Cancer evolution is a multifaceted process leading to dysregulation of cellular expansion and differentiation through somatic mutations and epigenetic dysfunction. Clonal expansion and evolution is driven by cell-intrinsic and -extrinsic selective pressures, which can be captured with increasing resolution by single-cell and bulk DNA sequencing. Despite the extensive genomic alterations revealed in profiling studies, there remain limited experimental systems to model and perturb evolutionary processes. Here, we integrate multi-recombinase tools for reversible, sequential mutagenesis from premalignancy to leukemia. We demonstrate that inducible Flt3 mutations differentially cooperate with Dnmt3a, Idh2, and Npm1 mutant alleles, and that changing the order of mutations influences cellular and transcriptional landscapes. We next use a generalizable, reversible approach to demonstrate that mutation reversion results in rapid leukemic regression with distinct differentiation patterns depending upon co-occurring mutations. These studies provide a path to experimentally model sequential mutagenesis, investigate mechanisms of transformation and probe oncogenic dependency in disease evolution.

中文翻译：

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造血系统恶性肿瘤中亚克隆肿瘤发生和依赖性的体内模型


癌症进化是一个多方面的过程，通过体细胞突变和表观遗传功能障碍导致细胞扩增和分化失调。克隆扩增和进化由细胞内源性和外源性选择压力驱动，可以通过单细胞和大量 DNA 测序以更高的分辨率捕获。尽管在分析研究中揭示了广泛的基因组改变，但用于建模和扰乱进化过程的实验系统仍然有限。在这里，我们整合了多重组酶工具，用于从癌前病变到白血病的可逆、序贯诱变。我们证明，诱导型 Flt3 突变与 Dnmt3a、Idh2 和 Npm1 突变等位基因不同合作，并且改变突变顺序会影响细胞和转录景观。接下来，我们使用一种可推广的、可逆的方法来证明突变逆转导致快速白血病消退，根据同时发生的突变，具有不同的分化模式。这些研究为实验模拟顺序诱变、研究转化机制和探测疾病进化中的致癌依赖性提供了一条途径。