NOTCH SIGNALING IN MYELOID DEVELOPMENT

骨髓发育中的 NOTCH 信号传导

• 批准号: 8424128
• 负责人: Jeffery M Klco
• 金额: $ 12.72万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-15 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8424128
• 关键词: American Society of Hematology; Antibodies; Award; Binding; Biological Assay; C57BL/6 Mouse; Cell Lineage; Cells; ChIP-seq; Chimeric Proteins; Data; Development; Disease; Dominant-Negative Mutation; Epigenetic Process; Equilibrium; Exhibits; Fostering; Funding; Gene Targeting; Genes; Genomics; Goals; Grant; Hematopathology; Hematopoietic; Hematopoietic System; In Vitro; Knowledge; Ligands; Maintenance; Malignant - descriptor; Mediating; Mentors; Methodology; Methods; Monitor; Mouse Strains; Mus; Myelogenous; Myeloid Cells; Myelopoiesis; Myeloproliferative disease; Oncogenes; Pathway interactions; Patients; Pattern; Penetrance; Physicians; Population; Property; Proto-Oncogene Protein c-kit; Publications; RARA gene; Reagent; Research; Resources; Role; Sampling; Scientist; Signal Transduction; Site; Stem cells; T-Lymphocyte; Technology; Training; Universities; Washington; Work; Writing; base; body system; cancer genomics; career; career development; chromatin immunoprecipitation; design; fitness; histone modification; in vivo; in vivo Model; leukemia; mouse model; notch protein; novel; novel therapeutic intervention; progenitor; programs; public health relevance; research study; self-renewal; skills; stem; transcription factor; tumor

DESCRIPTION (provided by applicant): This project describes a 5-year career development program designed to foster an academic career in hematologic research and hematopathology. The proposed research program will build on an existing fund of knowledge using the unique resources available at Washington University, which has a proven track record of developing physician scientists. Dr. Timothy Ley, an expert in myeloid development and cancer genomics and a previous recipient of the American Society of Hematology Mentor Award, will serve as the research mentor. The goals of this proposal are to obtain practical and/or didactic training in scientific methodology, technical skills, grant writing, and scientific publication for the purposeof establishing an independent research program focused on hematopoietic development. The proposed research will focus on the Notch pathway to determine the mechanism(s) by which Notch signaling alters myeloid development. Our recent studies have shown that myeloid progenitors expressing the PML-RARA oncogene (using the Ctsg-PML-RARA mouse model) have enhanced self-renewal, which can be abrogated in vitro by Notch inhibition. These observations will be extended in vivo using an established mouse model of Notch inactivation (conditional DN-MAML-GFP) in the following Aims: Specific Aim 1: We will investigate the role of Notch signaling in self-renewal and myeloid development in vivo. The impact of Notch blockade in both normal and Ctsg-PML-RARA-derived hematopoietic stem/progenitor cells (HSPCs) will be assessed by in vivo methods to assess progenitor fitness and signaling, as well as the long-term development of myeloid neoplasms; and Specific Aim 2: We will identify Notch target genes and epigenetic changes in early myeloid progenitors. The GFP moiety on the DN-MAML1-GFP fusion will be targeted for chromatin immunoprecipitation followed by Illumina paired-end sequencing (ChIP-Seq) to determine the genomic targets of Notch activation in c-Kit positive cells (enriched in myeloid stem/progenitors). The studies will be paired with transcriptional profiling and additional ChIP-Seq experiments to monitor histone modifications. These studies will define the mechanisms by which the Notch pathway influences myeloid development and will identify myeloid-specific Notch functions, which may ultimately guide novel therapeutic approaches for myeloid diseases.

描述（由申请人提供）：该项目描述了一项为期5年的职业发展计划，旨在培养血液学研究和血液病理学的学术职业。拟议的研究计划将使用华盛顿大学可用的独特资源建立在现有的知识基金的基础上，该资源具有开发医师科学家的良好记录。髓样发育和癌症基因组学专家，美国血液学辅导学会奖的先前获得者Timothy Ley博士将担任研究导师。该提案的目标是在科学方法论，技术技能，赠款写作和科学出版物中获得实用和/或教学培训，以建立一个专注于造血发展的独立研究计划。拟议的研究将集中于确定Notch信号改变髓样发育的机制的Notch途径。我们最近的研究表明，表达PML-RARA癌基因（使用CTSG-PML-RARA小鼠模型）的髓样祖细胞增强了自我更新，可以通过Notch抑制在体外消除。这些观察结果将使用已建立的Notch失活的小鼠模型（条件DN-MAML-GFP）在以下目的中进行扩展：具体目标1：我们将研究Notch信号在体内自我更新和髓样发育中的作用。 Notch封锁在正常和CTSG-PML RARA衍生的造血茎/祖细胞（HSPC）中的影响将通过体内方法评估评估祖细胞适应和信号传导的影响，以及髓样肿瘤的长期发展；和特定目标2：我们将确定髓样早期祖细胞中的Notch靶基因和表观遗传变化。 DN-MAML1-GFP融合上的GFP部分将用于染色质免疫沉淀，然后是Illumina配对末端测序（CHIP-SEQ），以确定C-KIT阳性细胞中Notch激活的基因组激活的基因组激活（富集在髓样茎/祖细胞中）。研究将与转录分析和其他芯片zeq实验配对，以监测组蛋白的修饰。这些研究将定义Notch途径影响髓样发育的机制，并确定髓样特异性的缺口功能，这最终可能指导针对髓样疾病的新型治疗方法。