Oncolytic viral vectors and uses thereof
Oncolytic viral vectors that incorporate one or more of the following features: viral replication restriction by insertion of 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. Such viral vectors can be used for the treatment and prevention of cancer.
1. A recombinant oncolytic herpes simplex virus (HSV) comprising:
(i) a first microRNA (miRNA) target sequence cassette (miR-TS cassette) inserted into a first viral gene and comprising at least 2 target sequences for each of miR-124, miR-1, and miR-143;
(ii) a second miR-TS cassette inserted into a second viral gene and comprising at least 2 target sequences for each of miR-128, miR-219a, and miR-122; and
(iii) a third miR-TS cassette inserted into a third viral gene and comprising at least 2 target sequences for each of miR-219a, miR-204, and miR-128;
and wherein:
(a) the first miR-TS cassette comprises a nucleic acid sequence that is at least 95% identical to SEQ ID NO: 852;
(b) the second miR-TS cassette comprises a nucleic acid sequence that is at least 95% identical to SEQ ID NO: 853; or
(c) the third miR-TS cassette comprises a nucleic acid sequence that is at least 95% identical to SEQ ID NO: 854.
2. The recombinant HSV of claim 1 , further comprising a fourth miR-TS cassette inserted into a fourth viral gene and comprising at least 2 target sequences for each of miR-137, miR-217, and miR-126, and wherein the fourth miR-TS cassette comprises a nucleic acid sequence that is at least 95% identical to SEQ ID NO: 855.
3. The recombinant HSV of claim 1 , wherein each of the miR-TS cassettes comprises 4 target sequences for each of the respectively miRNAs.
4. The recombinant HSV of claim 1 , wherein the first viral gene comprises at least one ICP4.
5. The recombinant HSV of claim 1 , wherein the second viral gene is ICP27.
6. The recombinant HSV of claim 1 , wherein the third viral gene comprises at least one ICP34.5.
7. The recombinant HSV of claim 2 , wherein the fourth viral gene is UL8.
8. The recombinant HSV of claim 2 , wherein the first miR-TS cassette comprises a nucleic acid sequence that is at least 95% identical to SEQ ID NO: 852.
9. The recombinant HSV of claim 8 , wherein the second miR-TS cassette comprises a nucleic acid sequence that is at least 95% identical to SEQ ID NO: 853.
10. The recombinant HSV of claim 9 , wherein the third miR-TS cassette comprises a nucleic acid sequence that is at least 95% identical to SEQ ID NO: 854.
11. The recombinant HSV of claim 1 , further comprising a heterologous polynucleotide sequence encoding one or more payload molecules.
12. The recombinant HSV of claim 11 , wherein the heterologous polynucleotide sequence encodes a payload selected from the group consisting of IL-12, CCL4, and CXCL10.
13. The recombinant HSV of claim 11 , wherein the heterologous polynucleotide sequence encodes two or more payloads selected from the group consisting of IL-12, CCL4, and CXCL10.
14. A viral stock comprising the oncolytic HSV of claim 1 .
15. A composition comprising the oncolytic HSV of claim 1 and a pharmaceutically-acceptable carrier.
16. A method of killing a cancerous cell, comprising exposing the cancerous cell to the oncolytic HSV of claim 1 under conditions sufficient for the oncolytic HSV to infect and replicate within said cancerous cell, and wherein replication of the oncolytic HSV within the cancerous cell results in cell death.
17. The method of claim 16 , wherein the cancerous cell has a reduced expression of a miR capable of binding to the one or more miR-target sequences compared to the expression of the miR in a non-cancerous cell.
18. The method of claim 16 , wherein replication of the oncolytic virus is increased or maintained in cancerous cells with a reduced expression of the miR capable of binding to the one or more miR-target sequences.
19. The method of claim 16 , wherein the cell is in vivo.
20. The method of claim 19 , wherein the cell is within a tumor.
21. A method of treating cancer in a subject in need thereof, comprising administering the oncolytic virus of claim 1 to a subject in need thereof.
22. The method of claim 21 , wherein the subject is a mouse, a rat, a rabbit, a cat, a dog, a horse, a non-human primate, or a human.
23. The method of claim 21 , wherein the oncolytic virus or compositions thereof are administered intravenously, subcutaneously, intratumorally, intramuscularly, or intranasally.
24. The method of claim 21 , wherein the cancer is selected from lung cancer, breast cancer, ovarian cancer, cervical cancer, prostate cancer, testicular cancer, colorectal cancer, colon cancer, pancreatic cancer, liver cancer, gastric cancer, head and neck cancer, thyroid cancer, malignant glioma, glioblastoma, melanoma, B-cell chronic lymphocytic leukemia, diffuse large B-cell lymphoma (DLBCL), and marginal zone lymphoma (MZL).
25. The method of claim 24 , wherein the lung cancer is small cell lung cancer or non-small cell lung cancer.
26. The method of claim 24 , wherein the liver cancer is hepatocellular carcinoma (HCC).