Division of Molecular Medicine and Gene Therapy, Lund Stem Cell Center, Lund University, Lund, Sweden.
Department of Research and Development, Central Blood Institute, Japanese Red Cross Society, Tokyo, Japan.
Commun Biol. 2021 Jun 3;4(1):677. doi: 10.1038/s42003-021-02202-1.
Immortalized erythroid cell lines are expected to be a promising source of ex vivo manufactured red blood cells (RBCs), however the induction of enucleation in these cell lines is inefficient at present. We utilized an imaging-based high-throughput system to identify chemical compounds that trigger enucleation of human erythroid cell lines. Among >3,300 compounds, we identified multiple histone deacetylase inhibitors (HDACi) inducing enucleated cells from the cell line, although an increase in membrane fragility of enucleated cells was observed. Gene expression profiling revealed that HDACi treatment increased the expression of cytoskeletal genes, while an erythroid-specific cell membrane protein, SPTA1, was significantly down-regulated. Restoration of SPTA1 expression using CRISPR-activation partially rescued the fragility of cells and thereby improved the enucleation efficiency. Our observations provide a potential solution for the generation of mature cells from erythroid cell lines, contributing to the future realization of the use of immortalized cell lines for transfusion therapies.
永生化红细胞系有望成为体外生产红细胞（RBC）的有前途的来源，但目前这些细胞系的去核效率不高。我们利用基于成像的高通量系统来鉴定可诱导人红细胞系去核的化合物。在 >3300 种化合物中，我们鉴定出多种组蛋白去乙酰化酶抑制剂（HDACi）可诱导细胞系中的去核细胞，但观察到去核细胞的膜脆性增加。基因表达谱分析显示，HDACi 处理增加了细胞骨架基因的表达，而红细胞特异性细胞膜蛋白 SPTA1 的表达则显著下调。使用 CRISPR 激活物恢复 SPTA1 的表达部分挽救了细胞的脆性，从而提高了去核效率。我们的观察结果为从红细胞系生成成熟细胞提供了一种潜在的解决方案，有助于实现未来使用永生化细胞系进行输血治疗。
