Impact of mutational order on molecular mechanisms of oncogenesis

突变顺序对肿瘤发生分子机制的影响

• 批准号: 10375146
• 负责人: Eirini Papapetrou
• 金额: $ 66.53万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-10 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10375146
• 关键词: Acute Myelocytic Leukemia; Address; Affect; Alleles; Biological; Biological Assay; Cells; Chromatin; Clinical; Clonal Evolution; Clone Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Data; Epigenetic Process; Event; Evolution; Gene Expression; Genes; Genetic; Genome; Goals; Hematopoietic stem cells; Human; In Vitro; Laboratories; Large-Scale Sequencing; Lesion; Malignant Neoplasms; Mediating; Modeling; Molecular; Mutation; Myelogenous; Nature; Normal Cell; Outcome; Pattern; Population; Population Genetics; Process; Regulator Genes; Reporting; Role; SRSF2 gene; Sampling; Shapes; Signal Transduction; Solid; Solid Neoplasm; Somatic Mutation; Testing; Work; base; cancer genome; driver mutation; genetic analysis; in vivo; induced pluripotent stem cell; leukemia; leukemogenesis; multiple omics; mutant; single-cell RNA sequencing; transcriptome; tumorigenesis

PROJECT SUMMARY/ABSTRACT Human cancers develop through the successive acquisition of somatic mutations which give rise to clonal cell populations that outgrow normal cells through a reiterative process of clonal selection and evolution. While the multi-step nature of oncogenesis has been long recognized, the fundamental principles that underlie this process and determine its outcomes remain incompletely understood. It is now clear from large-scale sequencing of cancer genomes that mutational co-occurrence and the relative timing of mutational acquisition follow non-random patterns, but limited studies have addressed the impact of mutational order and cooperation in biological or clinical outcomes. Leukemogenesis offers an attractive case to model and study overarching principles of the clonal evolution. Leukemias have very simple genomes, carrying very low numbers of driver genetic lesions compared to solid tumors. More specifically, acute myeloid leukemia (AML) genomes can harbor as few as 2-3 driver genetic lesions. My lab recently developed a model of clonal evolution of AML using sequential CRISPR-mediated gene editing of human iPSCs. We found that by introducing 3 driver mutations into normal iPSCs (ASXL1 C- terminus truncation, SRSF2P95L and NRASG12D SAR) we can create a “de novo” engraftable AML. Furthermore, our preliminary data suggest that cooperation between specific mutations and/or the order of their acquisition impose constraints on leukemogenesis. The overarching goal of this proposal is to investigate the role of the order of mutational acquisition in the clonal evolution of AML. In Aim 1, we will test the hypothesis that the initiating mutation establishes an epigenetic landscape upon which the later mutation(s) need to act to establish a leukemic state in our iPSC model and in primary human hematopoietic stem and progenitor cells (HSPCs). Aim 2 will define the minimum number of mutations required for leukemogenesis. In Aim 3 we will explore the mechanistic constraints underlying “obligatory late” signaling activating mutations by varying the order of acquisition of ASXL1, SRSF2 and NRAS mutations in our de novo oncogenesis model. This work harnesses a unique myeloid leukemogenesis model developed in my laboratory to address fundamental questions on the effects of mutational order on leukemogenesis and oncogenesis in general.

项目概要/摘要 人类癌症是通过连续获得体细胞突变而产生的，从而产生克隆细胞 通过克隆选择和进化的重复过程，细胞群的生长速度超过正常细胞。虽然 肿瘤发生的多步骤性质早已被认识到，其基本原理 过程及其结果的确定仍不完全清楚。现在从大规模来看 对癌症基因组进行测序，确定突变共现和突变获得的相对时间 遵循非随机模式，但有限的研究已经解决了突变顺序和合作的影响 生物学或临床结果。 白血病发生为建模和​​研究克隆进化的总体原理提供了一个有吸引力的案例。 白血病具有非常简单的基因组，与实体瘤相比，携带的驱动基因病变数量非常少 肿瘤。更具体地说，急性髓系白血病 (AML) 基因组可包含少至 2-3 个驱动基因 病变。我的实验室最近开发了一种使用连续 CRISPR 介导的 AML 克隆进化模型 人类 iPSC 的基因编辑。我们发现，通过将 3 个驱动突变引入正常 iPSC（ASXL1 C- 末端截断（SRSF2P95L 和 NRASG12D SAR），我们可以创建“从头”可植入 AML。此外，我们的 初步数据表明，特定突变和/或其获得顺序之间的合作 对白血病发生施加限制。 该提案的总体目标是研究突变获得顺序在 AML 的克隆进化。在目标 1 中，我们将检验以下假设：起始突变建立了 后续突变需要发挥作用才能在 iPSC 中建立白血病状态的表观遗传景观 模型和原代人类造血干细胞和祖细胞（HSPC）中。目标 2 将定义最小值 白血病发生所需的突变数量。在目标 3 中，我们将探索机械约束 通过改变 ASXL1、SRSF2 的获取顺序，潜在的“强制性晚期”信号激活突变 以及我们的从头肿瘤发生模型中的 NRAS 突变。 这项工作利用我实验室开发的独特的髓系白血病发生模型来解决 关于突变顺序对白血病发生和肿瘤发生的影响的基本问题。