Basic research in kidney-specific gene transfer using renal transplantation procedure

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The purpose of this study was to establish the conditions where we can realize the most efficient kidney-specific gene transfer using renal transplantation procedure. The approach to our goal was divided into three steps.(I) We investigated the effects of temperatures on the efficiency of gene transfer with cationic liposomes into a variety of cells derived from HeLa, LLC-PK I, normal mesangial, and normal endothelial cell lines. At a result, the following became clear : (1) The efficiency of gene transfer with cationic liposomes was almost independent of temperature ; (2) Differences of the efficiency among a variety of cells became smaller with a rise of temperature.(II) We tried to transfer reporter genes into rat kidneys which was in the convalescent stage from acute tubular necrosis (ATN) and transplanted them orthotopicafly. In advance we had treated rats with HgCl_2 to develop ATN.The kidneys were removed 10 days after transplantation and reporter gene expression was determined using RT-PCR and immunohistochemistry. At a result it was detected in all of three HgCl_2-treated rats and 5 of 6 untreated rats. In addition it was evident in glomerular and interstitial cells, occasionally tubular cells.(III) We assumed renal cystic disease as a subject of this therapy, then analyzed the molecular basis of cystogenesis and obtained new informations on it.