Targeting epigenetic regulators using a bacterial delivery system
One embodiment provides an attenuated Salmonella strain comprising a lysis gene or cassette operably linked to an intracellularly induced Salmonella promoter. In one embodiment, the promoter is a promoter for one of the genes in Salmonella pathogenicity island 2 type III secretion system (SPI2-T3SS) selected from the group SpiC/SsaB, SseF, SseG, SseI, SseJ, SseK1, SseK2, SifA, SifB, PipB, PipB2, SopD2, GogB, SseL, SteC, SspH1, SspH2, or SirP. In one embodiment, a Salmonella gene is under the regulation of an inducible promoter, wherein the gene is selected from ftsW, ftsA, ftsZ, murE, mukF, imp, secF, eno, hemH, tmk, dxs, uppS, cdsA, accA, pssA, msbA, tsf, trmD, cca, infB, rpoA, rpoB, rpoC, holA, dnaC, or eng. In one embodiment, the Salmonella strain further comprises a plasmid that expresses DNA, shRNA, non-coding RNA and/or a peptide.
1. An attenuated Salmonella strain comprising a lysis gene or cassette operably linked to an intracellularly induced Salmonella promoter, wherein the intracellularly induced Salmonella promoter induces lysis of the Salmonella strain when in mammalian cytosol without an extracellular inducer agent, wherein the promoter is selected from the group consisting of SseJ, SseK2, SifB, SopD2, SseL or SteC.
2. The Salmonella strain of claim 1 , further comprising a sifA deletion or wherein expression of sifA has been reduced.
3. The Salmonella strain of claim 1 , wherein the promoter is SseJ.
4. The Salmonella strain of claim 1 , wherein an essential Salmonella gene is under the regulation of an inducible promoter, wherein the gene is selected from ftsW, ftsA, ftsZ, murE, mukF, imp, secF, eno, hemH, tmk, dxs, uppS, cdsA, accA, pssA, msbA, tsf, trmD, cca, infB, rpoA, rpoB, rpoC, holA, dnaC, or eng.
5. The Salmonella strain of claim 4 , wherein the essential gene is tsf, eno, or cca.
6. The Salmonella strain of claim 1 further comprising a plasmid that expresses DNA, shRNA, non-coding RNA and/or a peptide.
7. The Salmonella strain of claim 6 , wherein the shRNA molecule is complimentary to a gene that encodes a cytoplasmic protein that promotes survival of a cancer cell.
8. The Salmonella strain of claim 6 , wherein the shRNA molecule or peptide inhibits, suppresses or blocks expression and/or activity of an epigenetic target.
9. The Salmonella strain of claim 8 , wherein the peptide comprises amino acids 143-224 of NIPP1 (SEQ ID NO:93) or 191-210 of NIPP1 (SEQ ID NO:93).
10. An attenuated Salmonella strain, wherein the Salmonella strain is VNP20009 with a sifA deletion, an intracellularly induced lysis gene or cassette, a plasmid that expresses at least one DNA, shRNA or peptide and wherein the tsf gene of the Salmonella strain is under the regulation of an inducible promoter.
11. A composition comprising the attenuated Salmonella strain claim 1 and pharmaceutically acceptable carrier.
12. A method to treat cancer comprising administering to subject in need thereof an effective amount of the attenuated Salmonella strain of claim 1 so as to treat said cancer, wherein said cancer is a solid tumor.
13. A method of inhibiting solid tumor growth/proliferation or reducing the volume/size of the tumor comprising administering to subject in need thereof an effective amount of the attenuated Salmonella strain of claim 1 so as to suppress the tumor growth or reduce the volume of the tumor.
14. A method to treat, reduce formation/number or inhibit spread of metastases from a solid tumor comprising administering to subject in need thereof an effective amount of the attenuated Salmonella strain of claim 1 so as to treat, reduce formation/number or inhibit spread of metastases from a solid tumor.
15. The method of claim 12 , wherein the cancer is selected from the group consisting of lung cancer, liver cancer, kidney cancer, breast cancer, prostate cancer, pancreatic cancer, colon cancer, head and neck cancer, ovarian cancer and gastroenterological cancer.
16. The Salmonella strain of claim 1 , wherein the lysis cassette is Lysin E from phage phiX174, the lysis cassette of phage iEPS5, or the lysis cassette from lambda phage.
17. The Salmonella strain of claim 8 , wherein the epigenetic target is at least one of EZH2, NIPP1, or PP1.
18. The Salmonella strain of claim 4 , wherein the inducible promoter is tightly regulated and induced by a small molecule that is safe to inject into humans.
19. The Salmonella strain of claim 18 , wherein the inducible promoter includes pBAD (L-arabinose), Lad (IPTG) or nahR (acetyl salicylic acid (ASA)).
20. The Salmonella strain of claim 8 , wherein the epigenetic target is at least one of EZH2, NIPP1, PP1, Suz12, EED, EZH1, RbAp48, Jarid2, YY1, CBX2, CBX4, CBX6, CBX7, PHC1, PHC2, PHC3, BMI1, PCGF2, ZNF134, RING1, RNF2, PHF1, MTF2, PHF19, SETD1A, SETD1B, CXXC1, ASH2L, DPY30, RBBP5, WDR5, KMT2A, KMT2B, KMT2C, KATE, KDM6A, NCOA6, PAGR1, PAXIP1, ASH1L, SMARCA2, SMARCA4, BPTF, or SMARCA1.