Selective pressures from inherited variation impacting myeloproliferative neoplasm initiation

项目总结/摘要 虽然在描述驱动骨髓增生的体细胞突变方面取得了实质性进展， 流行病学研究表明，MPN是一种重要的遗传性疾病。我们 最近进行了最大的MPN全基因组关联研究，以确定遗传风险变异 促进这种疾病的发生。我们的初步分析表明，HSC自我调节的作用， 通过这些遗传的风险变体来更新和发挥作用。然而，我们缺乏一个完整的机械理解 这些遗传变异产生的选择压力如何促进MPN的获得。这里在 在R 01中，我们特别想了解遗传风险变体如何提供选择性压力， MPN启动。在这项资助的第一个目标中，我们试图使用严格的小鼠模型来定义一个变体- 含有HSC选择性增强子的Gfi 1b可以促进MPN的获得。我们将研究这种增强剂 在天然造血中起作用，以促进MPN启动，以及这种增强子如何与 基因突变在这个过程中在这项资助的第二个目标中，我们试图了解功能丧失是如何 CHEK 2的基因组编辑可以通过偶联原代人的基因组编辑来促进MPN获得和克隆造血。 造血干细胞/祖细胞和全基因组测序，以评估这些生殖系遗传 突变可以促进肿瘤发生。最后，在第三个目标中，我们寻求更全面地定义变异- 携带调控元件和靶基因的HSC可能通过转录因子在HSC自我更新和功能中发挥作用。 多重向导交换Cas9基因组编辑方法在原代人造血干细胞中的用途， 祖细胞总的来说，我们在这项资助中提出的研究将为遗传风险如何提供新的见解。 变体可以提供使MPN能够出现的选择压力。

Project Summary/ Abstract While substantial progress has been made to delineate the somatic mutations that drive myeloproliferative neoplasms (MPNs), epidemiologic studies demonstrate a significant heritable risk for disease acquisition. We have recently conducted the largest genome-wide association study of MPNs to define inherited risk variants that promote the acquisition of this disease. Our initial analyses suggest a role for modulation of HSC self- renewal and function by these inherited risk variants. However, we lack a complete mechanistic understanding of how the selective pressures that arise from these inherited variants can promote MPN acquisition. Here, in this R01, we specifically want to understand how inherited risk variants can provide selective pressures to enable MPN initiation. In the first aim of this grant, we seek to use rigorous mouse models to define how a variant- containing HSC-selective enhancer of Gfi1b can promote MPN acquisition. We will study how this enhancer plays a role in native hematopoiesis to promote MPN initiation, as well as how this enhancer can cooperate with somatic mutations in this process. In the second aim of this grant, we seek to understand how loss-of-function of CHEK2 can promote MPN acquisition and clonal hematopoiesis by coupling genome editing of primary human hematopoietic stem/ progenitor cells and whole genome sequencing to assess how such germline genetic mutations can promote oncogenesis. Finally, in the third aim, we seek to more holistically define how variant- harboring regulatory elements and target genes may play a role in HSC self-renewal and function through the use of a multiplexed guide-swap Cas9 genome editing approach in primary human hematopoietic stem and progenitor cells. Collectively, the studies we propose in this grant will provide new insights into how inherited risk variants can provide selection pressures that enable MPNs to arise.