Functional effects of DNMT3A R882 on human haematopoietic stem and progenitor cells myeloid differentiation

This data presents the transcriptomes of bulk mature colonies derived from single human haematopoietic stem cells / multipotent progenitors (HSC/MPPs) and granulocyte macrophage progenitors (GMPs) purified from one indiviual with age related clonal haematopoisesis. The profiled colonies contain mature monocytes, as measured by conventional cell surface markers (CD14+, CD15-, GlyA-), but no other mature blood cell types. This dataset is part of a larger study, the main objective of which is to understand the functional effects conferred by somatic DNMT3A R882H mutation on human haematopoietic stem cell differentiation capacity. In clonal haematopoiesis, DNMT3A R882H and DNMT3A WT HSPCs co-exist in the same individual. We compared the transcriptional differences between mature monocytic colonies derived from DNMT3A R882H and WT HSPCs in vitro. Analysis of this dataset shows that, in this individual, DNMT3A R882H HSPCs produce less mature monocytes than their WT counterparts over the same culture time. 39 bulk monocyte colonies derived from 33 single HSC/MPPs and 6 single GMPs were analysed from 1 individual. Single HSC/MPPs (CD19-, CD33-, CD34+, CD45dim, CD38-, CD45RA+), and GMPs (CD19-, CD33-, CD34+, CD45dim, CD38+, CD7/CD10-, CD45RA+) were cultured in vitro for 21 days in myelo-erythroid medium (MEM), colonies containing mature monocytes but no other mature blood cell lineage were selected and harvested into RNA extraction plates and RNA extracted using the Qiagen RNeasy protocol.

本数据集呈现了从1名年龄相关性克隆性造血（clonal haematopoiesis）个体体内纯化得到的单个人类造血干细胞/多能祖细胞（HSC/MPPs）以及粒细胞-巨噬细胞祖细胞（GMPs）所衍生的批量成熟集落的转录组。经经典细胞表面标志物（CD14+、CD15-、GlyA-）鉴定，所分析的集落仅包含成熟单核细胞，不含有其他成熟血细胞类型。本数据集隶属于一项更大规模的研究，该研究的核心目标是阐明体细胞DNMT3A R882H突变对人类造血干细胞分化潜能所赋予的功能影响。在克隆性造血过程中，DNMT3A R882H突变型与DNMT3A野生型（WT）造血干祖细胞（HSPCs）可共存于同一患者体内。本研究对体外培养的、分别源自DNMT3A R882H突变型与WT HSPCs的成熟单核细胞集落之间的转录差异进行了比较。对本数据集的分析结果表明，在该个体中，相同培养时长下，DNMT3A R882H HSPCs所产生的成熟单核细胞数量少于其野生型对应群体。本研究对来自1名个体的33个单细胞来源HSC/MPPs与6个单细胞来源GMPs所衍生的39个批量单核细胞集落进行了分析。单克隆HSC/MPPs（标志物为CD19-、CD33-、CD34+、CD45dim、CD38-、CD45RA+）与GMPs（标志物为CD19-、CD33-、CD34+、CD45dim、CD38+、CD7/CD10-、CD45RA+）在髓系-红系培养基（MEM）中体外培养21天；随后筛选出仅含成熟单核细胞、无其他成熟血细胞谱系的集落，将其收集至RNA提取板中，并采用Qiagen RNeasy试剂盒操作流程提取RNA。