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.

项目概要 家长提案：RUNX1，也称为急性髓系白血病 1 蛋白 (AML1)，是一种 在造血谱系规范中发挥关键作用的转录因子 在胚胎发生期间，是整个出生后正常巨核细胞生成所必需的 RUNX1 基因定位于 21 号染色体的带 q22.12，该带在 21 号染色体中是三倍的。 因此，建议为 21 三体 (T21) 患者提供额外的 RUNX1 副本。 在与唐氏综合症（DS）相关的血液学改变中发挥相关作用。 试图破译 RUNX1 对唐氏综合症患者表型的贡献 由于转录因子与 DNA 结合并改变 RNA 聚合酶活性，我们将 我们将确定 RUNX1 的这两个功能是否在唐氏综合症衍生细胞中发生改变。 还定义了增加或抑制 RUNX1 功能的药物在具有 RUNX1 功能的细胞中是否表现不同 21号染色体的额外拷贝。最后，我们将确定有多少被改变的血液 唐氏综合症中看到的分化是由 RUNX1 通过“正常化”RUNX1 基因引起的 然后我们将分析三体背景中的剂量（例如，三体细胞系中的两个拷贝）。 iPSC 分化为胚状体和血细胞。 对 RUNX1 功能及其在 21 三体中如何改变的重要见解。 补充项目摘要：Gialesaki 等人最近的一份报告（2023 年 3 月）建立了 RUNX1 选择性剪接和异构体不平衡的关键作用 - 而不是整体 表达-作为唐氏综合症相关骨髓性白血病（DS-ML）的主要驱动因素。 研究结果对我们提案的大多数具体目标具有重要意义。 应该部分修改我们的一些子目标来评估 RUNX1 剪接的相关性 DS 来源的 iPSC 和类淋巴母细胞分化的不同结果的平衡 这种补充剂的第二个动机仅适用于目标 3。 我们最初提出通过敲除 T21 中的一个拷贝来标准化 RUNX1 基因剂量 +/+/− iPSC 系 (T21 RUNX1) 我们现在提出替代策略来克服此类问题。 障碍并纳入刚才提到的异构体特异性分析。 补充内容涉及第 1 部分：有针对性的高风险高回报基础科学研究 这与唐氏综合症的白血病风险有关。 通过促进更好地理解项目来提高项目的科学意义 控制 T21 个体血液学改变的潜在机制。

项目成果

Characterizing Primary 1 transcriptional responses to short 2 term heat shock in paired fraternal 3 lymphoblastoid lines with and 4 without Down syndrome.

描述患有唐氏综合症和不患有唐氏综合症的配对兄弟 3 个淋巴母细胞系中对短期 2 期热休克的初级 1 转录反应。

• 发表时间: 2024-09-14
• 作者: Joseph F. Cardiello;Jessica Westfall;Robin Dowell;Mary Ann Allen
• 通讯作者: Mary Ann Allen