Identification of genes having a role in the presentation of diabetic nephropathy

Abstract

A method for identifying a gene having a role in the presentation of diabetic nephropathy comprises culturing mesangial cells in the presence of a concentration of glucose sufficient to induce differential expression, especially up-regulation, of a gene susceptible to such differential expression and identifying the gene so induced. The cells are also optionally subjected to mechanical strain and/or TGF-β1 can be added to the culture medium. The differentially expressed genes can be identified by suppression subtractive hybridisation. The method has resulted in the identification of novel genes which play a role in the presentation of diabetic nephropathy. The genes can be used as diagnostic markers for diabetic nephropathy and as the basis of drug development programmes.

1 . A method of diagnosing and determining the progression of diabetic nephropathy which comprises, culturing mesangial cells in a medium in the presence of transforming growth factor β1 (TGF-β1) and a concentration of glucose sufficient to induce differential expression of a gene susceptible to such differential expression; 
 identifying the gene so induced by suppression subtractive hybridization; and    using said gene as a diagnostic marker for the progression and presentation of diabetic nephropathy.    
     
     
         2 . A method of identifying drugs for use in the prevention and/or therapy of diabetic nephropathy which comprises, culturing mesangial cells in a medium in the presence of transforming growth factor β1 (TGF-β1) and a concentration of glucose sufficient to induce differential expression of a gene susceptible to such differential expression; 
 identifying the gene so induced by suppression subtractive hybridization; and    screening potential drug compounds for an effect on the identified gene or on a protein encoded by said gene.    
     
     
         3 . The method according to  claim 1 , wherein the mesangial cells are cultured in the presence of a concentration of glucose sufficient to induce up-regulation of a gene susceptible to such up-regulation.  
     
     
         4 . A method according to  claim 1 , wherein the concentration of glucose is greater than 5 mM.  
     
     
         5 . A method according to  claim 1 , wherein the mesangial cells are subjected to mechanical strain.  
     
     
         6 . A method according to  claim 1 , wherein transforming growth factor β1 (TGF-β1) is added to the culture medium.  
     
     
         7 . A method according to  claim 1 , wherein the possibility of differential expression due to hyperosmolarity is excluded.  
     
     
         8 . A method according to  claim 1 , wherein the gene so differentially expressed is SEQ ID NO: 1 or SEQ ID NO:3.  
     
     
         9 . The method according to  claim 2 , wherein the mesangial cells are cultured in the presence of a concentration of glucose sufficient to induce up-regulation of a gene susceptible to such up-regulation.  
     
     
         10 . A method according to  claim 2 , wherein the concentration of glucose is greater than 5 mM.  
     
     
         11 . A method according to  claim 2 , wherein the mesangial cells are subjected to mechanical strain.  
     
     
         12 . A method according to  claim 2 , wherein transforming growth factor β1 (TGF-β1) is added to the culture medium.  
     
     
         13 . A method according to  claim 2 , wherein the possibility of differential expression due to hyperosmolarity is excluded.  
     
     
         14 . A method according to  claim 2 , wherein the gene so differentially expressed is SEQ ID NO: 1 or SEQ ID NO:3.