SFT EXPRESSION IN A MOUSE MODEL OF HEMOCHROMATOSIS

本提案中描述的研究旨在调查遗传性血色病（HH）中观察到的异常铁积累的分子机制。 在特定目标1中，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。 由于患有HH的人类缺乏功能性HFE蛋白，因此HFE敲除小鼠是研究人类疾病的最佳可用动物模型。还将测量SFT在各种其他组织中的表达以确定SFT的表达模式。 在特定目标2中，将用SFT转染过表达HFE的HeLa细胞，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，已发现在这些细胞中铁摄取减少。 在特定目标3中，反义策略将用于阻断HeLa细胞中SFT的表达，HeLa细胞不表达可检测水平的HFE。 具体目标3将确定SFT表达的阻断是否会降低运铁蛋白介导的铁在这些细胞中的摄取。总之，这些特定的目标将有助于阐明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.