Profiling of the chromatin landscape of human natural killer (NK) cells overexpressing the transcription factors T-BET or EOMES

T-BET and EOMES are key transcription factors in the development of mature NK cells in mice. However, the role of these transcription factors during human NK cell development is less well understood. Therefore, we overexpressed T-BET or EOMES in human umbilical cord blood-derived hematopoietic progenitor cells (HPC) and cultured them in vitro in an NK cell differentiation model. On day 21 of culture mature stage 4 (CD56+CD94+CD16-) and stage 5 (CD56+CD94+CD16+) NK cells from T-BET or EOMES overexpression and control cultures were sorted, whereafter genomic DNA was isolated and the chromatin accessibility landscape was determined by assay for transposase-accessible chromatin (ATAC) sequencing. Profiling of the epigenetic changes during T-BET or EOMES overexpression in mature NK cells revealed new insights in the regulatory role of T-BET and EOMES during terminal NK cell maturation. Profiling of the chromatin accessibility landscape by Fast-ATAC sequencing of stage 4 and stage 5 NK cells with T-BET or EOMES overexpression in comparison to control NK cells on day 21 of culture.

T-BET与EOMES是小鼠成熟自然杀伤（NK）细胞发育过程中的核心转录因子。然而，这两类转录因子在人类NK细胞发育中的调控作用尚未得到充分阐释。为此，我们在人脐带血来源的造血祖细胞（hematopoietic progenitor cells, HPC）中过表达T-BET或EOMES，并借助体外NK细胞分化模型进行培养。在培养至第21天时，我们分选获得过表达T-BET、过表达EOMES以及对照组的成熟第4型（CD56+CD94+CD16-）与第5型（CD56+CD94+CD16+）NK细胞；随后提取基因组DNA，通过转座酶可及性染色质测序（assay for transposase-accessible chromatin, ATAC）解析染色质可及性图谱。针对成熟NK细胞中T-BET或EOMES过表达后发生的表观遗传变化进行分析，为阐明二者在NK细胞终末成熟过程中的调控功能提供了全新视角。本研究通过Fast-ATAC测序，对比分析了培养第21天时，过表达T-BET、过表达EOMES的第4型与第5型NK细胞，以及对照组NK细胞的染色质可及性图谱。