ENHANCED CANCER TREATMENT AND MONITORING USING RECOMBINANT VECTORS
This disclosure relates to replication competent viral vectors for treating cell proliferative disorders. The disclosure further relates to the use of such replication competent viral vectors for delivery and expression of a heterologous nucleic acid in normal and diseased tissues and methods and compositions that facilitate such delivery and expression to tissues in vivo and in vitro. The disclosure further relates to replication competent retroviral vectors for these uses and in conjunction with methods and compositions that facilitate in vivo therapeutics.
1 - 18 . (canceled)
19 . A method of treating a subject with a cancer, comprising:
administering to the subject a replicative viral vector comprising a polynucleotide encoding a polypeptide having cytosine deaminase activity;
administering 5-fluorocytosine (5-FC) to the subject, wherein the polypeptide converts 5-FC to 5-fluorouracil (5-FU); and
subjecting the subject to radiation therapy so as to cause death of cancer cells wherein the cancer cells are more sensitive to radiation therapy by having local concentrations of 5-FU.
20 . The method of claim 19 , wherein an inhibitor of pattern recognition receptor (PRR) or interferon (IFN) signaling pathways is administered to the subject at the same time as the replicative viral vector is administered to the subject.
21 . The method of claim 19 , wherein an inhibitor of PRR or IFN signaling pathways is administered to the subject within a period of one day before to 2 days after administration of the replicative viral vector to the subject.
22 . The method of claim 21 , wherein the inhibitor of IFN signaling is an anti-IFN antibody or anti-IFN receptor antibody.
23 . The method of claim 19 , wherein the replicative viral vector is a recombinant replicative retrovirus or a recombinant replicative retroviral vector.
24 . The method of claim 23 , wherein one or more doses of the recombinant replicative retrovirus is administered locally at the site of the cancer.
25 . The method of claim 23 , wherein one or more doses of the recombinant replicative retrovirus is administered intravenously.
26 . The method of claim 25 , wherein each dose of the one or more doses of the recombinant replicative retrovirus comprises from 10 5 to 10 12 transducing units (TU) of the recombinant replicative retrovirus.
27 . The method of claim 23 , wherein the replication competent retrovirus comprises:
a retroviral GAG protein;
a retroviral POL protein;
a retroviral envelope;
a retroviral polynucleotide comprising Long-Terminal Repeat (LTR) sequences at the 3′ end of the retroviral polynucleotide sequence, a promoter sequence at the 5′ end of the retroviral polynucleotide, said promoter being suitable for expression in a mammalian cell, a gag nucleic acid domain, a pol nucleic acid domain and an env nucleic acid domain;
a cassette comprising an internal ribosome entry site (IRES) operably linked to a heterologous polynucleotide that encodes a polypeptide having cytosine deaminase activity, wherein the cassette is positioned 5′ to the 3′ LTR and 3′ to the env nucleic acid domain encoding the retroviral envelope; and
cis-acting sequences necessary for reverse transcription, packaging and integration into a neoplastic cell.
28 . The method of claim 27 , wherein the retroviral polynucleotide sequence is derived from a murine leukemia virus (MLV), a Moloney murine leukemia virus (MoMLV), a Feline leukemia virus (FeLV), or a Gibbon ape leukemia virus (GALV).
29 . The method of claim 27 , wherein the retroviral envelope is an amphotropic MLV envelope.
30 . The method of claim 19 , wherein the polynucleotide comprises a cytosine deaminase sequence that has been human codon optimized and/or comprises mutations that increase the cytosine deaminase's stability compared to a wild-type cytosine deaminase.
31 . The method of claim 19 , wherein 5-FC is administered 6 to 14 days after administering the replicative viral vector to the subject.
32 . The method of claim 19 , wherein one or more doses of 5-FC are orally administered to the subject.
33 . The method of claim 32 , wherein each dose of the one or more doses of 5-FC comprises up to 500 mg/kg of 5-FC.
34 . The method of claim 19 , wherein the cancer is a brain cancer.
35 . The method of claim 34 , wherein the brain cancer is glioblastoma multiforme.
36 . The method of claim 19 , wherein the subject is subjected to radiation when α-fluoro-β-alanine (FBAL) levels are at a therapeutic level that causes increased radiosensitization by tumor cells that convert 5-FC to 5-FU.