Method of Determining Susceptibility of a Tumor Cell to a Chemotherapeutic Agent: Novel Use of Herpes SImplex Virus

Abstract

The present invention provides a method of determining if a tumor cell is susceptible to killing by a chemotherapeutic agent, comprising: (a) providing a tumor cell; (b) infecting said tumor cell with a herpes simplex virus or a herpes simplex virus defective in an immediate early gene selected from the group consisting of ICP27, ICP4, and ICP22; and (c) determining the presence of apoptotic killing of said tumor cell, wherein the presence of apoptotic killing is indicative of susceptibility to said chemotherapeutic agent. Chemotherapeutic agent may include doxorubicin, etoposide, paclitaxel, cisplatin, or 5-fluororuacil. The present invention also provides a herpes simplex virus promoter construct having a lacZ gene to assess tumor resistance to chemotherapeutic agents.

What is claimed is: 
     
         1 . A kit for determining susceptibility of a tumor cell to apoptotic killing by a chemotherapeutic agent, said kit comprising:
 (a) a herpes simplex virus report construct, said construct comprises:
 (i) a herpes simplex virus immediate early gene promoter selected from the group consisting of ICP27, ICP4, and ICP22; and 
 (ii) a lacZ gene, 
 wherein said gene promoter is operably linked to said lacZ gene; 
   (b) β-galactosidase to detect gene expression of said lacZ gene; and   (c) an instruction, said instruction describes the use of said herpes simplex virus report construct to infect said tumor cell and staining of said β-galactosidase to detect the gene expression of said lacZ gene,
 wherein negative β-galactosidase staining reveals that said tumor cell is susceptible to chemotherapeutic agent. 
   
     
     
         2 . The kit of  claim 1 , wherein said chemotherapeutic agent is selected from doxorubicin, etoposide, paclitaxel, cisplatin, and 5-fluorouracil. 
     
     
         3 . The kit of  claim 1 , wherein said tumor cell is derived from a source selected from the group consisting of breast, brain, and cervix. 
     
     
         4 . The kit of  claim 1 , wherein said tumor cell is derived from breast. 
     
     
         5 . A kit for determining susceptibility of a tumor cell to a chemotherapeutic agent, comprising:
 (a) herpes simplex virus lacking an immediate early gene selected from the group consisting of ICP27, ICP4 and ICP22;   (b) a reagent to determine apoptosis of tumor cell; and   (c) an instruction, said instruction details the use of said herpes simplex virus in inducing apoptosis of tumor cells and measuring said apoptosis,
 wherein the presence of said apoptosis of tumor cells reveals said tumor cell is susceptible to a chemotherapeutic agent. 
   
     
     
         6 . The kit of  claim 5 , wherein said chemotherapeutic agent is selected from doxorubicin, etoposide, paclitaxel, cisplatin, and 5-fluorouracil. 
     
     
         7 . The kit of  claim 5 , wherein said tumor cell is derived from a source selected from the group consisting of breast, brain, and cervix. 
     
     
         8 . The kit of  claim 5 , wherein said tumor cell is derived from breast. 
     
     
         9 . The kit of  claim 5 , wherein said reagent determines apoptotic killing by chromatin condensation, fragmentation of nucleic, membrane blebbing or formation of apopotic bodies. 
     
     
         10 . The kit of  claim 5 , said reagent is a fluorescence probe.