Liu Xiaohui, Sha Jinghan, Wang Luxiang, Wang Zixuan, Fang Zhou, Han Xiao, Tan Shuiyi, Chen Yi, Yuan Hao, De The Hugues, Zhou Jun, Zhu Jun
Shanghai Institute of Hematology, State Key Laboratory of Medical Genomics, National Research Center for Translational Medicine at Shanghai, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China; CNRS IRP (International research Project), Cancer, Aging and Hematology, Sino-French Research Center for Life Sciences and Genomics, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025.
Shanghai Institute of Hematology, State Key Laboratory of Medical Genomics, National Research Center for Translational Medicine at Shanghai, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025.
Haematologica. 2025 Feb 1;110(2):385-396. doi: 10.3324/haematol.2024.285438.
The ubiquitination or SUMOylation of hematopoietic-related factors plays pivotal roles in hematopoiesis. RNF111, known as a ubiquitin ligase, is a newly discovered SUMO-targeted ubiquitin ligase involved in multiple signaling pathways mediated by transforming growth factor (TGF)-β family members. However, its role in hematopoiesis remains unclear. Herein, a heritable Rnf111 mutant zebrafish line was generated by CRISPR/Cas9-mediated genome editing. Impairment of hematopoietic stem and progenitor cells (HSPC) of definitive hematopoiesis was found in Rnf111-deficient mutants. Ablation of Rnf111 resulted in decreased phosphorylation of Smad2/3 in HSPC. Definitive endoderm 2 inducer (IDE2), which specifically activates TGF-β signaling and downstream Smad2 phosphorylation, could restore definitive hematopoiesis in Rnf111-deficient embryos. Further molecular mechanism studies revealed that Gcsfr/NO signaling was an important target pathway of Smad2/3 involved in Rnf111-mediated HSPC development. In conclusion, our study demonstrated that Rnf111 contributes to the development of HSPC by maintaining Smad2/3 phosphorylation and activation of the Gcsfr/NO signaling pathway.
造血相关因子的泛素化或类泛素化修饰在造血过程中起着关键作用。RNF111作为一种泛素连接酶，是新发现的一种靶向类泛素的泛素连接酶，参与由转化生长因子（TGF）-β家族成员介导的多种信号通路。然而，其在造血过程中的作用仍不清楚。在此，通过CRISPR/Cas9介导的基因组编辑构建了一种可遗传的Rnf111突变斑马鱼品系。在Rnf111缺陷型突变体中发现了定向造血的造血干细胞和祖细胞（HSPC）受损。Rnf111的缺失导致HSPC中Smad2/3磷酸化水平降低。特异性激活TGF-β信号及下游Smad2磷酸化的定向内胚层2诱导剂（IDE2），可恢复Rnf111缺陷型胚胎的定向造血。进一步的分子机制研究表明，Gcsfr/NO信号是Smad2/3参与Rnf111介导的HSPC发育的重要靶标信号通路。总之，我们的研究表明，Rnf111通过维持Smad2/3磷酸化以及激活Gcsfr/NO信号通路，促进HSPC的发育。
