Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
Biol Blood Marrow Transplant. 2013 Aug;19(8):1210-9. doi: 10.1016/j.bbmt.2013.05.012. Epub 2013 May 22.
Hepatic iron overload is common in patients undergoing hematopoietic cell transplantation. We showed previously in a murine model that transplantation of allogeneic T cells induced iron deposition and down-regulation of hepcidin (Hamp) in hepatocytes. We hypothesized that hepatic injury was related to disrupted iron homeostasis triggered by the interaction of Fas-ligand, expressed on activated T cells, with Fas on hepatocytes. In the current study, we determined the effects of modified expression of the Flice inhibitory protein (FLIP long [FLIPL]), which interferes with Fas signaling, on the impact of Fas-initiated signals on the expression of IL-6 and Stat3 and their downstream target, Hamp. To exclude a possible contribution by other pathways, we used agonistic anti-Fas antibodies (rather than allogeneic T cells) to trigger Fas signaling. Inhibition of FLIPL by RNA interference resulted, as expected, not only in enhanced hepatocyte apoptosis in response to Fas signals, but also in decreased levels of IL-6, Stat3, and Hamp. In contrast, overexpression of FLIPL protected hepatocytes against agonistic anti-Fas antibody-mediated apoptosis and increased the levels of IL-6 and Stat3, thereby maintaining the expression of Hamp in an NF-κB-dependent fashion. In vivo overexpression of FLIPL in the liver via hydrodynamic transfection, similarly, interfered with Fas-initiated apoptosis and prevented down-regulation of IL-6, Stat3, and Hamp. These data indicate that Fas-dependent signals alter the regulation of iron homeostasis and suggest that signals initiated by Fas may contribute to peritransplantation iron accumulation.
肝铁过载在接受造血细胞移植的患者中很常见。我们之前在小鼠模型中表明,同种异体 T 细胞的移植会导致铁沉积和肝细胞中铁调素(Hamp)的下调。我们假设肝损伤与 Fas 配体(在活化的 T 细胞上表达)与肝细胞上的 Fas 相互作用触发的铁稳态破坏有关。在本研究中,我们确定了 Fas 信号转导中 Flice 抑制蛋白(FLIP 长 [FLIPL])的修饰表达的影响,该蛋白干扰 Fas 信号,从而影响 Fas 引发的信号对 IL-6 和 Stat3 及其下游靶标 Hamp 的表达的影响。为了排除其他途径的可能贡献,我们使用激动性抗 Fas 抗体(而不是同种异体 T 细胞)来触发 Fas 信号。如预期的那样,通过 RNA 干扰抑制 FLIPL 不仅导致 Fas 信号应答中肝细胞凋亡增加,而且还导致 IL-6、Stat3 和 Hamp 的水平降低。相比之下,FLIPL 的过表达可保护肝细胞免受激动性抗 Fas 抗体介导的凋亡,并增加 IL-6 和 Stat3 的水平,从而以 NF-κB 依赖的方式维持 Hamp 的表达。通过液体动力学转染在体内过表达 FLIPL 同样干扰了 Fas 引发的凋亡并防止了 IL-6、Stat3 和 Hamp 的下调。这些数据表明 Fas 依赖性信号改变了铁稳态的调节,并表明 Fas 引发的信号可能导致移植前铁积累。