SFT EXPRESSION IN A MOUSE MODEL OF HEMOCHROMATOSIS

这项研究旨在探讨遗传性血色沉着症(HH)中铁异常积聚的分子机制。在具体目标1中，将检验HFE基因敲除小鼠肝脏和十二指肠中SFT(Stimator Of Fe Transport)基因和蛋白水平升高的假设。由于患有HH的人缺乏功能性HFE蛋白，HFE基因敲除小鼠是研究人类疾病的最佳动物模型。还将测量SFT在其他各种组织中的表达，以确定SFT的表达模式。在特定的目标2中，将SFT导入过表达HFe的HeLa细胞，并测量转铁蛋白介导的铁摄取。我们的目标是确定SFT表达的增加是否会增加转铁蛋白介导的铁摄取，而转铁蛋白介导的铁摄取在这些细胞中已被发现减少。在特定目标3中，将使用反义策略来阻断SFT在HeLa细胞中的表达，因为HeLa细胞不表达可检测到的HFE水平。特异性目标3将确定阻断SFT表达是否会减少这些细胞中转铁蛋白介导的铁摄取。综上所述，这些特定的靶点将有助于阐明HH铁代谢异常的分子机制。

The research described in this proposal aims to investigate the molecular mechanisms involved in the abnormal iron accumulation observed in the disease, hereditary hemochromatosis (HH). In Specific Aim 1, the hypothesis that levels of SFT (Stimulator of Fe Transport) mRNA and protein are increased in the liver and duodenum of HFE-knockout mice will be tested. Because humans with HH lack functional HFE protein, the HFE-knockout mouse is the best available animal model for studying the human disease. Expression of SFT in various other tissues will also be measured to determine the pattern of expression of SFT. In Specific Aim 2, HeLa cells that over-express HFE will be transfected with SFT and transferrin-mediated iron uptake will be measured. The goal is to determine if increased expression of SFT will increase transferrin-mediated iron uptake, which has been found to be decreased in these cells. In Specific Aim 3, antisense strategies will be used to block the expression of SFT in HeLa cells, which do not express detectable levels of HFE. Specific Aim 3 will determine if the blocking of SFT expression will decrease transferrin-mediated iron uptake in these cells. Together, these specific aims will be useful in elucidating the molecular mechanisms involved in the abnormal iron metabolism of HH.