Tumor-targeting synthetic adenoviruses and uses thereof
Synthetic adenoviruses with liver detargeting mutations and expressing an adenovirus type 34 (Ad34) fiber protein, or a chimeric fiber protein with an Ad34 knob domain, are described. The synthetic adenoviruses traffic to sites of tumors. Use of the synthetic adenoviruses for delivering diagnostic or therapeutic transgenes to tumors are also described.
1. A method of expressing a transgene in stromal cells in a tumor, comprising infecting the stromal cells in the tumor with a synthetic adenovirus comprising:
the transgene inserted into the E1 region, wherein the transgene encodes an anti-cancer agent;
a native or modified capsid that detargets the synthetic adenovirus from the liver; and an adenovirus type 34 (Ad34) fiber protein or a chimeric fiber protein comprising an adenovirus type 5 (Ad5) shaft domain and an Ad34 knob domain,
wherein the stromal cells in the tumor express the transgene.
2. The method of claim 1 , wherein the synthetic adenovirus comprises a modified capsid that detargets the synthetic adenovirus from the liver.
3. The method of claim 1 , wherein the synthetic adenovirus further comprises one or more binding sites for a liver-specific microRNA.
4. The method of claim 3 , wherein the liver-specific microRNA is miR-122.
5. The method of claim 1 , wherein the synthetic adenovirus further comprises one or more binding sites for a spleen-specific microRNA.
6. The method of claim 5 , wherein the spleen-specific microRNA is miR142-3p.
7. The method of claim 1 , wherein expression of the transgene is regulated by a tissue-specific promoter.
8. The method of claim 1 , wherein the synthetic adenovirus comprises Ad5 capsid proteins and a chimeric fiber protein comprising an Ad5 shaft domain and an Ad34 knob domain.
9. The method of claim 1 , wherein the tumor is a pancreatic tumor or a glioblastoma.
10. The method of claim 1 , wherein the transgene encodes an anti-cancer agent selected from the group consisting of a pro-inflammatory molecule, a cytokine, an anti-angiogenic factor, an inhibitor of KRas, and an inhibitor of cytotoxic T lymphocyte-associated molecule (CTLA)-4, programmed cell death protein 1 (PD-1), carcinoembryonic antigen (CEA) and mucin 1 (MUC1).
11. The method of claim 1 , wherein the anti-cancer agent disrupts or kills the stromal cells in the tumor.
12. The method of claim 1 , wherein the transgene is operably linked to the EF1α promoter.
13. The method of claim 1 , wherein the transgene encodes a pro-inflammatory molecule or cytokine selected from the group consisting of granulocyte macrophage colony stimulating factor (GM-CSF), CD40 ligand (CD40L), Fms-related tyrosine kinase 3 (FLT3) ligand, interleukin (IL)-1b, IL-2, IL-4, IL-6, IL-12, tumor necrosis factor (TNF)-α, an interferon, a chemokine, B7-1, intercellular adhesion molecule (ICAM)-1, lymphocyte function-associated antigen (LFA)-3, transforming growth factor (TGF)-β, platelet derived growth factor (PDGF), and epidermal growth factor (EGF).
14. The method of claim 13 , wherein the transgene is operably linked to the EF1α promoter.
15. The method of claim 1 , wherein the transgene encodes an inhibitor of vascular endothelial growth factor (VEGF).
16. The method of claim 15 , wherein the transgene is operably linked to the EF1α promoter.
17. A method of reducing the size of a tumor in a subject, comprising infecting stromal cells in the tumor of the subject with a synthetic adenovirus comprising:
a therapeutic transgene inserted into the E1 region, wherein the therapeutic transgene encodes an anti-cancer agent;
a native or modified capsid that detargets the synthetic adenovirus from the liver; and an adenovirus type 34 (Ad34) fiber protein or a chimeric fiber protein comprising an adenovirus type 5 (Ad5) shaft domain and an Ad34 knob domain,
wherein the stromal cells in the tumor express the transgene, and
wherein expression of the transgene results in a reduction in the size of the tumor.
18. The method of claim 17 , wherein the therapeutic transgene encodes an anti-cancer agent selected from the group consisting of a pro-inflammatory molecule, a cytokine, an anti-angiogenic factor, an inhibitor of KRas, and an inhibitor of cytotoxic T lymphocyte-associated molecule (CTLA)-4, programmed cell death protein 1 (PD-1), carcinoembryonic antigen (CEA), and mucin 1 (MUC1).
19. The method of claim 17 , wherein the anti-cancer agent disrupts or kills the stromal cells in the tumor.
20. The method of claim 17 , wherein the transgene is operably linked to the EF1α promoter.
21. The method of claim 17 , wherein the tumor is a pancreatic tumor.
22. The method of claim 17 , wherein the tumor is a glioblastoma.
23. A synthetic adenovirus genome, comprising a nucleotide sequence at least 95% identical to SEQ ID NO: 5.
24. The synthetic adenovirus genome of claim 23 , comprising SEQ ID NO: 5.