Vaccinia virus mutants useful for cancer immunotherapy
Disclosed herein are methods and compositions related to the treatment, prevention, and/or amelioration of cancer in a subject in need thereof. In particular aspects, the present technology relates to the use of poxviruses, including a recombinant modified vaccinia Ankara (MVA) virus or vaccinia virus with deletion of vaccinia host-range factor C7 (MVAΔC7L and VACVΔC7L, respectively), alone or in combination with immune checkpoint blocking agents, as an oncolytic and immunotherapeutic composition. In some embodiments, the technology of the present disclosure relates to a MVAΔC7L or VACVΔC7L virus further modified to express human Fms-like tyrosine kinase 3 ligand (Flt3L).
1. A method for treating a solid tumor in a subject in need thereof, the method comprising delivering to a tumor a composition comprising an effective amount of an engineered modified vaccinia Ankara (MVA) virus strain comprising a disruption of a C7L gene (MVAΔC7L), a MVAΔC7L virus genetically engineered to express hFlt3L (MVAΔC7L-hFlt3L), or combinations thereof.
2. The method of claim 1 ,
wherein the disruption comprises a deletion of the C7L gene; or
wherein the disrupted C7L gene does not encode a full-length, wild-type gene product; or
wherein the disrupted C7L gene comprises an insertion of a heterologous nucleic acid sequence into the coding sequence of the C7L gene; or
wherein the disrupted C7L gene comprises replacement of at least a portion of the gene with one or more gene cassettes; or
wherein the disrupted C7L gene comprises replacement of the entire C7L gene with one or more gene cassettes.
3. A method of stimulating an immune response against a tumor comprising:
administering to a subject an immunogenic composition comprising an engineered modified vaccinia Ankara (MVA) virus strain comprising a disruption of a C7L gene.
4. A method of stimulating an immune response against a tumor comprising:
administering to a subject an immunogenic composition comprising an engineered attenuated vaccinia virus (VACV) strain comprising a disruption of a C7L gene.
5. The method of claim 3 , wherein the tumor is melanoma, colon, breast, or prostate carcinoma.
6. The method of claim 4 , wherein the tumor is melanoma, colon, breast, or prostate carcinoma.
7. The method of claim 1 , wherein treatment comprises one or more of the following: inducing an immune response in the subject against the tumor or enhancing or promoting an ongoing immune response against the tumor in the subject, reducing the size of the tumor, eradicating the tumor, inhibiting growth of the tumor, inhibiting metastatic growth of the tumor, inducing apoptosis of tumor cells, or prolonging survival of the subject.
8. The method of claim 1 , wherein the induction, enhancement, or promotion of the immune response comprises one or more of the following:
increased levels of interferon beta (IFNB) expression in dendritic cells and THP-1 cells as compared to dendritic cells and THP-1 cells infected with the corresponding wild-type strain;
increased levels of TBK1 and IRF3 phosphorylation in dendritic cells as compared to dendritic cells infected with the corresponding wild-type strain;
increased levels of ISG expression in dendritic cells as compared to dendritic cells infected with the corresponding wild-type strain; and
increased levels of at least one of IFNB, CCL4, CCL5, and CXCL10 in tumor cells as compared to tumor cells infected with the corresponding wild-type strain.
9. The method of claim 1 , wherein the composition is administered by intratumoral or intravenous injection or a simultaneous or sequential combination of intratumoral and intravenous injection.
10. The method of claim 1 , wherein the tumor is melanoma, colon, breast, or prostate carcinoma.
11. The method of claim 1 , wherein the composition further comprises one or more immune checkpoint blocking agents.
12. The method of claim 11 , wherein the one or more immune checkpoint blocking agents is selected from the group consisting of: CTLA-4, CD80, CD86, PD-1, PDL1, PDL2, LAG3, B7-H3, B7-H4, TIM3, ICOS, II DLBCL inhibitors, BTLA, ipilimumab, nivolumab, pembrolizumab, pidilizumab, AMP-224, MPDL3280A, BMS-936559, MED14736, MSB 00107180, and any combination thereof.
13. A method for treating a solid tumor in a subject in need thereof, the method comprising delivering to a tumor a composition comprising an effective amount of an engineered attenuated vaccinia virus (VACV) strain comprising a disruption of a C7L gene (VACVΔC7L), a VACVΔC7L virus genetically engineered to express hFlt3L (VACVΔC7L-hFlt3L), or combinations thereof.
14. The method of claim 13 ,
wherein the disruption comprises a deletion of the C7L gene; or
wherein the disrupted C7L gene does not encode a full-length, wild-type gene product; or
wherein the disrupted C7L gene comprises an insertion of a heterologous nucleic acid sequence into the coding sequence of the C7L gene; or
wherein the disrupted C7L gene comprises replacement of at least a portion of the gene with one or more gene cassettes; or
wherein the disrupted C7L gene comprises replacement of the entire C7L gene with one or more gene cassettes.
15. The method of claim 13 , wherein treatment comprises one or more of the following: inducing an immune response in the subject against the tumor or enhancing or promoting an ongoing immune response against the tumor in the subject, reducing the size of the tumor, eradicating the tumor, inhibiting growth of the tumor, inhibiting metastatic growth of the tumor, inducing apoptosis of tumor cells, or prolonging survival of the subject.
16. The method of claim 13 , wherein the induction, enhancement, or promotion of the immune response comprises one or more of the following:
increased levels of interferon beta (IFNB) expression in dendritic cells and THP-1 cells as compared to dendritic cells and THP-1 cells infected with the corresponding wild-type strain;
increased levels of TBK1 and IRF3 phosphorylation in dendritic cells as compared to dendritic cells infected with the corresponding wild-type strain;
increased levels of ISG expression in dendritic cells as compared to dendritic cells infected with the corresponding wild-type strain; and
increased levels of at least one of IFNB, CCL4, CCL5, and CXCL10 in tumor cells as compared to tumor cells infected with the corresponding wild-type strain.
17. The method of claim 13 , wherein the composition is administered by intratumoral or intravenous injection or a simultaneous or sequential combination of intratumoral and intravenous injection.
18. The method of claim 13 , wherein the tumor is melanoma, colon, breast, or prostate carcinoma.
19. The method of claim 13 , wherein the composition further comprises one or more immune checkpoint blocking agents.
20. The method of claim 19 , wherein the one or more immune checkpoint blocking agents is selected from the group consisting of: CTLA-4, CD80, CD86, PD-1, PDL1, PDL2, LAG3, B7-H3, B7-H4, TIM3, ICOS, II DLBCL inhibitors, BTLA, ipilimumab, nivolumab, pembrolizumab, pidilizumab, AMP-224, MPDL3280A, BMS-936559, MED14736, MSB 00107180, and any combination thereof.