Genetically-engineered newcastle disease virus as an oncolytic agent, and methods of using same

Abstract

Recombinant strains of avian paramyxovirus (APMV), such as Newcastle disease virus (NDV), are provided. Also provided are compositions comprising them, and methods of using them to lyse tumor cells and to treat cancer. In certain aspects, genetically-engineered viral strains that incorporate therapeutic transgenes are also provided. The recombinant viruses may be used in accordance with methods of providing enhanced oncolytic efficacy and delivering an oncolytic virus to tumors present in a patient. Also provided are methods for identifying a recombinant virus as an oncolytically-effective agent.

1 . A recombinant Newcastle disease virus (rNDV) strain, wherein said rNDV has been genetically-modified to include one or more transgenes that induce apoptosis in one or more tumor cell lines, selected from the group consisting of transgenes that induce production of pro-apoptotic proteins, transgenes that activate tumor suppressor genes; and transgenes that activate pro-apoptotic proteins. 
     
     
         2 . The rNDV of  claim 1 , wherein said rNDV is selected from the group consisting of recombinant Beaudette C and recombinant LaSota. 
     
     
         3 . The rNDV of  claim 1 , wherein said one or more rNDV strains comprises cloned cDNA from a naturally-occurring NDV. 
     
     
         4 . The rNDV of  claim 1 , wherein said rNDV is genetically-modified to exhibit reduced pathogenicity in a natural host as compared to a non-genetically-modified rNDV. 
     
     
         5 . The rNDV of  claim 1 , wherein said transgenes that induce production of pro-apoptotic proteins in tumor cells are selected from the group consisting of cytokines, fas ligands, FADD, caspases-8, -9, -3, Smac/DIABLO, cytochrome c, IFN-beta, RANTES, IP-10, TNF-alpha, CD95 ligands, and TRAIL. 
     
     
         6 . The rNDV of  claim 1 , wherein said transgenes that activate tumor suppressing genes in tumor cells encode p53. 
     
     
         7 . The rNDV of  claim 1 , wherein said transgenes that activate tumor suppressing proteins in tumor cells activate proteins selected from the group consisting of caspase-8, caspase-9, caspase-3, Smac/DIABLO, and cytochrome c. 
     
     
         8 . The rNDV of  claim 1 , wherein said rNDV comprises one or more reporter genes selected from the group consisting of green fluorescent protein (GFR) and luciferase. 
     
     
         9 . The rNDV of  claim 1 , wherein said rNDV comprises one or more genetic modifications to influence the ability of an rNDV-infected tumor cell line to replicate in vitro. 
     
     
         10 . The rNDV of  claim 1 , wherein said one or more tumor cell lines are selected from the group consisting of tumor cells of neuroectodermal, mesenchymal, and epithelial origin. 
     
     
         11 . The rNDV of  claim 1 , wherein said rNDV activates a mitochondrial cell death pathway in said tumor cell line. 
     
     
         12 . The rNDV of  claim 1 , wherein said rNDV activates a death receptor-dependent apoptosis pathway in said tumor cell line. 
     
     
         13 . A composition comprising one or more rNDV strains of  claim 1 , and one or more pharmaceutically-acceptable excipients. 
     
     
         14 . The composition of  claim 13 , further comprising a chemotherapeutic agent. 
     
     
         15 . The composition of  claim 13 , further comprising an immunomodulatory agent. 
     
     
         16 . A method for lysing tumor cells, comprising providing an oncolytically-effective amount of one or more of the rNDV strains of  claim 1  directly to said tumor cells. 
     
     
         17 . The method of  claim 16 , wherein said tumor cells are lysed in vitro. 
     
     
         18 . The method of  claim 16 , wherein the oncolytically-effective amount of one or more rNDV strains is administered directly into a tumor found in a patient. 
     
     
         19 . A method for treating cancer in a patient suffering therefrom, comprising the step of administering directly into a tumor present in said patient a therapeutically-effective amount of a composition comprising one or more of the rNDV strains of  claim 1 . 
     
     
         20 . A method for identifying an rNDV strain that is oncolytically-effective against a tumor cell line, comprising the steps of:
 isolating a tumor cell line from a tumor;   infecting said tumor cell line with an rNDV strain; and   detecting that said rNDV has induced apoptosis in said tumor cell line by conducing an assay for one or more metabolic indicators of cell death.