Oncolytic viral vectors and uses thereof

Abstract

The present disclosure relates to recombinant viral vectors for the treatment and prevention of cancer. Oncolytic viral vectors incorporate one or more of the following features: viral replication restriction by insertion of tumor-suppressive microRNA (miRNA) target sequences into the viral genome; disruption of oncogenic miRNA function; cancer microenvironment remodeling; and cancer cell targeting by incorporation of protease-activated antibodies into the viral particle.

1 .- 97 . (canceled) 
     
     
         98 . A recombinant herpes simplex virus (HSV) comprising a target sequence for a first microRNA (miR) and a target sequence for a second miR, wherein an ICP4 locus of the recombinant HSV genome comprises the target sequence for the first miR, and wherein an ICP27 locus of the recombinant HSV genome comprises the target sequence for the second miR. 
     
     
         99 . The recombinant HSV of  claim 98 , wherein the first miR is miR-124. 
     
     
         100 . The recombinant HSV of  claim 98 , wherein the second miR is miR-122. 
     
     
         101 . The recombinant HSV of  claim 98 , wherein the target sequence for the first miR is incorporated into the 5′ untranslated region (UTR) or 3′ UTR of the ICP4 locus, and wherein the target sequence for the second miR is incorporated into the 5′ UTR or 3′ UTR of the ICP27 locus. 
     
     
         102 . The recombinant HSV of  claim 98 , wherein two or more copies of the target sequences for the first miR are inserted into the ICP4 locus. 
     
     
         103 . The recombinant HSV of  claim 98 , wherein two or more copies of the target sequences for the second miR are inserted into the ICP27 locus. 
     
     
         104 . The recombinant HSV of  claim 98 , wherein the ICP4 locus comprises target sequences for one or more miRs other than the first and the second miRs. 
     
     
         105 . The recombinant HSV of  claim 98 , wherein the ICP27 locus comprises target sequences for one or more miRs other than the first and the second miRs. 
     
     
         106 . The recombinant HSV of  claim 98 , wherein replication of the recombinant HSV is reduced or attenuated in a normal cell compared to replication of the recombinant HSV in a cancerous cell. 
     
     
         107 . The recombinant HSV of  claim 106 , wherein replication of the recombinant HSV in the normal cell is reduced by at least 50% compared to replication of the recombinant HSV in the cancerous cell. 
     
     
         108 . The recombinant HSV of  claim 107 , wherein the cancer of the cancerous cell is selected from the group consisting of colorectal cancer, pancreatic cancer, breast cancer, liver cancer, oral cancer, ovarian cancer, prostate cancer, brain cancer, cervical cancer, and lung cancer. 
     
     
         109 . The recombinant HSV of  claim 98 , comprising a heterologous polynucleotide sequence encoding a protein selected from a cytokine and an anti-immunosuppressive protein. 
     
     
         110 . The recombinant HSV of  claim 109 , wherein the protein comprises IL-12. 
     
     
         111 . The recombinant HSV of  claim 109 , wherein the protein comprises an anti-PD1 protein and/or an anti-CTLA4 protein. 
     
     
         112 . The recombinant HSV of  claim 109 , wherein the heterologous polynucleotide sequence is inserted into the recombinant HSV genome between the UL3 and UL4 open reading frames. 
     
     
         113 . The recombinant HSV of  claim 98 , comprising a deletion of one copy of each of the ICP0, ICP34.5, LAT, and ICP4 genes in the internal joint region. 
     
     
         114 . A nucleic acid molecule encoding the recombinant HSV of  claim 98 . 
     
     
         115 . A viral stock comprising the recombinant HSV of  claim 98 . 
     
     
         116 . A composition comprising the recombinant HSV of  claim 98  and a pharmaceutically acceptable carrier. 
     
     
         117 . A method of killing a cancerous cell, comprising exposing a cancerous cell to the recombinant HSV of  claim 98  under conditions sufficient for the recombinant HSV to infect and replicate within said cancerous cell, and wherein replication of the recombinant HSV within the cancerous cell results in cell death. 
     
     
         118 . A method of treating a cancer in a subject in need thereof, comprising administering the recombinant HSV of  claim 98  to the subject. 
     
     
         119 . The method of  claim 118 , wherein the subject is a mouse or a human. 
     
     
         120 . The method of  claim 118 , wherein the recombinant HSV is administered intravenously, subcutaneously, intratumorally, or intramuscularly. 
     
     
         121 . The method of  claim 118 , wherein the cancer is colorectal cancer, pancreatic cancer, breast cancer, liver cancer, oral cancer, ovarian cancer, prostate cancer, brain cancer, cervical cancer, or lung cancer.