Transcriptional Control During Erythropoiesis

红细胞生成过程中的转录控制

• 批准号: 8734411
• 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/8734411
• 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

DESCRIPTION (provided by applicant): Erythropoiesis is a dynamic process governed by quantitative changes in the relative levels of transcription factors (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 erythropoiesis 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 th understanding of erythropoiesis and other transcriptionally regulated processes such as ¿-globin expression, ultimately impinging on the capacity to correct hemoglobin disorders. The long-term goal is to decipher the transcriptional network that controls erythropoiesis 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 parameter 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 rationale is that integration of the dynamic and quantitative nature of the proteome into the transcriptional network 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 identification o 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 phosphoproteins 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 phosphorylation 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 erythropoiesis 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 ¿-globin expression in ¿-thalassemic patients.

描述（由申请人提供）：红细胞生成是一个动态过程，由转录因子（TF）及其特异性亚型的相对水平的定量变化控制 和翻译后修饰（PTM）。由于目前缺乏关于构成转录调控网络的蛋白质的定量数据，目前的红细胞生成模型主要基于mRNA测量，并且通常不考虑特定TF、其同种型或PTM的蛋白质水平的变化。这极大地限制了对红细胞生成和其他转录调控过程（如β-珠蛋白表达）的理解，最终影响了纠正血红蛋白紊乱的能力。长期目标是破译控制健康和疾病中红细胞生成的转录网络。 本研究的目的是建立一个基于人造血干/祖细胞（HSPCs）向红系分化过程中TF蛋白水平动态变化的红细胞生成网络模型。中心假设是TF的相对蛋白水平是 在每个分化阶段建立适当的基因表达程序的关键参数，并且红细胞生成由TF的特定组合的相对量的分级变化驱动。其基本原理是将蛋白质组的动态和定量性质整合到红细胞生成的转录网络中将产生具有改善的预测能力的模型，其将用作健康红细胞生成的基准，与之比较红细胞相关的疾病状态，并将有助于鉴定恢复正常红细胞生成的药理学试剂。已经设计了两个具体目标：1）基于转录因子的蛋白质水平的动态变化的测量来对红细胞生成转录网络进行建模;以及2）鉴定红细胞生成期间核蛋白和磷蛋白丰度的新变化。在第一个目标下，由申请人开发的新型靶向质谱法将用于测量源自健康供体的HSPC的离体红细胞生成期间多个阶段的TF的绝对水平。在第二个目标下，将使用无偏倚的蛋白质组学方法来鉴定在来自健康供体的HSPC的离体红细胞生成期间经历其水平和/或磷酸化状态的定量变化的先前未被认识的蛋白质。该方法是创新的，因为它使用新的质谱方法来系统地鉴定和定量调节原代人类细胞中红细胞生成的TF。这项研究意义重大，因为它将阐明控制红细胞生成的复杂调控过程。最终，这些知识有可能指导新疗法的设计，以在地中海贫血患者中重新建立适当的珠蛋白表达。