The Role of MYC Mutations in acute Myeloid Leukemia

MYC 突变在急性髓系白血病中的作用

• 批准号: 10656311
• 负责人: Francesca Ferraro
• 金额: $ 22.5万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-13 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10656311
• 关键词: Acute Myelocytic Leukemia; Address; Affect; Alleles; B lymphoid malignancy; Binding Sites; Breeding; Candidate Disease Gene; Cells; ChIP-seq; Characteristics; Chromosome 8; DNA Binding; Data Analyses; Data Set; Development; Development Plans; Diagnosis; Disease remission; Dose; Doxycycline; Frequencies; Future; Gene Expression Profile; Genes; Genetic Transcription; Genomics; Goals; Growth; Half-Life; Hematopoietic; Hematopoietic stem cells; In Vitro; Karyotype; Knock-in; Knowledge; Leukemic Cell; MYC Box; MYC Family Protein; Mediating; Missense Mutation; Modeling; Molecular; Mus; Mutation; NPM1 gene; Oncogenic; Outcome; Output; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Phenotype; Physiological; Property; Proteins; RNA Splicing; Recurrence; Research Project Grants; Resistance; Resources; Risk; Role; Sampling; Series; Solid Neoplasm; System; Technology; Translations; Universities; Washington; Work; acute myeloid leukemia cell; candidate identification; career; career development; cell transformation; chemotherapy; cohort; design; differential expression; effectiveness evaluation; experimental study; improved; improved outcome; in vivo; insight; leukemia; leukemic transformation; leukemogenesis; medical schools; mouse model; mutant; novel strategies; novel therapeutic intervention; overexpression; predictive tools; research and development; response; risk stratification; single-cell RNA sequencing; skills; treatment response

Project Summary This proposal outlines a 5-year research and career development plan designed to support the candidate’s trajectory towards an independent academic career. The proposed research project, which focuses on acute myeloid leukemia (AML)-associated MYC mutations and their key target genes in AML cells, will capitalize on critical expertise and resources available at Washington University School of Medicine. The career development plan and didactic work will provide the candidate with a variety of skills that will enable a successful transition to independence. This proposal is founded on our recent observation that a unique set of recurrent MYC missense mutations are significantly enriched in the dominant clones of normal karyotype AML patients who have very long first remissions (>5 years) with chemotherapy only; in our series, 6/6 MYC mutations co- occurred with NPM1 mutations (NPMc), suggesting a unique form of cooperativity. Based on our preliminary findings, we hypothesize that: 1) AMLs overexpressing WT or mutant MYC proteins are biologically different; 2) each MYC mutation may have unique effects on its transcriptional targets; and 3) MYC mutations may cooperate with NPMc mutations. To address these hypotheses, we propose the following specific aims: Specific Aim 1: We will examine the function of AML-associated MYC mutations alone, or in cooperation with NPMc. We have developed a doxycycline-inducible system that allows us to regulate Myc expression in hematopoietic stem and progenitor cells (HSPCs) that do or do not contain the Npmc mutation. Using this system, we will evaluate how WT or mutant Myc, with or without Npmc, impacts AML development, progression, and AraC responsiveness both in vitro and in vivo. We are also developing a more physiologic, conditional knock-in model of MYCT58N (a recurrent mutation both in AML and B cell malignancies) that will allow us to explore the influence of physiologic doses of mutant Myc and Npmc on AML pathogenesis and AraC sensitivity. Specific Aim 2: We will define the transcriptional outputs and genomic binding sites of WT vs. mutant Myc proteins in AMLs arising in WT vs. Npmc mice. To identify Myc dysregulated genes, we will use scRNA- seq to determine the transcriptional profiles of AMLs (obtained in Aim 1) initiated by WT vs. mutant Myc over- expression, either alone or in combination with Npmc. We will also perform ChIP-seq studies to characterize the genomic binding sites of WT vs. mutant Myc, and perform an integrated analysis of these datasets to define the relative contributions of direct vs. indirect effects of Myc on transcriptional outputs in AML cells. By combining the results of the functional studies (Aim 1) with the expression and DNA binding studies (Aim 2), we hope to better understand how Myc mutations alter transcription in hematopoietic cells, and how Npmc affects this output. If these studies are successful, they may provide important insights on MYC-mediated transformation, refine our current predictive tools for AML risk stratification, identify pathways involved in AraC sensitivity, and allow us to create new approaches for the treatment of AML patients.

项目摘要 该提案概述了一个5年的研究和职业发展计划，旨在支持候选人的 走向独立的学术生涯。拟议的研究项目，重点是急性 骨髓性白血病（AML）相关的MYC突变及其在AML细胞中的关键靶基因，将利用 关于华盛顿大学医学院的关键专业知识和可用资源。职业 发展计划和教学工作将为候选人提供各种技能，使其能够成功地 过渡到独立。这项提议是基于我们最近的观察，即一组独特的重复出现的 MYC错义突变在正常核型AML患者的优势克隆中显著富集 在我们的研究中，6/6的MYC突变与化疗相关， 发生NPM 1突变（NPMc），表明一种独特的协同性形式。根据我们初步的 研究发现，我们假设：1）过表达WT或突变MYC蛋白的AML在生物学上是不同的; 2） 每个MYC突变可能对其转录靶点有独特的影响; 3）MYC突变可能与其转录靶点协同作用。 NPMc突变。为了解决这些假设，我们提出以下具体目标： 具体目标1：我们将单独或联合研究AML相关MYC突变的功能。 关于NPMC我们已经开发了一个强力霉素诱导系统，使我们能够调节Myc的表达， 造血干细胞和祖细胞（HSPC），其含有或不含有Npmc突变。使用此 系统，我们将评估WT或突变Myc，有或没有Npmc，如何影响AML的发展，进展， 和AraC反应性。我们也在发展一种更符合生理的，有条件的 MYCT 58 N（AML和B细胞恶性肿瘤中的复发性突变）的基因敲入模型将使我们能够 探讨生理剂量的突变Myc和Npmc对AML发病机制和AraC敏感性的影响。 具体目标2：我们将定义WT与突变体的转录输出和基因组结合位点。 WT与Npmc小鼠中AML中的Myc蛋白。为了鉴定Myc失调基因，我们将使用scRNA- seq以确定由WT相对于突变Myc过表达启动的AML（在Aim 1中获得）的转录谱。 表达，单独或与Npmc组合。我们还将进行ChIP-seq研究，以表征 WT与突变Myc的基因组结合位点，并对这些数据集进行综合分析，以确定 Myc对AML细胞中转录输出的直接与间接作用的相对贡献。通过组合 功能研究（目的1）的结果与表达和DNA结合研究（目的2），我们希望 更好地了解Myc突变如何改变造血细胞的转录，以及Npmc如何影响这种输出。 如果这些研究是成功的，他们可能会提供重要的见解MYC介导的转化，完善我们的研究。 目前AML风险分层的预测工具，确定AraC敏感性相关的途径，并使我们能够 为AML患者的治疗创造新的方法。