Oncolytic HSV vector
The present invention provides a recombinant herpes simplex virus (HSV), comprising (a) a mutation of the glycoprotein B (gB) at position 285 or 549, (b) a plurality of copies of one or more microRNA target sequences inserted into a locus of an HSV gene required for HSV replication, wherein said target sequence is the reverse complement of microRNA miR-124 and wherein said target sequence is present in the ICP4 gene, and (c) a transgene encoding a matrix metalloproteinase. The present invention also provides a method of killing a cancerous cell using a recombinant HSV according to the invention and a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a recombinant HSV according to the invention.
1. A recombinant herpes simplex virus (HSV), comprising (a) a mutation of the glycoprotein B (gB) at position 285 or 549, (b) a plurality of copies of one or more microRNA target sequences inserted into a locus of an HSV gene required for HSV replication, wherein said target sequence is the reverse complement of microRNA miR-124 and wherein said target sequence is present in the ICP4 gene, (c), a deletion of the internal repeat (joint) region in the HSV genome comprising one copy of the ICP0, ICP34.5, LAT, and ICP4 genes and the ICP47 promoter, and (d) a transgene that encodes a protein or polypeptide that induces patient immune response against cancer.
2. The HSV of claim 1 , wherein said microRNA target sequence(s) are inserted in the 3′ untranslated region (3′ UTR) of said HSV gene.
3. The HSV of claim 2 , wherein at least two microRNA targeting sequences are inserted.
4. The HSV of claim 1 , wherein the gB gene mutations are at both position 285 and 549.
5. The HSV of claim 4 , wherein the gB gene contains missense mutations.
6. The HSV of claim 5 , wherein the missense mutations are D285N and A549T.
7. The HSV of claim 1 , further comprising a constitutive promoter that drives the transgene.
8. The HSV of claim 7 , wherein the promoter is a cytomegalovirus (CMV) promoter.
9. A method of killing a cancerous cell, comprising exposing the cancerous cell to the HSV of claim 1 under conditions sufficient for said HSV to infect said cancerous cell, whereby replication of the HSV within the cancerous cell results in cell death.
10. The method of claim 9 , wherein the cell is in vivo.
11. The method of claim 9 , wherein the cell is within a tumor.
12. The method of claim 11 , wherein the tumor is glioblastoma multiforme.
13. The method of claim 9 , wherein the cell is human.
14. The method of claim 11 , wherein the tumor is within the brain of an animal.
15. The method of claim 14 , wherein the HSV is exposed to the cell by intracranially injecting the HSV into the animal.
16. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and the HSV of claim 1 .
17. The HSV of claim 1 , which comprises a transgene encoding a matrix metalloproteinase.
18. The HSV of claim 17 , wherein the matrix metalloproteinase is matrix metalloproteinase 9.