Recombinant cardiomyocytes and cardiomyocyte cell lines expressing herg

Abstract

The present disclosure relates generally to recombinant cardiomyocytes and cardiomyocyte cell lines overexpressing hERG and uses thereof.

1 . A recombinant cell line comprising recombinant cardiomyocytes stably expressing hERG. 
     
     
         2 . The recombinant cell line of  claim 1 , wherein the recombinant cardiomyocytes comprise a transduced nucleic acid sequence encoding hERG. 
     
     
         3 . The recombinant cell line of  claim 1 , wherein hERG comprises an amino acid sequence as set forth in amino acids 1-1159 of SEQ ID NO: 1. 
     
     
         4 . The recombinant cell line of  claim 1 , wherein the cell line is designated hMYO-hERG (ATCC Designation No. PTA-123324). 
     
     
         5 . The recombinant cell line of  claim 1 , wherein the recombinant cardiomyocytes are progeny, descendants or derivatives of hMYO-hERG (ATCC Designation No. PTA-123324). 
     
     
         6 . A stable cardiomyocyte cell line overexpressing hERG comprising recombinant cardiomyocytes that are progeny, descendants or derivatives of hMYO-hERG (ATCC Designation No. PTA-123324). 
     
     
         7 . A method of preparing the cell line according to  claim 1 , comprising:
 transfecting or transducing human cardiomyocytes with a nucleic acid sequence encoding hERG; and   selecting the cardiomyocytes stably expressing hERG.   
     
     
         8 . The method of  claim 7 , wherein the transfecting or transducing is with a vector comprising the nucleic acid sequence. 
     
     
         9 . The method of  claim 8 , wherein the vector is a retroviral vector. 
     
     
         10 . The method of  claim 9 , wherein the vector is a lentiviral vector. 
     
     
         11 . The method of  claim 10 , further comprising generating pseudo-lentiviral particles. 
     
     
         12 . A method for determining cardiotoxicity of a compound using the cell line of  claim 1 . 
     
     
         13 . A method of screening compounds for hERG inhibitory activity comprising using the cell line of  claim 1 . 
     
     
         14 . A method for determining the activity of a compound to inhibit hERG comprising using the cell line of  claim 1 . 
     
     
         15 . A method for determining the activity of a compound to inhibit hERG comprising: a) providing recombinant cardiomyocytes overexpressing hERG; b) contacting the cardiomyocytes with the compound; c) measuring a test current; and d) determining if the test current is reduced in the presence of the compound, wherein a reduced test current is indicative of hERG inhibitory activity. 
     
     
         16 . The method of  claim 15 , wherein the test current is measured with electrophysiology techniques. 
     
     
         17 . The method of  claim 15 , wherein the test current is measured with a patch clamp apparatus. 
     
     
         18 . The method of  claim 15 , wherein the test current is compared before and after contacting the recombinant cardiomyocytes with the compound. 
     
     
         19 . The method of  claim 15  wherein the recombinant cardiomyocytes are derived from the cell line designated hMYO-hERG (ATCC Designation No. PTA-123324). 
     
     
         20 . The method of  claim 15 , wherein the recombinant cardiomyocytes are progeny, descendants or derivatives of hMYO-hERG (ATCC Designation No. PTA-123324). 
     
     
         21 . A method for determining the ability of a compound to reduce cell viability comprising using the cell line of  claim 1 . 
     
     
         22 . A method for determining the activity of a compound to reduce cell viability comprising: a) providing recombinant cardiomyocytes overexpressing hERG; b) contacting the cardiomyocytes with the compound in the presence of a viability indicator compound; c) measuring a signal of the indicator compound; and d) determining if the signal is reduced or increased in the presence of the compound, wherein a reduced or increased signal is indicative of reduced cell viability. 
     
     
         23 . The method of  claim 22 , wherein the indicator compound is an indicator dye. 
     
     
         24 . The method of  claim 22 , wherein the signal of the indicator compound is an absorbance, luminescence or fluorescence signal. 
     
     
         25 . The method of  claim 22 , wherein the signal is compared before and after contacting the cardiomyocytes with the compound. 
     
     
         26 . The method of  claim 22  wherein the recombinant cardiomyocytes are from the cell line designated hMYO-hERG (ATCC Designation No. PTA-123324). 
     
     
         27 . The method of  claim 22 , wherein the recombinant cardiomyocytes are progeny, descendants or derivatives of hMYO-hERG (ATCC Designation No. PTA-123324). 
     
     
         28 . A kit comprising the recombinant cardiomyocytes from the cell line of  claim 1 .