; Libraries; Mediating; Mentors; Methods; Modeling; Molecular; Mus; Neurabin; Pathology; Physiology; Play; Positioning Attribute; Process; Property; Proto-Oncogene Protein c-kit; Proto-Oncogene Proteins c-akt; Receptor Protein-Tyrosine Kinases; Regulation; Research; Role; SAM Domain; Scientist; Signal Pathway; Signal Transduction; Site; Solid; Spleen; Stem Cell Factor; Stem cells; Stress; Techniques; Testing; Therapeutic; Training; Training Programs; United States National Institutes of Health; Up-Regulation; Work; Writing; career; career development; cohort; design; genome-wide; in vivo; innovation; knock-down; meetings; mouse model; novel; novel strategies; progenitor; programs; regenerative; research and development; self-renewal; skills; small hairpin RNA; success; transcriptome sequencing

Abstract Deleting an E-box-GATA intronic cis-element (+9.5) reduces GATA-2 expression and inactivates the hematopoietic stem cell (HSC) generator in the mouse embryo by disrupting GATA-2-dependent genetic networks. Many of the essential constituents of this genetic network have not been defined. To discover components of this network, the +9.5 sequence/molecular properties were compared to thousands of +9.5-like cis-elements genome-wide. This innovative training method yielded a genome-wide library of +9.5-like sites, containing GATA-2 target genes that may inform unique modes of controlling hematopoietic stem and progenitor cells (HSPCs). I discovered that GATA-2 regulates expression of the unstudied Sterile Alpha Motif Domain 14 (Samd14) gene, and Samd14 increased fetal liver HSPC levels and promoted SCF/c-Kit signaling. Initial data demonstrates that the +9.5-like Samd14-Enhancer mediates Samd14 expression, is required for stress erythropoiesis, and confers stress-dependent Samd14 upregulation in the spleen. These aims will rigorously establish the mechanism by which Samd14 promotes SCF/c-Kit signaling (Aim 1), the function of the GATA-2-regulated Samd14-Enh in an innovative mouse model (Aim 2), and define a cohort of stress- dependent genes and their interconnectivity (Aim 3). As a K award recipient, I will follow a detailed training plan with specific research and career development milestones to be achieved over the course of the award. The Aims are designed to expand and distinguish my research expertise from my mentor, and we have agreed that I can use the Samd14 work as a foundation for developing my independent research program. The career development activities outlined in the proposal will provide training in writing/presentation skills, laboratory management, and grantsmanship. I have organized a mentoring committee comprised of well-established and successful scientists, with research programs relevant to my work, who are committed to my success and will play an active role in facilitating my career transition. A key element of this plan entails direct training in bone marrow transplantation techniques to study stress hematopoiesis. UW-Madison is a top-tier research institution with outstanding NIH-sponsored training programs, core facilities, and research laboratories, which makes it an ideal training environment for me to further develop as an independent scientist. I have strong support from UW-Madison, the Cell and Regenerative Biology Department and Dr. Bresnick to pursue these career goals and compete for a faculty position here or at another research-intensive academic institution. The research Aims, career development training plan, meetings with mentoring committee and collaborators, educational opportunities, and training environment will ensure the development of a solid foundation to launch a productive academic career.

摘要 删除E-box-加塔内含子顺式元件（+9.5）减少了加塔-2表达，并使GATA失活。 通过破坏加塔-2依赖的遗传基因， 网络.这个遗传网络的许多基本组成部分还没有被定义。发现 作为该网络的组成部分，将+9.5序列/分子特性与数千个类似+9.5序列/分子特性进行了比较。 全基因组的顺式元件。这种创新的训练方法产生了一个全基因组的+9.5样位点库， 含有加塔-2靶基因，该靶基因可提供控制造血干细胞的独特模式， 祖细胞（HSPCs）。我发现加塔-2调节未研究的不育α基序的表达 结构域14（Samd14）基因和Samd14增加胎肝HSPC水平并促进SCF/c-Kit信号传导。 初始数据表明，+9.5样Samd14增强子介导Samd14表达，是 应激红细胞生成，并赋予脾中应激依赖性Samd 14上调。这些目标将 严格建立Samd 14促进SCF/c-Kit信号传导的机制（目标1）， 加塔-2调节的Samd 14-Enh在一个创新的小鼠模型（目的2），并定义了一个队列的压力- 依赖基因及其相互连接性（目标3）。 作为K奖获得者，我将遵循一个详细的培训计划与具体的研究和职业发展 在授标过程中要达到的里程碑。目标旨在扩大和区分我的 我的导师的研究专业知识，我们已经同意，我可以使用Samd 14的工作作为基础， 发展我的独立研究计划。建议中概述的职业发展活动将 提供写作/演讲技巧、实验室管理和培训。我组织了一个 指导委员会由知名和成功的科学家组成，具有相关的研究项目 他们致力于我的成功，并将在促进我的职业过渡方面发挥积极作用。一 该计划的关键要素是骨髓移植技术的直接培训，以研究压力 造血威斯康星大学麦迪逊分校是一个顶级的研究机构，具有杰出的NIH赞助的培训 课程，核心设施和研究实验室，这使其成为我理想的培训环境， 进一步发展成为独立的科学家。我有来自威斯康星大学麦迪逊分校，细胞和 再生生物学系和博士bresnick追求这些职业目标，并为教师竞争 在这里或在另一个研究密集型学术机构的位置。研究目的，职业发展 培训计划、与指导委员会和合作者的会议、教育机会和培训 环境将确保发展一个坚实的基础，开展富有成效的学术生涯。