利用突变的慢性髓性白血病源性诱导多能干细胞（iPSCs）建立急变期白血病模型。

Modeling Blast Crisis Using Mutagenized Chronic Myeloid Leukemia-Derived Induced Pluripotent Stem Cells (iPSCs).

机构信息

INSERM UMR-S-1310, Université Paris Saclay, 94800 Villejuif, France and ESTeam Paris Sud, Université Paris Saclay, 94800 Villejuif, France.

APHP Paris Saclay, Department of Hematology, Hôpital Bicêtre & Paul Brousse, 94800 Villejuif, France.

出版信息

Cells. 2023 Feb 12;12(4):598. doi: 10.3390/cells12040598.

PURPOSE

To model CML progression in vitro and generate a blast crisis (BC-CML) model in vitro in order to identify new targets.

METHODS

Three different CML-derived iPSC lines were mutagenized with the alkylating agent ENU on a daily basis for 60 days. Cells were analyzed at D12 of hematopoietic differentiation for their phenotype, clonogenicity, and transcriptomic profile. Single-cell RNA-Seq analysis has been performed at three different time points during hematopoietic differentiation in ENU-treated and untreated cells.

RESULTS

One of the CML-iPSCs, compared to its non-mutagenized counterpart, generated myeloid blasts after hematopoietic differentiation, exhibiting monoblastic patterns and expression of cMPO, CD45, CD34, CD33, and CD13. Single-cell transcriptomics revealed a delay of differentiation in the mutated condition as compared to the control with increased levels of (mesodermal marker) and a decrease in and . Bulk transcriptomics analyzed along with the GSE4170 GEO dataset reveal a significant overlap between ENU-treated cells and primary BC cells. Among overexpressed genes, was identified, and its relevance was confirmed in a cohort of CML patients.

CONCLUSIONS

iPSCs are a valuable tool to model CML progression and to identify new targets. Here, we show the relevance of CD25 identified in the iPSC model as a marker of CML progression.

目的

建立体外 CML 进展模型并生成体外急变期 CML（BC-CML）模型，以鉴定新的靶点。

方法

将三种不同的 CML 来源的 iPSC 线用烷化剂 ENU 进行每日诱变，持续 60 天。在造血分化的第 12 天分析细胞的表型、集落形成能力和转录组谱。在造血分化的三个不同时间点对 ENU 处理和未处理的细胞进行单细胞 RNA-Seq 分析。

结果

与未突变的对照相比，一种 CML-iPSC 在造血分化后产生了髓性白血病细胞，表现出单核细胞样形态和 cMPO、CD45、CD34、CD33 和 CD13 的表达。单细胞转录组学显示突变条件下的分化延迟，与对照相比，（中胚层标志物）水平升高，和水平降低。与 GSE4170 GEO 数据集一起分析的批量转录组学揭示了 ENU 处理的细胞与原发性 BC 细胞之间存在显著重叠。在过表达的基因中，鉴定到，并在一组 CML 患者中证实了其相关性。