Transcriptional Control During Erythropoiesis

红细胞生成过程中的转录控制

• 批准号: 8632907
• 负责人: Marjorie Carole Brand
• 金额: $ 31.32万
• 依托单位: INSTITUTE FOR SYSTEMS BIOLOGY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-16 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8632907
• 关键词: Adult; Benchmarking; Binding; Bone Marrow; Cell physiology; Cells; Complex; Computer Simulation; Data; Development; Disease; Ensure; Erythrocytes; Erythroid; Erythroid Cells; Erythropoiesis; GATA1 gene; Gene Expression; Genes; Genetic Transcription; Genomics; Globin; Goals; Health; Hematopoietic; Hematopoietic stem cells; Hemoglobin; Human; Knowledge; Lead; Mass Spectrum Analysis; Measurement; Measures; Messenger RNA; Methods; Mission; Modeling; Mutation; Nature; Network-based; Nuclear Proteins; Outcome; Patients; Peptides; Phosphoproteins; Phosphorylation; Post-Translational Protein Processing; Process; Production; Protein Isoforms; Proteins; Proteome; Proteomics; Proxy; Public Health; Regulation; Relative (related person); Research; Role; Sampling; Stable Isotope Labeling; Staging; Stem cells; Techniques; Technology; Thalassemia; Time; Transcriptional Regulation; Umbilical Cord Blood; Work; base; comparative; design; dosage; erythroid differentiation; fetal; genome-wide; improved; innovation; insight; mRNA Expression; network models; novel; novel therapeutics; programs; public health relevance; research study; stem; transcription factor

PROJECT SUMMARY/ABSTRACT Erythropoiesis is a dynamic process governed by quantitative changes in the relative levels of transcription fac- tors (TFs), their specific isoforms and post-translational modifications (PTMs). Due to the current paucity of quantitative data on the proteins that constitute the transcriptional regulatory network, current models of eryth- ropoiesis are based primarily on mRNA measurements and do not typically consider changes in the protein levels of specific TFs, their isoforms or PTMs. This significantly limits the understanding of erythropoiesis and other transcriptionally regulated processes such as ss-globin expression, ultimately impinging on the capacity to correct hemoglobin disorders. The long-term goal is to decipher the transcriptional network that controls eryth- ropoiesis in health and disease. The objective of this proposal is to build a network model of erythropoiesis based on dynamic changes in TF protein levels during erythroid differentiation of human hematopoietic stem/progenitor cells (HSPCs). The central hypothesis is that the relative protein levels of TFs is a critical pa- rameter in the establishment of proper gene expression programs at each stage of differentiation, and that erythropoiesis is driven by graded changes in the relative amounts of specific combinations of TFs. The ra- tionale is that integration of the dynamic and quantitative nature of the proteome into the transcriptional net- work of erythropoiesis will result in a model with improved predictive power which will serve as a benchmark for healthy erythropoiesis against which to compare erythroid-related disease states, and will facilitate the identifi- cation of pharmacological agents to restore normal erythropoiesis. Two specific aims have been designed: 1) Model the erythropoiesis transcriptional network based on measurements of dynamic changes in the protein levels of transcription factors; and 2) Identify novel changes in abundance of nuclear proteins and phosphopro- teins during erythropoiesis. Under the first aim, a novel targeted mass spectrometry approach developed by the applicants will be used to measure absolute levels of TFs at multiple stages during ex vivo erythropoiesis of HSPCs derived from healthy donors. Under the second aim, an unbiased proteomic approach will be used to identify previously unappreciated proteins that undergo quantitative changes in their levels and/or phosphoryla- tion status during ex vivo erythropoiesis of HSPCs from healthy donors. The approach is innovative because it uses novel mass spectrometry approaches to systematically identify and quantify TFs that regulate erythropoi- esis in primary human cells. The proposed research is significant because it will illuminate complex regulatory processes that control erythropoiesis. Ultimately, such knowledge has the potential to guide the design of new therapeutics to re-establish proper ss-globin expression in ss-thalassemic patients.

项目摘要/摘要 红细胞生成是一个动态过程，该过程受转录相对水平的定量变化的控制 TOR（TFS），其特定的同工型和翻译后修饰（PTMS）。由于目前的稀少 关于构成转录调节网络的蛋白质的定量数据 ropoiesis主要基于mRNA测量，通常不考虑蛋白质的变化 特定TF的水平，它们的同工型或PTM。这显着限制了对促红细胞生成和的理解 其他转录调节的过程，例如SS-珠蛋白的表达，最终影响 纠正血红蛋白疾病。长期目标是破译控制生成的转录网络 健康和疾病中的ropoiesis。该提案的目的是建立红细胞生成的网络模型 基于人造血的红细胞分化期间TF蛋白水平的动态变化 茎/祖细胞（HSPC）。中心假设是TFS的相对蛋白水平是关键的PA- Rameter在分化的每个阶段建立适当的基因表达程序，并 红细胞生成是由TFS特定组合的相对量的分级变化驱动的。 ra- tionale是蛋白质组的动态和定量性质的整合到转录净的 红细胞生成的工作将导致具有提高预测能力的模型，这将作为基准 健康的红细胞生成，以比较与红细胞相关疾病状态的状态，并将促进鉴定 药理学剂的阳离子以恢复正常的红细胞生成。设计了两个具体目标：1） 基于蛋白质动态变化的测量，建模红细胞生成的转录网络 转录因子的水平； 2）确定核蛋白丰富和磷酸化的新变化 红细胞生成期间的Teins。在第一个目标下，一种新型的针对质谱方法由 申请人将用于测量在体内促红细胞生成期间多个阶段的绝对水平 HSPC来自健康的捐助者。在第二个目标下，将使用公正的蛋白质组学方法 确定先前未批准的蛋白质，这些蛋白质会在其水平和/或磷酸化的水平上发生定量变化 来自健康捐助者的HSPC的离体红细胞生成期间的状态。这种方法是创新的，因为它 使用新型的质谱法系统识别和量化调节红细胞生成的TFS 原代人细胞中的ESI。拟议的研究很重要，因为它将照亮复杂的调节 控制红血病的过程。最终，这种知识有可能指导新的设计 在SS-丘脑血症患者中重新建立适当的SS-球蛋白表达的治疗剂。