EpoR & Stat5 regulation of ribosome biogenesis and protein synthesis in erythropoiesis

EPR

基本信息

批准号：
10682214
负责人：
Merav Socolovsky
金额：
$ 51.52万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-05-01 至 2028-02-29
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10682214
关键词：
Address Back Biogenesis Biomass Bone Marrow CFU-E Cell Cycle Cell Enlargement Cell Lineage Cell Size Cell division Cells Compensation Complex Cues Cytokine Receptors Data Development Diamond-Blackfan anemia Disease Embryo Erythroblasts Erythrocytes Erythroid Erythropoiesis Erythropoietin FRAP1 gene Fetal Liver Genes Genetic Genetic Transcription Growth Growth Factor Hematopoiesis Hemoglobin Hormones Housekeeping Human Insulin Receptor Knowledge Life Link Malignant Neoplasms Mammalian Cell Measures Mediating Mediator Modeling Mus Mutation Nutritional PIK3CG gene Pathway interactions Phosphotransferases Process Protein Biosynthesis Protein Synthesis Induction Proteins Proteome Proto-Oncogene Proteins c-myc Publishing Receptor Protein-Tyrosine Kinases Regulation Ribosomal Proteins Ribosomal RNA Ribosomes S phase Signal Pathway Signal Transduction Speed Testing Time Transcript Translating Translations Work attenuation erythroid differentiation interest member novel overexpression pharmacologic pressure progenitor ribosomopathy self-renewal single-cell RNA sequencing transcriptomics

项目摘要

Project Summary Protein synthesis and ribosome biogenesis are the two most energy -intensive processes in a cell, resulting in their regulation through tight controls. EpoR, a member of the cytokine receptor superfamily, and its downstream mediator Stat5, are essential for erythropoiesis. The current proposal addresses novel regulation of ribosome biogenesis and protein synthesis by EpoR and Stat5 signaling during a key erythroid developmental decision. In recently published work we found that EpoR/ Stat5 stimulate shorter and more numerous cycles in early erythroblasts, while also promoting formation of larger erythroblasts that mature into larger red cells, in both mice and humans. This surprising finding suggests that EpoR/ Stat5 signaling alters the relationship between cell cycle duration and cell size, simultaneously inducing shorter cycles and exceptionally fast growth in biomass. In preliminary data supporting this hypothesis, single-cell RNA sequencing of Epor-/- and Stat5-/- fetal livers show dysregulated expression of ribosome biogenesis and translation genes. Further, we identified an EpoR/ Stat5-dependent sharp spike in the rate of rRNA transcription (~2.5 fold), protein synthesis rate (~4 to 6 fold) and rate of growth in cell size (~3 fold), that coincides with a key cell fate decision. It takes place as early erythroid progenitors known as CFU-e transition from self-renewal to erythroid terminal differentiation (ETD), becoming erythroblasts. The spike in protein synthesis at this time also coincides with an unusually short cell cycle. Immediately following the CFU-e/ETD transition, the rates of ribosome biogenesis and protein synthesis begin to decline back to baseline together with the decline in cell cycle speed, even though erythroblasts continue to divide and synthesize hemoglobin for an additional 3 to 5 cell divisions. The coincidence in the spikes of protein synthesis and ribosome biogenesis with cell cycle shortening at the CFU-e/ETD switch suggests that these processes are linked and may be functionally relevant to the switch. We will investigate the EpoR/Stat5- induced spike in protein synthesis with the following aims: Aim 1: Investigate the EpoR /Stat5 -induced spike in ribosome biogenesis & protein synthesis, determining the intracellular signaling pathways that are mediating this spike and identifying a potential subset of transcripts whose translation rate spikes. Aim 2: Determine causal relationships between the cell cycle, ribosome biogenesis, protein synthesis and cell size. The regulatory interactions between cell cycle duration, protein synthesis rate and cell size in mammalian cells are not well understood and yet are critical in development and in cancer. Aim 3: Test the hypothesis that the EpoR /Stat5 -induced spike in protein synthesis is required for erythroid differentiation. We will determine whether the EpoR-induced spike in protein synthesis is an 'Achilles heel' in mice deficient in the ribosomal protein Rpl11, a model of Diamond Blackfan Anemia, potentially explaining the selective sensitivity of the erythroid lineage to ribosomopathies.

项目概要蛋白质合成和核糖体生物发生是两个最耗能的领域细胞中的过程，通过严格的控制对其进行调节。 EpoR，细胞因子受体的成员超家族及其下游介质 Stat5 对于红细胞生成至关重要。目前的提案解决了 EpoR 和 Stat5 信号传导对核糖体生物发生和蛋白质合成的新调节关键的红细胞发育决定。在最近发表的工作中，我们发现 EpoR/ Stat5 刺激时间更短以及早期有红细胞中更多的循环，同时还促进更大的有红细胞的形成在小鼠和人类中，成熟为更大的红细胞。这一令人惊讶的发现表明 EpoR/Stat5 信号传导改变细胞周期持续时间和细胞大小之间的关系，同时诱导更短的细胞周期循环和生物量异常快速的增长。在支持这一假设的初步数据中，单细胞 RNA Epor-/- 和 Stat5-/- 胎儿肝脏测序显示核糖体生物发生和表达失调翻译基因。此外，我们还发现了 rRNA 速率依赖于 EpoR/Stat5 的急剧上升。转录（约 2.5 倍）、蛋白质合成速率（约 4 至 6 倍）和细胞大小生长速率（约 3 倍），与关键的细胞命运决定相一致。它作为早期红系祖细胞发生，称为 CFU-e 转变从自我更新到红细胞终末分化（ETD），成为红细胞。蛋白质含量激增此时的合成也与异常短的细胞周期相一致。紧接着 CFU-e/ETD 转变时，核糖体生物合成和蛋白质合成的速率开始一起下降回到基线随着细胞周期速度的下降，即使有红细胞继续分裂和合成血红蛋白额外 3 至 5 次细胞分裂。蛋白质合成和核糖体尖峰的重合 CFU-e/ETD 开关处细胞周期缩短的生物发生表明这些过程是相互关联的并且可能与开关功能相关。我们将研究 EpoR/Stat5 诱导的蛋白质尖峰合成具有以下目标：目标 1：研究 EpoR /Stat5 诱导的核糖体生物发生中的尖峰 & 蛋白质合成，确定介导这种尖峰的细胞内信号传导途径并识别翻译率飙升的潜在转录本子集。目标 2：确定之间的因果关系细胞周期、核糖体生物合成、蛋白质合成和细胞大小。细胞间的调控相互作用哺乳动物细胞的周期持续时间、蛋白质合成速率和细胞大小尚未得到充分了解，但对发育和癌症至关重要。目标 3：检验 EpoR /Stat5 诱导蛋白质峰值的假设红细胞分化需要合成。我们将确定 EpoR 是否诱导蛋白质尖峰合成是缺乏核糖体蛋白 Rpl11（Diamond Blackfan 模型）的小鼠的“致命弱点” 贫血，可能解释红系谱系对核糖体病的选择性敏感性。