The Impact of Nucleolar Stress on Thymocyte Development and T Cell Acute Lymphoblastic Leukemia Transformation

核仁应激对胸腺细胞发育和 T 细胞急性淋巴细胞白血病转化的影响

基本信息

批准号：
9908655
负责人：
JOSEPH RYAN KRAMBS
金额：
$ 3.09万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-09 至 2021-09-08
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9908655
关键词：
Acute Lymphocytic Leukemia Acute T Cell Leukemia Address Adult Binding Biogenesis Bone Marrow CDKN2A gene Cell Line Cell Lineage Cells Cellular Stress Childhood Chimera organism Chimerism Clone Cells Cytoplasm Data Event Feedback Genes Genetic Goals Growth Hematopoietic Hematopoietic Neoplasms Hematopoietic stem cells Induced Mutation Lymphoid MDM2 gene Messenger RNA Modeling Molecular Mus Mutation Myelogenous Myeloproliferative disease NOTCH1 gene Nuclear Pathogenesis Pathway interactions Patients Play Protein Biosynthesis Research Retroviridae Ribosomal Proteins Ribosomes Role Signal Transduction Stem cells Stress T-Cell Development T-Lymphocyte TP53 gene Testing Therapeutic Thymocyte Development Transplantation Xenograft procedure acute T-cell lymphoblastic leukemia cell base c-myc Genes congenic design experimental study gene induction insight knock-down leukemic transformation notch protein novel novel therapeutics progenitor ribosome profiling single-cell RNA sequencing targeted agent transcriptome

项目摘要

Project Summary/Abstract T-ALL represents 10-15% of pediatric and 20-25% of adult cases of acute lymphoblastic leukemia. Activating mutations of NOTCH1 are present in ~60% of cases of T-ALL and are thought to induce leukemic transformation, in part, by activating c-Myc (Myc). Myc induces ribosome biogenesis, which can trigger the nucleolar stress pathway. This is an evolutionarily conserved pathway that includes the translocation of ribosome proteins, RPL5, RPL11, and RPL22 to the nuclear cytoplasm where they bind and alter the activity of p14ARF (ARF) and MDM2, which in turn activates p53, inducing a growth arrest. To overcome this negative feedback loop, over 80% of T- ALL carry biallelic deletions of the CDKN2A (encoding for P16INK4A and ARF). In addition, ~17% of T-ALL carry mutations of RPL5, RPL11, or RPL22. Based on this genetic evidence, we hypothesize that Notch signaling induces nucleolar stress in T-ALL and that inactivation of the nucleolar stress pathway is essential to the molecular pathogenesis of T-ALL. The following specific aims are proposed to test this hypothesis. Aim 1. To determine whether induction of nucleolar stress during normal T-cell development selects for clones carrying mutations in CDKN2A. Notch signaling plays an essential role in normal early T- cell development. We hypothesize that induction of nucleolar stress during early T-cell development selects for T-cell clones carrying mutations of CDKN2A (or RPL genes). To test this hypothesis, we will isolate T-cell progenitors/precursors from wildtype mice and analyze for signs of nucleolar stress. We also will generate Arf or Ink4a deficient mixed bone marrow chimeras and assess donor chimerism. We predict that nucleolar stress in T-cell progenitors/precursors will result in the selective expansion of Arf deficient cells in the T-cell lineage. Aim 2. To determine whether activating Notch1 mutations induce nucleolar stress in T cell progenitors. We will assess the impact of the enforced expression of NOTCH1 mutations found in T-ALL on ribosome biogenesis and induction of nucleolar stress using the MOLM13 cell line and primary murine T-cells. In parallel studies, we also will knock-down NOTCH1 expression in several T-ALL xenografts. In each case, nucleolar stress will be assessed

项目摘要/摘要 T-ALL代表10-15％的小儿和20-25％的成年急性淋巴细胞白血病病例。激活 Notch1的突变存在于〜60％的T-ALL病例中，被认为会诱导白血病转化，部分通过激活C-MYC（MYC）。 MYC诱导核糖体生物发生，可以触发核仁应力路径。这是一种进化保守的途径，包括核糖体蛋白Rpl5， RPL11和RPL22到核细胞质，它们结合并改变了p14arf（ARF）和MDM2的活性，这反过来激活了p53，引起了生长停滞。为了克服这种负反馈循环，超过80％的t- 所有人都带有CDKN2A的双重缺失（编码P16INK4A和ARF）。此外，约有17％的T-All随身携带 RPL5，RPL11或RPL22的突变。基于此遗传证据，我们假设Notch信号传导诱导T-ALL诱导核仁应力，并且核仁应力途径的失活对于 T-ALL的分子发病机理。提出了以下特定目的来检验这一假设。目的1。确定在正常T细胞发育过程中是否诱导核仁应力选择在CDKN2A中携带突变的克隆。 Notch信号在正常的早期T-中起着至关重要的作用细胞发育。我们假设在早期T细胞发育过程中诱导核仁应力会选择带有CDKN2A突变（或RPL基因）突变的T细胞克隆。为了检验这一假设，我们将隔离T细胞野生型小鼠的祖/前体，分析核仁应力的迹象。我们还将生成ARF 或Ink4a缺乏混合骨髓嵌合体并评估供体嵌合体。我们预测核仁应力在T细胞祖细胞/前体中，将导致T细胞谱系中ARF缺陷细胞的选择性扩展。目标2。确定激活Notch1突变是否诱导T细胞中的核仁应力祖先。我们将评估T-All中发现Notch1突变的强制表达的影响核糖体生物发生和使用Molm13细胞系和原代鼠T细胞诱导核仁应力。在平行研究中，我们还将在几个T-All异种移植物中敲低Notch1的表达。在每种情况下，核仁应力将被评估