Polito Maria Pia, Marini Grazia, Fabrizi Alessandra, Sercia Laura, Enzo Elena, De Luca Michele
Centre for Regenerative Medicine "Stefano Ferrari", Department of Life Science, University of Modena and Reggio Emilia, Modena, Italy.
Cell Death Dis. 2024 Jul 17;15(7):508. doi: 10.1038/s41419-024-06905-1.
Epidermal stem cells orchestrate epidermal renewal and timely wound repair through a tight regulation of self-renewal, proliferation, and differentiation. In culture, human epidermal stem cells generate a clonal type referred to as holoclone, which give rise to transient amplifying progenitors (meroclone and paraclone-forming cells) eventually generating terminally differentiated cells. Leveraging single-cell transcriptomic data, we explored the FOXM1-dependent biochemical signals controlling self-renewal and differentiation in epidermal stem cells aimed at improving regenerative medicine applications. We report that the expression of H1 linker histone subtypes decrease during serial cultivation. At clonal level we observed that H1B is the most expressed isoform, particularly in epidermal stem cells, as compared to transient amplifying progenitors. Indeed, its expression decreases in primary epithelial culture where stem cells are exhausted due to FOXM1 downregulation. Conversely, H1B expression increases when the stem cells compartment is sustained by enforced FOXM1 expression, both in primary epithelial cultures derived from healthy donors and JEB patient. Moreover, we demonstrated that FOXM1 binds the promotorial region of H1B, hence regulates its expression. We also show that H1B is bound to the promotorial region of differentiation-related genes and negatively regulates their expression in epidermal stem cells. We propose a novel mechanism wherein the H1B acts downstream of FOXM1, contributing to the fine interplay between self-renewal and differentiation in human epidermal stem cells. These findings further define the networks that sustain self-renewal along the previously identified YAP-FOXM1 axis.
表皮干细胞通过对自我更新、增殖和分化的严格调控来协调表皮更新和及时的伤口修复。在培养过程中,人类表皮干细胞产生一种称为全克隆的克隆类型,它产生短暂扩增祖细胞(形成亚克隆和旁克隆的细胞),最终产生终末分化细胞。利用单细胞转录组数据,我们探索了控制表皮干细胞自我更新和分化的FOXM1依赖性生化信号,旨在改善再生医学应用。我们报告说,在连续培养过程中,H1连接组蛋白亚型的表达会降低。在克隆水平上,我们观察到H1B是表达最多的亚型,特别是在表皮干细胞中,与短暂扩增祖细胞相比。事实上,在由于FOXM1下调而干细胞耗尽的原代表皮培养物中,其表达降低。相反,当通过强制表达FOXM1来维持干细胞区室时,H1B表达增加,无论是在来自健康供体的原代表皮培养物还是交界性大疱性表皮松解症(JEB)患者的培养物中。此外,我们证明FOXM1结合H1B的启动子区域,从而调节其表达。我们还表明,H1B与分化相关基因的启动子区域结合,并在表皮干细胞中负调节它们的表达。我们提出了一种新机制,其中H1B在FOXM1下游起作用,有助于人类表皮干细胞自我更新和分化之间的精细相互作用。这些发现进一步定义了沿着先前确定的YAP-FOXM1轴维持自我更新的网络。