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）：该项目描述了一个为期 5 年的职业发展计划，旨在培养血液学研究和血液病理学方面的学术职业。拟议的研究计划将利用华盛顿大学现有的独特资源建立在现有的知识基金的基础上，华盛顿大学在培养医师科学家方面拥有良好的记录。骨髓发育和癌症基因组学专家、美国血液学会导师奖获得者 Timothy Ley 博士将担任研究导师。该提案的目标是获得科学方法、技术技能、资助写作和科学出版方面的实践和/或教学培训，以建立专注于造血发育的独立研究计划。拟议的研究将重点关注 Notch 通路，以确定 Notch 信号传导改变骨髓发育的机制。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）的骨髓祖细胞具有增强的自我更新能力，可以通过 Notch 抑制在体外消除这种自我更新。这些观察结果将在体内使用已建立的Notch失活小鼠模型（条件性DN-MAML-GFP）进行扩展，具体目标如下： 具体目标1：我们将研究Notch信号传导在体内自我更新和骨髓发育中的作用。将通过体内方法评估Notch阻断对正常和Ctsg-PML-RARA衍生的造血干/祖细胞（HSPC）的影响，以评估祖细胞的适应性和信号传导，以及骨髓肿瘤的长期发展；具体目标 2：我们将鉴定早期骨髓祖细胞中的 Notch 靶基因和表观遗传变化。 DN-MAML1-GFP 融合体上的 GFP 部分将针对染色质免疫沉淀，然后进行 Illumina 双端测序 (ChIP-Seq)，以确定 c-Kit 阳性细胞（富含骨髓干/祖细胞）中 Notch 激活的基因组靶标）。这些研究将与转录分析和额外的 ChIP-Seq 实验相结合，以监测组蛋白修饰。这些研究将定义Notch通路影响骨髓发育的机制，并将确定骨髓特异性Notch功能，这可能最终指导骨髓疾病的新治疗方法。