Identification of mutations in herpes simplex virus envelope glycoproteins that enable or enhance vector retargeting to novel non-hsv receptors

Abstract

The present invention provides an HSV vector comprising an envelope comprising one or more mutant gB and/or gH envelope glycoproteins, whereby the HSV vector exhibits at least 25 % increased rate-of-entry after 20 minutes when assayed at 30 ° C. or 37 ° C. in Vero cells after first incubating at 4 ° C. relative to a control HSV comprising wild-type gB and gH glycoproteins. The invention also provides a viral stock comprising the inventive HSV vector.

1 . An HSV vector comprising an envelope comprising one or more mutant gB and/or gH envelope glycoproteins, whereby the HSV vector exhibits at least 25% increased rate-of-entry after 20 minutes when assayed at 30° C. in Vero cells after first incubating at 4° C. relative to a control HSV comprising wild-type gB and gH glycoproteins. 
     
     
         2 . The HSV vector of  claim 1 , further comprising a non-native ligand capable of specifically binding a surface component of a predetermined cell type. 
     
     
         3 . The HSV vector of  claim 2 , wherein the predetermined cell type is a cancer cell. 
     
     
         4 . The HSV vector of  claim 2 , wherein the ligand is incorporated into a viral envelope glycoprotein of the HSV vector. 
     
     
         5 . The HSV vector of  claim 1 , wherein at least one additional viral envelope glycoprotein of the HSV vector is impaired for binding to its natural receptor or is deleted. 
     
     
         6 . The HSV vector of  claim 1 , comprising a genome comprising an exogenous expression cassette. 
     
     
         7 . The HSV of  claim 6 , wherein the expression cassette encodes an agent that enhances tumor killing activity. 
     
     
         8 . The HSV vector of  claim 1 , comprising a genome comprising a target sequence for a cellular microRNA. 
     
     
         9 . The HSV vector of  claim 1 , which is a human HSV-1 vector. 
     
     
         10 . A viral stock comprising the HSV vector of  claim 1 . 
     
     
         11 . An HSV vector comprising an envelope comprising one or more mutant gB and/or gH envelope proteins, whereby the HSV vector exhibits at least 25% increased rate-of-entry after 20 minutes when assayed at 37° C. in Vero cells after first incubating at 4° C. relative to a control HSV comprising wild-type gB and gH glycoproteins. 
     
     
         12 . The HSV vector of  claim 11 , further comprising a non-native ligand capable of specifically binding a surface component of a predetermined cell type. 
     
     
         13 . The HSV vector of  claim 12 , wherein the predetermined cell type is a cancer cell. 
     
     
         14 . The HSV vector of  claim 12 , wherein the ligand is incorporated into a viral envelope glycoprotein of the HSV vector. 
     
     
         15 . The HSV vector of  claim 11 , wherein at least one additional viral envelope glycoprotein of the HSV vector is impaired for binding to its natural receptor or is deleted. 
     
     
         16 . The HSV vector of  claim 11 , comprising a genome comprising an exogenous expression cassette. 
     
     
         17 . The HSV of  claim 16 , wherein the expression cassette encodes an agent that enhances tumor killing activity. 
     
     
         18 . The HSV vector of  claim 11 , comprising a genome comprising a target sequence for a cellular microRNA. 
     
     
         19 . The HSV vector of  claim 11 , which is a human HSV-1 vector. 
     
     
         20 . A viral stock comprising the HSV vector of  claim 11 .