Division of Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
Hoxworth Blood Center, University of Cincinnati College of Medicine, Cincinnati, Ohio, USA.
Stem Cells Transl Med. 2019 Jun;8(6):557-567. doi: 10.1002/sctm.18-0255. Epub 2019 Feb 21.
Bacterial and fungal infections are a major cause of morbidity and mortality in neutropenic patients. Donor-derived neutrophil transfusions have been used for prophylaxis or treatment for infection in neutropenic patients. However, the short half-life and the limited availability of large numbers of donor-derived neutrophils for transfusion remain a significant hurdle in the implementation of neutrophil transfusion therapy. Here, we investigate the in vitro and in vivo activity of neutrophils generated from human induced pluripotent stem cells (iPSC), a potentially unlimited resource to produce neutrophils for transfusion. Phenotypic analysis of iPSC-derived neutrophils reveal reactive oxygen species production at similar or slightly higher than normal peripheral blood neutrophils, but have an ∼50%-70% reduced Escherichia coli phagocytosis and phorbol 12-myristate 13-acetate induced formation of neutrophil extracellular traps (NET). Signaling of granulocytic precursors identified impaired AKT activation, but not ERK or STAT3, in agonist-stimulated iPSC-derived neutrophils. Expression of a constitutively activated AKT in iPSC-derived neutrophils restores most phagocytic activity and NET formation. In a model of bacterial induced peritonitis in immunodeficient mice, iPSC-derived neutrophils, with or without corrected AKT activation, migrate similarly to the peritoneal fluid as peripheral blood neutrophils, whereas the expression of activated AKT significantly improves their phagocytic activity in vivo. Stem Cells Translational Medicine 2019;8:557-567.
细菌和真菌感染是中性粒细胞减少症患者发病率和死亡率的主要原因。供体来源的中性粒细胞输注已被用于预防或治疗中性粒细胞减少症患者的感染。然而，半衰期短和大量供体来源的中性粒细胞用于输注的有限可用性仍然是中性粒细胞输注治疗实施的一个重大障碍。在这里，我们研究了来自人诱导多能干细胞（iPSC）的中性粒细胞的体外和体内活性，iPSC 是一种潜在的无限资源，可以产生用于输注的中性粒细胞。iPSC 衍生中性粒细胞的表型分析表明，活性氧物质的产生与正常外周血中性粒细胞相似或略高，但对大肠杆菌的吞噬作用降低了约 50%-70%，佛波醇 12-肉豆蔻酸 13-乙酸诱导的中性粒细胞胞外陷阱（NET）形成减少。在激动剂刺激的 iPSC 衍生中性粒细胞中，粒细胞前体的信号转导显示 AKT 激活受损，但 ERK 或 STAT3 不受影响。在 iPSC 衍生的中性粒细胞中表达组成型激活的 AKT 可恢复大多数吞噬活性和 NET 形成。在免疫缺陷小鼠细菌诱导性腹膜炎模型中，iPSC 衍生的中性粒细胞，无论是否纠正 AKT 激活，都像外周血中性粒细胞一样迁移到腹腔液中，而激活的 AKT 的表达显著改善了它们在体内的吞噬活性。《干细胞转化医学》2019；8:557-567.
