RNA bioassay

Abstract

The present invention relates to methods for evaluating the cell damaging potential of an agent by determining the ability of the agent to increase messenger RNA release in cells.

1 . A method for evaluating whether an agent causes cell damage comprising: treating one or more cells with an agent; and measuring the effect of said agent on the amount of RNA released by said cells.  
     
     
         2 . A method of  claim 1  wherein said cells comprise one or more cells of mammalian origin.  
     
     
         3 . A method of  claim 2  wherein said cells comprise one or more cells of human origin.  
     
     
         4 . A method of  claim 1  wherein said measurement comprises measuring the effect of said agent on the quantity of one or more RNA markers released by said cells.  
     
     
         5 . A method of  claim 4  wherein said cells are of mammalian liver origin and said one or more RNA markers comprise albumin.  
     
     
         6 . A method of  claim 4  wherein said cells are of mammalian heart origin and said one or more RNA markers comprise one or more of troponin T and cardiac actin.  
     
     
         7 . A method of  claim 4  wherein said cells are of mammalian smooth muscle origin and said one or more RNA markers comprise one or more of SM22 and smooth muscle actin.  
     
     
         8 . A method of  claim 4  wherein said cells are of mammalian kidney origin and said one or more RNA markers comprise one or more of Tamm-Horsfall protein and Aquaporin-2.  
     
     
         9 . A method of  claim 1  further comprising characterizing said agent as an agent that is likely to cause cell injury, wherein said injury is characterized by increased RNA release, provided the amount of RNA released by said cells has increased by at least two fold.  
     
     
         10 . A method of any one of  claim 1  to  9  wherein said measurement comprises measuring RNA levels by RT-PCR.  
     
     
         11 . A method of evaluating the cell-protective characteristics of an agent comprising: treating one or more cells with a first agent that causes cell injury, wherein said injury is characterized by increased RNA release from the injured cells; treating said cell with a second agent; and measuring the effect of said second agent in reducing the amount of RNA release by said cells that is caused by said first agent.  
     
     
         12 . A method of evaluating the effect of an agent on a subject comprising: treating a mammalian subject with an agent; and measuring the effect of said treatment in increasing release into the blood of one or more RNA markers.  
     
     
         13 . A method of  claim 12  wherein said subject is a human.  
     
     
         14 . A method of  claim 13  wherein said gene markers are selected from one or more of: albumin, cardiac actin, troponin T, SM22, smooth muscle actin, kidney androgen specific protein, Tamm-Horsfall protein and aquaporin-2.  
     
     
         15 . A method of  claim 14  wherein said gene markers are selected from one or more of: albumin, cardiac actin, troponin T, SM22, smooth muscle actin, kidney androgen specific protein, Tamm-Horsfall protein and aquaporin-2.  
     
     
         16 . A method of any one of  claims 12  to  15  wherein the measurement of the quantity of RNA comprises drawing blood from said subject and measuring RNA levels in said blood using RT-PCR.  
     
     
         17 . A method for diagnosing a pathological state of the liver comprising, obtaining a blood sample from a mammalian subject and measuring one or more liver RNA markers.  
     
     
         18 . A method of  claim 17  wherein said liver RNA markers comprise albumin.  
     
     
         19 . A method of  claim 18  wherein said mammalian subject is a human.  
     
     
         20 . A method of characterizing the pathological state of the liver of a subject comprising, obtaining a blood sample from a mammalian subject, measuring a liver RNA marker in said blood sample, and characterizing said subject as having a pathological state of the liver, provided said blood sample contains at least a two fold higher amount of said liver RNA marker than that which is measured in a control subject having no pathological state of the liver.