Deciphering Clonal Hematopoiesis via Single-Cell Multi-omics

项目摘要 几乎所有的组织都会获得自发的体细胞突变，导致随后的克隆扩增。等 随着患者年龄的增加，在外周血样品中容易观察到克隆扩增。这 这种现象，称为克隆造血（CH），增加了个体的血液系统疾病的风险。 恶性肿瘤和心血管疾病，并且在65岁以上的患者中患病率超过10%。 在CH以及随后的恶性肿瘤如急性髓系白血病中， 白血病（AML）是与DNA甲基化相关的基因，如DNMT3A。然而，通过 哪种克隆造血发生并引起这些疾病是未知的。该项目旨在揭示 这些体细胞突变对造血的生物学影响，使血液细胞转化为 恶性肿瘤并影响心血管健康。具体而言，该项目旨在描述 DNMT3A突变对基因转录、DNA甲基化和染色质可及性的影响， 从克隆造血到疾病的发展。 该建议的主要方法围绕单细胞测序，沿着个体的基因分型。 细胞在具体目标1中，我将使用我们实验室最近开发的转录组基因分型方法来比较 野生型和突变型造血祖细胞的转录景观。这将提供深入了解 分化命运偏斜以及与体细胞突变相关的基因表达的变化。在 具体目标2，我将使用我们实验室开发的类似方法来联合收割机结合单细胞甲基化组和染色质 可及性分析与体细胞基因分型，以确定推定的机制，可以解释 转录和细胞命运决定的变化。这项研究的结果将告知发病机制， 正常造血到CH到恶性肿瘤，为CH筛查的作用以及 可能的治疗选择，以防止疾病的进展。这个项目将是理想的一个有抱负的 由于其混合了应用于临床样本的新型测序技术， 使用尖端的计算生物学方法进行分析，沿着， 建议.在我的赞助商、共同赞助商、论文委员会、三I医学博士领导的指导下， 博士课程，和这个奖学金的支持，我相信我将做好充分的准备，追求和实现我的 目标是成为一名医生科学家和独立调查员。

Project Summary Almost all tissues acquire spontaneous somatic mutations that can result in subsequent clonal expansion. Such clonal expansions are readily observable in peripheral blood samples as a patient’s age increases. This phenomenon, referred to as clonal hematopoiesis (CH), increases an individual’s risk for hematologic malignancies and cardiovascular disease, and has a prevalence over 10% in patients over the age of 65. Amongst the more frequently mutated genes in CH, as well as in subsequent malignancies such as acute myeloid leukemia (AML), are genes associated with DNA methylation, such as DNMT3A. However, the process through which clonal hematopoiesis occurs and gives rise to these diseases is unknown. This project seeks to uncover the biological impacts of these somatic mutations on hematopoiesis, which allow hematologic cells to transform into malignancies and impact cardiovascular health. Specifically, this project aims to characterize the effects of DNMT3A mutations on gene transcription, DNA methylation, and chromatin accessibility, to provide insight into the progression from clonal hematopoiesis to disease. The main approach in this proposal centers around single-cell sequencing along with genotyping of individual cells. In Specific Aim 1, I will use our lab’s recently developed Genotyping of Transcriptomes method to compare the transcriptional landscape of wild-type and mutant hematopoietic progenitor cells. This will provide insight into differentiation fate skews as well as changes in gene expression associated with the somatic mutations. In Specific Aim 2, I will use similar methods developed in our lab to combine single-cell methylome and chromatin accessibility profiling with somatic genotyping in order to identify putative mechanisms that could explain the changes in transcription and cell-fate determination. The outcomes of this study will inform the pathogenesis of normal hematopoiesis to CH to malignancy, providing useful insights into the role of screening for CH as well as possible therapeutic options to prevent the progression of disease. This project will be ideal for an aspiring physician-scientist in training, given its blend of novel sequencing technologies applied to clinical samples analyzed using cutting-edge computational biological methods, along with its possible implications for clinical recommendations. With the mentorship of my sponsor, co-sponsor, thesis committee, leadership of the Tri-I MD- PhD program, and the support of this fellowship, I am confident I will be well prepared to pursue and achieve my goal of being a physician scientist and independent investigator.