Function of RUNX1 in diverse Down syndrome tissues

RUNX1在多种唐氏综合症组织中的功能

• 批准号: 10853906
• 负责人: Mary A Allen
• 金额: $ 19.15万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10853906
• 关键词: Address; Administrative Supplement; Adult; Affect; Alternative Splicing; Award; Basic Science; Binding; Biological Assay; Blood; Blood Cells; Cell Differentiation process; Cell Line; Cells; Chromatin; Chromatin Remodeling Factor; Chromosome 21; DNA; DNA Binding; DNA Binding Domain; DNA-Directed RNA Polymerase; Data; Defect; Development; Disease; Down Syndrome; Embryonic Development; Equilibrium; GATA1 gene; Gene Dosage; Genetic Transcription; Grant; Hematological Disease; Hematology; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; High Prevalence; Histones; Homeostasis; Incidence; Individual; Knock-out; Life; Link; Mediating; Megakaryocytopoieses; Methods; Molecular; Motivation; Mutation; Myeloid Leukemia; Outcome; Parents; Pattern; Pharmaceutical Preparations; Phenotype; Play; Protein Isoforms; RNA Polymerase II; RNA Splicing; RUNX1 gene; Recruitment Activity; Reporting; Risk; Role; Specific qualifier value; Tissues; Transactivation; Transcription Coactivator; Trisomy; Variant; Work; acute myeloid leukemia 1 protein; blastomere structure; cell type; cost; dosage; embryo cell; high reward; high risk; improved; induced pluripotent stem cell; insight; leukemia; loss of function mutation; novel; overexpression; postnatal; recruit; transcription factor

Project Summary Parent Proposal: RUNX1, also known as Acute Myeloid Leukemia 1 protein (AML1), is a transcription factor that plays a critical function in the specification of the hematopoietic lineage during embryogenesis and is required for normal megakaryopoiesis throughout the postnatal life. The RUNX1 gene is localized to band q22.12 of chromosome 21, which is triplicated in individuals with Trisomy 21 (T21). For this reason, an extra copy of RUNX1 has been proposed to play relevant roles in the hematological alterations associated with Down syndrome (DS). We seek to decipher the contribution of RUNX1 to the phenotypes seen in individuals with Down syndrome. Since transcription factors bind to DNA and alter RNA polymerase activity, we will determine if those two functions of RUNX1 are altered in Down syndrome-derived cells. We will also define if drugs that increase or inhibit RUNX1 function behave differently in cells with an extra copy of chromosome 21. Finally, we will ascertain how much of the altered blood differentiation seen in Down syndrome is caused by RUNX1 by ‘normalizing’ RUNX1 gene dosage in a trisomy background (e.g., two copies in a trisomy cell line). We will then analyze the differentiation of iPSCs into embryoid bodies and blood cells. Collectively this work will shed important insights into the functions of RUNX1 and how it is altered in Trisomy 21. Supplement Project Summary: A recent report by Gialesaki et al. (March 2023) established a critical role for RUNX1 alternative splicing and isoform imbalance -rather than overall expression- as a major driver of Down syndrome-associated myeloid leukemia (DS-ML). These findings have important implications on most of the specific aims of our proposal. Therefore, we should partially modify some of our sub-aims to assess the relevance of RUNX1 splicing equilibrium on the different outcomes of DS-derived iPSC and lymphoblastoid cell differentiation and homeostasis. A second motivation for this supplement applies exclusively to Aim 3. Here we orginally proposed to normalize RUNX1 gene dosage by knocking-out one copy in the T21 +/+/− iPSC line (T21 RUNX1 ). We now propose alternative strategies to overcome such obstacles and incorporate the isoform-specific analyses just mentioned. This Administrative Supplement addresses Component 1: A targeted high risk - high reward basic science study that is relevant to Leukemia risk in Down syndrome. Moreover this supplement will significantly improve the scientific significance of the project by contributing to a better understanding of the underlying mechanisms that govern hematological alterations in individuals with T21.

项目摘要 家长建议：runx1，也称为急性髓性白血病1蛋白（AML1）是一个 在造血谱系规范中起关键功能起着关键功能的转录因子 在胚胎发生过程中，在整个后期中需要正常的巨型脑膜 生活。 Runx1基因本地化为21染色体的Q22.12，该基因在 三体术21（T21）。因此，已经提出了额外的runx1副本 在与唐氏综合症（DS）相关的血液学改变中起相关的作用。我们 寻求破译Runx1对有降低个人的表型的贡献 综合征。由于转录因子与DNA结合并改变RNA聚合酶活性，我们将 确定Runx1的这两个功能是否在唐氏综合征衍生的细胞中改变。我们将 还定义增加或抑制RUNX1功能的药物在具有的细胞中的表现不同 染色体21的额外副本。最后，我们将确定多少鲜血 唐氏综合症中看到的差异化是由runx1通过“归一化” runx1基因引起的 三体背景中的剂量（例如，三体细胞系中的两个副本）。然后，我们将分析 IPSC分化为胚胎体和血细胞。总的来说这项工作将丢弃 对Runx1功能的重要见解及其如何改变三体性。 补充项目摘要：Gialesaki等人的最新报告。 （2023年3月）建立了 Runx1替代剪接和同工型不平衡的关键作用，而不是整体 表达 - 作为唐氏综合症相关的髓样白血病（DS-ML）的主要驱动力。这些 发现对我们提案的大多数特定目的具有重要意义。因此，我们 是否应该部分修改我们的一些子iams来评估Runx1剪接的相关性 DS衍生的IPSC和淋巴母细胞分化的不同结果平衡 和稳态。该补充剂的第二个动机仅适用于目标3。 我们通常建议通过在T21中淘汰一个副本来标准化Runx1基因剂量 +/+/ - IPSC线（T21 Runx1）。我们现在提出了克服此类的替代策略 障碍并结合了刚刚提到的同工型特异性分析。这个行政 补充解决方案1：有针对性的高风险 - 高奖励基础科学研究 这与唐氏综合症的白血病风险有关。此外，这种补充将大大 通过更好地了解该项目的科学意义 控制T21个体血液学改变的基本机制。