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.

项目摘要/摘要 人类癌症是通过连续获得体细胞突变而发展起来的，这些突变产生了克隆细胞 通过克隆选择和进化的反复过程，生长超过正常细胞的种群。而当 长期以来，人们已经认识到肿瘤发生的多步骤性质，这是支撑这一过程的基本原则 对这一进程及其结果的确定仍不完全清楚。现在很明显，从大规模的 突变共生的癌症基因组测序及突变获取的相对时机 遵循非随机模式，但有限的研究涉及突变顺序和合作的影响 在生物学或临床结果上。 白血病发生学提供了一个有吸引力的案例来模拟和研究克隆进化的主要原则。 白血病的基因组非常简单，与固体相比，携带的司机遗传损害的数量非常少。 肿瘤。更具体地说，急性髓系白血病(AML)基因组可以含有少至2-3个驱动基因 损伤。我的实验室最近开发了一种使用序列CRISPR介导的AML克隆进化模型 人类ipscs的基因编辑。我们发现，通过将3个驱动程序突变引入正常的IPSC(ASXL1 C- 终结点截断，SRSF2P95L和NRASG12DAMLSAR)，我们可以创建一个“从头”可刻。此外，我们的 初步数据表明，特定突变之间的合作和/或它们获得的顺序 对白血病的发生施加限制。 这项建议的首要目标是研究突变获得顺序在 急性髓系白血病的克隆性进化。在目标1中，我们将检验这样的假设，即启动突变建立一个 后来的突变(S)需要在其上作用以建立我们的IPSC中的白血病状态的表观遗传格局 模型和原代人造血干/祖细胞(HSPC)。目标2将定义最低要求 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的获取顺序来激活突变 和NRAS突变在我们的新的肿瘤发生模型中。 这项工作利用了我实验室开发的一种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。