Gene expression profile at single cell level of Lin-Sca-1+c-Kit+ cells (LSKs) from miR-142 WT CML mice (CP LSC) and of LSKs from miR-142 KO CML mice (BC LSC). [scRNA-Seq]

To assess how the miR-142 deficit affected the BM LSC-enriched LSK landscape, we conducted a single cell RNA-seq analysis of BM miR-142+/+ BCR-ABL (CP CML) and miR-142-/- BCR-ABL (BC CML) LSKs which were harvested from the respective mice, 2 weeks after BCR-ABL induction. Overall design: ~8000 cells per sample were captured on a 10x Genomics Chromium using a Next GEM Single Cell 3' GEM kit V3.1 (10xGenomics). Using the Leiden clustering algorithm to identify groups of cells with similar transcriptomes, we identified 8 distinct LSK clusters. We annotated these clusters into distinct lineage-primed subsets using the expression levels of hematopoietic gene transcription factors and cluster differentiation (CD) antigens.

为评估miR-142缺陷对骨髓（Bone Marrow, BM）白血病干细胞（Leukemia Stem Cell, LSC）富集的LSK细胞图谱的影响，我们对BCR-ABL诱导2周后分别从对应小鼠体内获取的miR-142野生型（miR-142+/+）BCR-ABL阳性慢性髓系白血病慢性期（CP CML）与miR-142敲除型（miR-142-/-）BCR-ABL阳性慢性髓系白血病急变期（BC CML）的LSK细胞，开展了单细胞RNA测序分析。本研究整体实验设计如下：每个样本约8000个细胞通过10x Genomics Chromium平台，使用Next GEM Single Cell 3' GEM试剂盒V3.1（10xGenomics）完成单细胞捕获。通过Leiden聚类算法识别转录组特征相似的细胞群，最终鉴定出8个不同的LSK细胞簇。我们借助造血基因转录因子的表达水平以及簇分化抗原（Cluster of Differentiation, CD）的表达特征，将这些细胞簇注释为不同的谱系定向亚群。