Microorganisms for therapy
Therapeutic methods and microorganisms therefor are provided. The microorganisms are designed to accumulate in immunoprivileged tissues and cells, such as in tumors and other proliferating tissue and in inflamed tissues, compared to other tissues, cells and organs, so that they exhibit relatively low toxicity to host organisms. The microorganisms also are designed or modified to result in leaky cell membranes of cells in which they accumulate, resulting in production of antibodies reactive against proteins and other cellular products and also permitting exploitation of proferating proliferating tissues, particularly tumors, to produce selected proteins and other products. Vaccines containing the microorganisms are provided. Combinations of the microorganisms and anti-cancer agents and uses thereof for treating cancer also are provided.
1. A method of treatment of cancer, comprising:
administering a pharmaceutical composition to a subject for treatment of cancer, wherein:
the cancer comprises a solid tumor;
treatment comprises an alleviation, reduction or amelioration of clinical symptoms or diagnostic markers associated with the tumor;
the pharmaceutical composition comprises a recombinant vaccinia virus comprising a modified thymidine kinase (TK) gene and a modified hemagglutinin (HA) gene, and an insertion, deletion or mutation in the gene or locus designated F3, wherein:
the vaccinia virus is a Lister strain;
each of the TK, HA and F3 gene or locus is inactivated; and
the F3 gene or locus occurs on the HindIII-F fragment of the vaccinia virus between open reading frames F14L and F15L.
2. The method of claim 1 , wherein the vaccinia virus is an LIVP strain.
3. The method of claim 1 , wherein the vaccinia virus has an insertion at the NotI site in the F3 locus.
4. The method of claim 1 , wherein inactivation of at least one of the genes or loci is effected by insertion of heterologous nucleic acid therein.
5. The method of claim 1 , wherein heterologous nucleic acid is inserted into the TK and HA genes to inactivate them, and heterologous nucleic acid is inserted into the F3 locus.
6. The method of claim 5 , wherein the insertion in the F3 locus is at the NotI site within the F3 gene or at a corresponding locus.
7. The method of claim 6 , wherein the insertion in the F3 locus is at position 1475 inside of the HindIII-F fragment.
8. The method of claim 5 , wherein at least one of the TK, HA gene and F3 locus comprises an insertion of heterologous nucleic acid that encodes a protein.
9. The method of claim 8 , wherein the protein is an antibody or antigen-binding fragment thereof.
10. The method of claim 8 , wherein the heterologous nucleic acid comprises a regulatory sequence operatively linked to the nucleic acid encoding the protein.
11. The method of claim 10 , wherein the regulatory sequence comprises the vaccinia virus early/late promoter p7.5.
12. The method of claim 10 , wherein the regulatory sequence comprises an early/late vaccinia pE/L promoter.
13. The method of claim 9 , wherein the virus also includes heterologous nucleic acid that encodes a detectable protein or a protein capable of inducing a detectable signal.
14. The method of claim 8 , wherein the heterologous protein is an anti-tumor agent or an anti-metastatic agent.
15. The method of claim 13 , further comprising detecting the detectable protein or detectable signal inside the subject to thereby detect tumor tissue and/or metastases and/or to monitor therapy.
16. The method of claim 8 , wherein the heterologous nucleic acid encodes a detectable protein or a protein capable of inducing a detectable signal.
17. The method of claim 16 , wherein the virus also encodes a therapeutic protein for the treatment of tumors.