Recombinant vectors
This disclosure provides modified cytosine deaminases (CDs). The disclosure further relates to cells and vector expressing or comprising such modified CDs and methods of using such modified CDs in the treatment of disease and disorders.
1. A recombinant replication competent gammaretrovirus comprising:
a gammaretroviral GAG protein;
a gammaretroviral POL protein;
a gammaretroviral envelope;
a gammaretroviral RNA polynucleotide comprising 3′ untranslated region (U3) and repeat region (R) sequences from murine leukemia virus (MLV), Moloney murine leukemia virus (MoMLV), Feline leukemia virus (FeLV), Baboon endogenous retrovirus (BEV), porcine endogenous virus (PERV), the cat derived retrovirus RD114, squirrel monkey retrovirus, Xenotropic murine leukemia virus-related virus (XMRV), avian reticuloendotheliosis virus (REV), or Gibbon ape leukemia virus (GALV) at the 3′ end of the gammaretroviral polynucleotide sequence, an R and 5′ untranslated region (U5) sequence from MLV, MoMLV, FeLV, BEV, PERV, RD114, squirrel monkey retrovirus, XMRV, REV or GALV at the 5′ end of the gammaretroviral polynucleotide, a gag nucleic acid domain, a pol nucleic acid domain and an env nucleic acid domain from MLV, MoMLV, FeLV, BEV, PERV, RD114, squirrel monkey retrovirus, XMRV, REV or GALV located between the U5 and U3 regions;
a cassette comprising an internal ribosome entry site (IRES) upstream and operably linked to a heterologous polynucleotide encoding an antibody or antibody fragment, wherein the cassette is positioned 5′ to the U3 region and 3′ to the env nucleic acid domain; and
cis-acting sequences necessary for reverse transcription, packaging and integration in a target cell.
2. The retrovirus of claim 1 , wherein the retroviral envelope is an amphotropic MLV envelope.
3. The retrovirus of claim 1 , wherein the retrovirus infects cancer cells of a cancer.
4. The retrovirus of claim 3 , wherein the cancer cell is selected from the group consisting of lung cancer, colon-rectum cancer, breast cancer, prostate cancer, urinary tract cancer, uterine cancer, brain cancer, head and neck cancer, pancreatic cancer, melanoma, stomach cancer and ovarian cancer.
5. The retrovirus of claim 1 , further comprising a promoter cassette comprising a pol III promoter that is positioned 5′ to the U3 region and 3′ to the env nucleic acid domain.
6. The retrovirus of claim 5 , wherein the pol III promoter is an H1 or U6 promoter.
7. The retrovirus according to claim 5 , wherein the promoter cassette comprises the pol III promoter operably linked to a heterologous nucleic acid sequence comprising an inhibitory polynucleotide.
8. The retrovirus according to claim 7 , wherein the inhibitory polynucleotide comprises an miRNA, RNAi or siRNA sequence.
9. A method of treating a cell proliferative disorder comprising administering the recombinant replication competent gammaretrovirus of claim 1 to a subject having a cell proliferative disorder under conditions such that the retrovirus infects cells with the disorder and contacting the subject with an anti-cancer agent or chemotherapeutic agent, wherein the retrovirus is administered from about 10 3 to 10 11 TU/g brain weight.
10. The method of claim 9 , wherein the anti-cancer agent is selected from the group consisting of bevacizumab, pegaptanib, ranibizumab, sorafenib, sunitinib, AE-941, VEGF Trap, pazopanib, vandetanib, vatalanib, cediranib, fenretinide, squalamine, INGN-241, oral tetrathiomolybdate, tetrathiomolybdate, Panzem NCD, 2-methoxyestradiol, AEE-788, AG-013958, bevasiranib sodium, AMG-706, axitinib, BIBF-1120, CDP-791, CP-547632, PI-88, SU-14813, SU-6668, XL-647, XL-999, IMC-1121B, ABT-869, BAY-57-9352, BAY-73-4506, BMS-582664, CEP-7055, CHIR-265, CT-322, CX-3542, E-7080, ENMD-1198, OSI-930, PTC-299, Sirna-027, TKI-258, Veglin, XL-184, or ZK-304709.