SFT EXPRESSION IN A MOUSE MODEL OF HEMOCHROMATOSIS

本研究旨在探讨遗传性血色素沉着症（HH）中异常铁积累的分子机制。在Specific Aim 1中，我们将检验hfe敲除小鼠肝脏和十二指肠中SFT（铁转运刺激因子）mRNA和蛋白水平升高的假设。由于HH患者缺乏功能性HFE蛋白，HFE敲除小鼠是研究人类疾病的最佳动物模型。SFT在其他各种组织中的表达也将被测量，以确定SFT的表达模式。在Specific Aim 2中，过度表达HFE的HeLa细胞将被SFT转染，并测量转铁蛋白介导的铁摄取。目的是确定SFT表达的增加是否会增加转铁蛋白介导的铁摄取，在这些细胞中已发现铁摄取减少。在Specific Aim 3中，将使用反义策略来阻断HeLa细胞中SFT的表达，因为HeLa细胞不表达可检测水平的HFE。特异性Aim 3将确定阻断SFT表达是否会降低这些细胞中转铁蛋白介导的铁摄取。总之，这些特定的目的将有助于阐明HH异常铁代谢的分子机制。

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.