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.

项目概要/摘要 红细胞生成是一个动态过程，受转录因子相对水平的定量变化控制。 转录因子 (TF)、其特定亚型和翻译后修饰 (PTM)。由于目前缺乏 构成转录调控网络的蛋白质的定量数据，当前的红细胞模型 细胞生成主要基于 mRNA 测量，通常不考虑蛋白质的变化 特定 TF、其异构体或 PTM 的水平。这极大地限制了对红细胞生成和 其他转录调节过程，例如 ss-珠蛋白表达，最终影响 纠正血红蛋白紊乱。长期目标是破译控制红细胞的转录网络 健康和疾病中的细胞生成。该提案的目标是建立红细胞生成的网络模型 基于人类造血红系分化过程中TF蛋白水平的动态变化 干/祖细胞（HSPC）。中心假设是 TF 的相对蛋白质水平是一个关键因素。 在每个分化阶段建立适当的基因表达程序的参数，并且 红细胞生成是由特定转录因子组合的相对量的分级变化驱动的。拉- 关键是将蛋白质组的动态和定量性质整合到转录网络中 红细胞生成的工作将产生一个具有改进预测能力的模型，该模型将作为 健康的红细胞生成与红细胞相关疾病状态进行比较，并将有助于识别 药物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） 根据蛋白质动态变化的测量对红细胞生成转录网络进行建模 转录因子的水平； 2) 识别核蛋白和磷酸原丰度的新变化 红细胞生成过程中的蛋白质。在第一个目标下，开发了一种新颖的靶向质谱方法 申请人将用于测量离体红细胞生成过程中多个阶段的 TF 绝对水平。 HSPC 来自健康捐赠者。在第二个目标下，将使用公正的蛋白质组学方法 识别以前未被重视的蛋白质，这些蛋白质的水平和/或磷酸化发生了定量变化 健康供体的 HSPC 离体红细胞生成期间的状态。该方法具有创新性，因为它 使用新颖的质谱方法来系统地识别和量化调节红细胞生成的转录因子 原代人类细胞中的essis。拟议的研究意义重大，因为它将阐明复杂的监管 控制红细胞生成的过程。最终，这些知识有可能指导新产品的设计 在 SS 地中海贫血患者中重新建立适当的 SS 珠蛋白表达的治疗方法。