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，从而导致随后的克隆性扩张。是这样的 随着患者年龄的增长，外周血样本中很容易观察到克隆性扩张。这 这种现象被称为克隆性造血(CH)，会增加个体患血液病的风险 恶性肿瘤和心血管疾病，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)中更频繁发生突变的基因 白血病(AML)是与DNA甲基化相关的基因，如DNMT3A。然而，这一过程通过 目前尚不清楚是哪种克隆性造血作用发生并导致了这些疾病。这个项目试图揭开 这些体细胞突变对允许血细胞转化的造血的生物学影响 转化为恶性肿瘤并影响心血管健康。具体地说，这个项目的目的是描述 DNMT3A突变对基因转录、DNA甲基化和染色质可及性的影响 从克隆性造血到疾病的进展。 该方案的主要方法是单细胞测序和个体基因分型。 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本基因分型方法来比较 野生型和突变型造血祖细胞的转录图谱。这将为我们提供对 分化命运扭曲，以及与体细胞突变相关的基因表达的变化。在……里面 具体目标2，我将使用我们实验室开发的类似方法来结合单细胞甲基组和染色质 使用体细胞基因分型的可及性分析，以确定可以解释 转录和细胞命运决定的变化。这项研究的结果将告诉我们致病机制 从正常造血到CH再到恶性肿瘤，为筛查CH以及 可能的治疗选择，以防止疾病的发展。这个项目将是理想的有抱负的 医生-科学家正在接受培训，因为它融合了应用于临床样本的新测序技术 使用尖端的计算生物学方法进行分析，及其可能对临床的影响 建议。在我的赞助人、共同赞助人、论文委员会、Tri-I MD的领导下- 博士课程，以及这次奖学金的支持，我相信我会做好充分的准备，继续并实现我的 目标是成为一名内科医生、科学家和独立研究者。

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.