Transcriptional Control During Erythropoiesis

红细胞生成过程中的转录控制

• 批准号: 8881162
• 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 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8881162
• 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; 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（由应用程序提供）：红细胞生成是一个动态过程，由转录因子（TFS）的相对水平（TFS）的定量变化（其特定的同工型）控制 和翻译后修改（PTMS）。由于目前缺乏构成转录调节网络的蛋白质的定量数据，因此，红细胞生成的当前模型主要基于mRNA测量，并且通常不考虑特定TFS蛋白质水平的变化，其同工型或PTMS。这显着限制了对促红细胞生成的理解和其他转录调节的过程，例如``全球剂表达''，最终影响了纠正血红蛋白疾病的能力。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。 该提案的目的是基于人类造血干/祖细胞（HSPCS）的红细胞分化过程中TF蛋白水平的动态变化，建立红细胞生成的网络模型。中心假设是TF的相对蛋白水平是 在分化的每个阶段建立适当的基因表达程序的关键参数，并且红细胞生成是由TFS特定组合的相对量的分级变化驱动的。理由是，将蛋白质组的动态和定量性质的整合到促红细胞生成的转录网络将导致具有提高预测能力的模型，这将是健康的红细胞生成基准，以使其与核苷相关疾病状态的健康性促红细胞生成，并将促进识别型e Pharbiace o pharmaceporical eRy ersore erey ery ere hory ery hory ery ery ery him y himory ery。已经设计了两个具体目标：1）基于转录因子蛋白质水平的动态变化的测量，对红细胞生成的转录网络进行了建模； 2）确定红细胞生成期间核蛋白和磷蛋白抽象的新变化。在第一个目标下，申请人开发的一种新型的靶向质谱方法将用于测量来自健康供体的HSPC的离体红细胞生成期间多个阶段的绝对水平。在第二个目标下，将使用一种公正的蛋白质组学方法来识别先前未经批准的蛋白质，该蛋白质会在健康供体的HSPC中发生定量变化和/或磷酸化状态的定量变化和/或磷酸化状态。该方法具有创新性，因为它使用新型的质谱方法来系统地识别和量化调节原代人类细胞中红细胞生成的TF。拟议的研究很重要，因为它将照亮控制红细胞生成的复杂调节过程。最终，这种知识有可能指导新的治疗剂的设计，以重新建立thalassemic患者的适当 - 全珠蛋白表达。