Contribution of Zranb3 to normal and stressed hematopoiesis

Zranb3 对正常和应激造血的贡献

• 批准号: 10668267
• 负责人: Saul Kushinsky
• 金额: $ 5.27万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10668267
• 关键词: Acceleration; Apoptosis; B-Cell Lymphomas; Biochemical; Biological; Blood; Blood Cells; Bone Marrow; Bone Marrow Cells; Bone marrow failure; CD34 gene; Cancer cell line; Cell Death; Cells; Cellular Stress; Cessation of life; DNA; DNA Damage; DNA Sequence Alteration; DNA biosynthesis; DNA replication fork; DNA strand break; Data; Defect; Development; Doctor of Philosophy; Education; Emergency Situation; Evaluation; Failure; Family member; Genes; Genome; Genomic Instability; Goals; Hematology; Hematopoiesis; Hematopoietic; Hematopoietic System; Hematopoietic stem cells; Human; Investigation; Knowledge; Life; Lymphoid; Mentors; Mus; Myelogenous; Pathway interactions; Physicians; Physiological; Population; Process; Proliferating; Proteins; Reporting; Role; Scientist; Source; Stem Cell Development; Stimulus; Stress; Testing; Training; Universities; biological adaptation to stress; cancer cell; career development; defined contribution; experimental study; gene conservation; in vivo; in vivo Model; insight; knock-down; mouse model; preference; preservation; prevent; programs; protective effect; repaired; replication stress; response; skills; stem cell function; stem cell proliferation; transcriptome sequencing

Project Summary Hematopoiesis, the process by which hematopoietic stem and progenitor cells (HSPCs) in the bone marrow produce all mature blood cells for the hematopoietic system, is required for life. HSPCs are particularly prone to DNA replication stress due to their high rates of replication during normal hematopoiesis and certainly during bursts of proliferation from stress-induced/emergency hematopoiesis. DNA replication stress that is not properly resolved is postulated to contribute to genomic instability and cell death, ultimately leading to HSPC defects, deficiencies and/or bone marrow failure. Yet, the proteins that mitigate such stress in HSPCs remain largely uncharacterized. Understanding the proteins that respond to DNA replication stress in HSPCs is essential to understanding how HSPC development and function is protected throughout a lifetime. Recently, Smarcal1 and Zranb3, two proteins with the same biochemical activity of fork reversal and remodeling, were determined in human cancer cells to respond to DNA replication stress and prevent DNA replication fork collapse, but their role in vivo and in hematopoietic cells remained unexplored. The Eischen lab discovered that while Smarcal1 is not required for normal hematopoiesis, it is essential for HSPCs during stressed/emergency hematopoiesis. Currently, little is known about Zranb3 in vivo, as well as whether the functions of Smarcal1 and Zranb3 are redundant in vivo in hematopoietic cells. Our preliminary data suggest Zranb3 may be essential for HSPCs and its functions are non-redundant with Smarcal1 in vivo. Therefore, we hypothesize Zranb3 is an essential component of the DNA replication stress response in HSPCs and significantly contributes to their proliferation and survival. To test this hypothesis, we will use mouse models and both mouse and human primary hematopoietic cells. In Aim 1, we propose to investigate the requirements of Zranb3 to normal hematopoiesis and the mechanism involved. In Aim 2, we will investigate the contribution of Zranb3 to DNA replication stress- induced during stressed/emergency hematopoiesis. Results from the proposed experiments will significantly increase understanding of the DNA replication stress response that protects hematopoietic cells during normal and stressed hematopoiesis and prevents hematopoietic cell failure. The MD/PhD program at Thomas Jefferson University provides challenging, comprehensive training that will allow me to fulfill the goals of this application and become a successful independent physician-scientist. My training will include the development of many skills and an increase of knowledge through multiple approaches supported by my mentor, thesis committee, and other scientists. The educational and career development objectives outlined in this proposal will help me fulfill my goal of becoming a successful, independent physician- scientist focused on hematology.

项目摘要 造血，骨髓中的造血干细胞和祖细胞（HSPC） 产生造血系统所需的所有成熟血细胞，是生命所必需的。HSPC特别容易出现 DNA复制应激是由于它们在正常造血过程中的高复制率， 应激诱导/紧急造血引起的增殖爆发。DNA复制压力， 假定分解有助于基因组不稳定性和细胞死亡，最终导致HSPC缺陷， 缺乏和/或骨髓衰竭。然而，在HSPCs中减轻这种压力的蛋白质在很大程度上仍然存在， 没有特征的了解HSPCs中响应DNA复制应激的蛋白质对于 了解HSPC的发育和功能如何在整个生命周期内得到保护。最近，Smarcal 1和 Zranb 3是两种具有相同的叉逆转和重构生化活性的蛋白质， 人类癌细胞对DNA复制应激作出反应，防止DNA复制叉崩溃，但它们的作用 在体内和造血细胞中的作用尚未探索。科布琴实验室发现，虽然Smarcal 1不是 正常造血所需，在应激/紧急造血期间对HSPC是必需的。 目前，对体内的Zranb 3以及Smarcal 1和Zranb 3的功能是否被激活知之甚少。 在体内造血细胞中是多余的。我们的初步数据表明，Zranb 3可能是HSPC所必需的， 其功能与体内Smarcal 1无冗余。因此，我们假设Zranb 3是一个重要的 HSPCs中DNA复制应激反应的组成部分，并显著促进其增殖 和生存为了验证这一假设，我们将使用小鼠模型， 造血细胞在目的1中，我们提出研究Zranb 3对正常造血的需求 以及所涉及的机制。在目标2中，我们将研究Zranb 3对DNA复制应激的贡献- 在应激/紧急造血期间诱导。从拟议的实验结果将显著 增加对DNA复制应激反应的理解，该反应在正常情况下保护造血细胞。 和防止造血细胞衰竭。 托马斯杰斐逊大学的MD/PhD课程提供具有挑战性的全面培训， 将使我能够实现这个应用程序的目标，并成为一个成功的独立的物理学家科学家。我 培训将包括通过多种方法发展多种技能和增加知识 得到了我的导师论文委员会和其他科学家的支持教育和职业发展 本计划书中列出的目标将帮助我实现成为一名成功的独立医生的目标。 专注于血液学的科学家