Oncolytic herpes simplex virus and therapeutic uses thereof

Abstract

The present invention relates to variants of herpes simplex virus (HSV) that selectively infect and replicate in cancer cells, including HSV strains that selectively infect and replicate in bladder cancer cells. Preferred HSV of the invention have intact endogenous Us11 and Us12 genes and have genes encoding ICP34.5 replaced with a gene encoding Us11 fused to an HSV immediate early (IE) promoter. The variant HSV of the invention also comprise one or more additional heterologous genes encoding immunomodulatory polypeptides. Methods and compositions using these variant HSV, for example, for treating cancer in a subject, are also provided.

What is claimed is: 
     
         1 . A variant herpes simplex virus (HSV) having an intact endogenous Us12 encoding gene and an intact endogenous Us11 encoding gene, lacking functional ICP34.5 encoding genes,
 wherein each ICP34.5 encoding gene is replaced by a polynucleotide cassette comprising:   (a) a Us11 encoding gene operably associated with an immediate early (IE) promoter; and   (b) at least one heterologous gene encoding a polypeptide capable of enhancing an anti-tumor response.   
     
     
         2 . The variant HSV according to  claim 1 , in which said at least one heterologous gene encodes an immunomodulatory polypeptide. 
     
     
         3 . The variant HSV according to  claim 2 , in which the immunomodulatory polypeptide is selected from the group consisting of a TAP 1/2 (“TAP”) inhibitor, granulocyte macrophage colony stimulating factor (GM-CSF), tumor necrosis factor (TNF)-alpha and CD40 ligand (CD40L). 
     
     
         4 . The variant HSV according to  claim 3 , in which said at least one immunomodulatory polypeptide is GM-CSF. 
     
     
         5 . The variant HSV according to  claim 1 , in which said at least one heterologous gene encodes a prodrug converting enzyme. 
     
     
         6 . The variant HSV according to  claim 1 , comprising at least two (2) heterologous genes encoding a polypeptide capable of enhancing an anti-tumor response. 
     
     
         7 . The variant HSV according to  claim 6 , wherein said one of said two heterologous genes encodes a TAP inhibitor and the other encodes a mammalian GM-CSF. 
     
     
         8 . The variant HSV according to  claim 7 , in which the TAP inhibitor inhibits a non-human TAP. 
     
     
         9 . The variant HSV according to  claim 8 , in which the TAP inhibitor inhibits a murine TAP. 
     
     
         10 . The variant HSV according to  claim 9 , in which the TAP inhibitor is the UL49.5 polypeptide from bovine herpesvirus. 
     
     
         11 . The variant HSV according to  claim 6 , wherein one of said two heterologous genes encodes a TAP inhibitor and the other encodes a prodrug converting enzyme. 
     
     
         12 . A variant herpes simplex virus (HSV) having an intact endogenous Us12 encoding gene and an intact endogenous Us11 encoding gene, lacking functional ICP34.5 encoding genes,
 wherein each ICP34.5 encoding gene is replaced by a polynucleotide cassette comprising:   (a) a Us11 encoding gene operably associated with an immediate early (IE) promoter; and   (b) a gene encoding an inhibitor of antigen presentation on class I major histocompatibility complex (MHC) molecules, wherein said inhibitor is capable of inhibiting antigen presentation on the surface of virally infected tumor cells.   
     
     
         13 . The variant HSV according to  claim 12 , wherein said inhibitor of antigen presentation is a TAP inhibitor. 
     
     
         14 . The variant HSV according to  claim 12 , in which the polynucleotide cassette additionally comprises a heterologous gene encoding a polypeptide capable of enhancing an anti-tumor response. 
     
     
         15 . The variant HSV according to  claim 14 , in which the heterologous gene encodes GM-CSF. 
     
     
         16 . The variant HSV according to  claim 1 , in which the IE promoter is an α27 IE promoter. 
     
     
         17 . The variant HSV according to  claim 1 , in which the heterologous gene is operably associated with a promoter selected from the group consisting of a CMV promoter and an EF1α promoter. 
     
     
         18 . The variant HSV according to  claim 13 , wherein said TAP inhibitor is a bovine herpesvirus (BHV) UL49.5 polypeptide. 
     
     
         19 . A variant herpes simplex virus (HSV) having an intact endogenous Us12 encoding gene and an intact endogenous Us11 encoding gene, lacking functional ICP34.5 encoding genes, wherein each ICP34.5 encoding gene is replaced with a polynucleotide cassette comprising a nucleic acid sequence set forth in one of SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24 and SEQ ID NO: 25. 
     
     
         20 . A variant herpes simplex virus (HSV) having a genome sequence set forth in SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, or SEQ ID NO: 30. 
     
     
         21 . A pharmaceutical formulation comprising: the variant HSV according to  claim 1 , and a pharmaceutically acceptable carrier for administration to tumor cells. 
     
     
         22 . The pharmaceutical formulation according to  claim 21 , wherein said administration to tumor cells is in situ. 
     
     
         23 . The pharmaceutical formulation according to  claim 21 , wherein said virus selectively infects human bladder cancer cells, human melanoma cells, human ovarian cancer cells, or human glioblastoma cells. 
     
     
         24 . A method for killing tumor cells in a subject comprising: administering to a subject in need thereof the pharmaceutical formulation according to  claim 21  under conditions effective to kill tumor cells in the subject. 
     
     
         25 . The method according to  claim 24 , wherein the tumor cells are selected from the group consisting of astrocytoma, oligodendroglioma, meningioma, neurofibroma, glioblastoma, ependymoma, Schwannoma, neurofibrosarcoma, medulloblastoma, melanoma cells, pancreatic cancer cells, prostate carcinoma cells, breast cancer cells, lung cancer cells, colon cancer cells, hepatoma cells, mesothelioma, bladder cancer cells, and epidermoid carcinoma cells. 
     
     
         26 . The method according to  claim 24 , wherein said virus selectively replicates in human bladder cancer cells, melanoma cells, ovarian cancer cells, or glioblastoma cells. 
     
     
         27 . The method according to  claim 24 , wherein the subject is a mammal. 
     
     
         28 . The method according to  claim 24 , wherein the mammal is a human. 
     
     
         29 . The method according to  claim 24 , wherein the administration is carried out by injection, infusion, instillation or inhalation. 
     
     
         30 . A method for treating cancer, wherein said method comprises administering to an individual in need of treatment, a therapeutically effective amount of the pharmaceutical formulation according to  claim 24 . 
     
     
         31 . The method according to  claim 30 , wherein the individual is a mammal. 
     
     
         32 . The method according to  claim 31 , wherein the mammal is a human. 
     
     
         33 . The method according to  claim 30 , wherein the cancer is selected from the group consisting of bladder cancer, melanoma, ovarian cancer and glioblastoma.