Project 3: Contribution of inflammation and DNA damaging factors to clonal expansion and malignant transformation in a community cohort of older adults

项目摘要/摘要 项目3将检验这一假设，即可改变的风险因素，包括炎症和DNA损伤因素， 某些基因突变对造血干/祖细胞克隆扩增的影响 它们会转化为血液系统的恶性肿瘤。我们将确定与HSPC相关的可修改的风险因素 对9,513人(Black)进行的前瞻性流行病学研究中的克隆性扩张和恶变 27%，女性55%)现在都是75岁，来自ARIC，这是一项长期的社区队列研究。我们将使用存储的 用于单细胞DNA测序的棕黄色涂层，并利用完整的外周血白细胞(PBL)整体 外显子组测序(WES)、甲基化和蛋白质组谱。目标1将决定克隆的长期变化 单细胞水平的成分和来自Core A的资源，并识别炎症和DNA损伤- 与这些变化相关的相关因素。对于250名ARIC参与者，同时具有1个WES克隆变体 1或在时间2(20年后)和/或时间2之后的血液系统恶性病变时，我们将测量单个白细胞 用单细胞DNA测序法测定两个时间点的变异图谱。通过以前所未有的速度跟踪单个克隆 解决方案中，我们将评估克隆适合性，并确定可修改的风险因素，用于克隆的扩展 变种。我们的研究是第一次研究血液系统恶性肿瘤患者克隆性成分的变化。 社区队列。在目标2中，我们将评估克隆组成、变异等位基因频率之间的关系 (VAFs)，以及已知的髓系恶性肿瘤驱动基因和新的血液病复发变异的变化 在二十年的时间里有恶性的风险。在9,513名参与者中，我们将在时间1和 2(相隔20年)，以确定具有变化的VAFs的新的复发变异，并根据克隆对参与者进行分类 已知司机基因的组成和VAFs。对于驱动基因，我们将估计克隆性变化的关联 随着时间的推移，成分和VAF与血液系统恶性肿瘤的风险有关。我们预计341个第一小学和66个第二小学 在我们的队列中有恶性血液病。AIM 3将评估炎症和DNA之间的联系 携带可检测克隆变异者的血液系统恶性肿瘤风险的损害相关因素。到了时间2， 我们预计，10%的参与者将在DIVER和新基因中具有WES可以检测到的克隆变异。我们会 评估携带和不携带特定基因的人群中可修饰因子与血液系统恶性肿瘤的相关性 克隆变种超过30年。项目3将确定需要测试的药物作为克隆扩增和 项目1和2中的小鼠恶性转化。我们还将测试改变的细胞因子之间的关系 和项目1中确定的CpG甲基化，并确定是否观察到项目2中确定的突变 暴露在DNA损伤环境中的参与者。该项目将广泛利用核心A进行单细胞分析 核心B用于协调。确定驱动克隆性扩张和恶性的可修改因素 转化将提供拦截克隆扩张和恶性转化的机会，并使 早期发现风险较高的人，因为他们的遗传和可修改的风险因素概况。

Project Summary/Abstract Project 3 will test the hypothesis that modifiable risk factors, including inflammation and DNA-damaging factors, influence expansion of hematopoietic stem and progenitor cell (HSPC) clones with certain genetic mutations and their transformation to hematologic malignancy. We will identify modifiable risk factors associated with HSPC clonal expansion and malignant transformation in a prospective epidemiologic study of 9,513 persons (Black 27%, female 55%) now all >75 years old from ARIC, a long-standing community cohort study. We will use stored buffy coat for single cell DNA sequencing, and leverage completed peripheral blood leukocyte (PBL) whole exome sequencing (WES), methylation, and proteomic profiling. Aim 1 will determine long-term changes in clonal composition at the single-cell level with resources from Core A and identify inflammation and DNA damage- related factors associated with those changes. For 250 ARIC participants with ³1 clonal variant by WES at Time 1 or at Time 2 (20 years later) and/or hematologic malignancy after Time 2, we will measure individual leukocyte variant profiles at both times by single cell-DNA sequencing. By tracking individual clones at unprecedented resolution, we will assess clonal fitness and identify modifiable risk factors for expansion of clones with specific variants. Our study is the first to address change in clonal composition with hematologic malignancy in a community cohort. In Aim 2, we will evaluate the association of clonal composition, variant allele frequencies (VAFs), and changes of known myeloid malignancy driver genes and novel recurrent variants with hematologic malignancy risk over the span of two decades. In 9,513 participants, we will interrogate WES data at Time 1 and 2 (20 years apart) to identify novel recurrent variants with changing VAFs and classify participants by clonal composition and VAFs of known driver genes. For driver genes, we will estimate associations of change in clonal composition and VAF over time with hematologic malignancy risk. We project 341 first and 66 second primary hematologic malignancies within our cohort. Aim 3 will evaluate the association of inflammation- and DNA damage-related factors with hematologic malignancy risk in persons with detectable clonal variants. By Time 2, we expect >10% of participants will have clonal variants detectable by WES in driver and novel genes. We will estimate the association of modifiable factors with hematologic malignancy in persons with and without specific clonal variants over 30 years. Project 3 will identify drugs to test as interventions for clonal expansion and malignant transformation in mice in Projects 1 and 2. We also will test associations between altered cytokines and CpG methylation identified in Project 1 and determine whether mutations identified in Project 2 are observed in participants with DNA-damaging exposures. This Project will extensively utilize Core A for single-cell analysis and Core B for coordination. Identifying modifiable factors that drive clonal expansion and malignant transformation will provide opportunities to intercept clonal expansion and malignant transformation and enable early detection in persons at higher risk due to their genetic and modifiable risk factor profile.