Cancer treatment with recombinant vector

Abstract

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. It further provides use of such modified CDs with a thymosin-alpha-1 polypeptide in the treatment of disease and disorders.

1 . A therapeutic combination comprising a thymosin-1-alpha polypeptide and a replication retroviral vector for use in the treatment of a subject comprising a cell proliferative disease or disorder, wherein the replication competent retroviral vector 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, 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 in a target cell. 
   
     
     
         2 . The combination of  claim 1 , wherein the heterologous polynucleotide comprises a suicide gene that expresses a polypeptide that converts a non-toxic prodrug to a toxic drug. 
     
     
         3 . The combination of  claim 1 , wherein the target cell is a cancer cell. 
     
     
         4 . The combination of  claim 1 , wherein the target cell comprises a cell proliferative disorder. 
     
     
         5 . The combination of  claim 4 , wherein the cell proliferative disorder 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, rheumatoid arthritis or other autoimmune disease. 
     
     
         6 . The combination of  claim 1 , wherein the retroviral vector is administered prior to the thymosin-alpha-1 polypeptide. 
     
     
         7 . The combination of  claim 1 , wherein the retroviral polynucleotide sequence is derived 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). 
     
     
         8 . The combination of  claim 1 , wherein the retroviral envelope is an amphotropic MLV envelope. 
     
     
         9 . The combination of  claim 1 , wherein the retrovirus is a gammaretrovirus. 
     
     
         10 . The combination of  claim 1 , wherein the thymosin-alpha-1 polypeptide comprises at least 85% identity to SEQ ID NO:73 and having thymosin-alpha-1 activity. 
     
     
         11 . The combination of  claim 1 , wherein the heterologous polynucleotide encodes a polypeptide having cytosine deaminase activity. 
     
     
         12 . The combination of  claim 1 , wherein the heterologous polynucleotide is selected from the group consisting of a suicide gene and an immunopotentiating gene. 
     
     
         13 . The combination of  claim 1 , wherein the retrovirus further comprises an miRNA. 
     
     
         14 . The combination of  claim 1 , 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 polynucleotide encoding cytosine deaminase, 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 in a target cell.   
     
     
         15 . The combination of  claim 1 , wherein the thymosin-1-alpha and retroviral vector are formulated for delivery simultaneously. 
     
     
         16 . A method of treating a subject with a cell proliferative disorder comprising:
 administering a thymosin-alpha-1 polypeptide to the subject either before, during or after administration of a replication competent retrovirus, the replication competent retrovirus comprising   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, 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 in a target cell.   
     
     
         17 . The method of  claim 16 , wherein the retroviral polynucleotide sequence is derived 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). 
     
     
         18 . The method of  claim 16 , wherein the retroviral envelope is an amphotropic MLV envelope. 
     
     
         19 . The method of  claim 16 , wherein the retrovirus is a gammaretrovirus. 
     
     
         20 . The method of  claim 16 , wherein the target cell is a neoplastic cell. 
     
     
         21 . The method of  claim 16 , wherein the cell proliferative disorder 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, rheumatoid arthritis or other autoimmune disease. 
     
     
         22 . The method of  claim 16 , wherein the thymosin-alpha-1 polypeptide comprises at least 85% identity to SEQ ID NO:73 and having a thymosin-alpha-1 activity. 
     
     
         23 . The method of  claim 16 , wherein the heterologous polynucleotide encodes a polypeptide having cytosine deaminase activity. 
     
     
         24 . The method of  claim 16 , wherein the heterologous polynucleotide is selected from the group consisting of a suicide gene and an immunopotentiating gene. 
     
     
         25 . The method of  claim 16 , wherein the retrovirus further comprises an miRNA. 
     
     
         26 . The method of  claim 16 , wherein the replication competent retrovirus comprising
 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 polynucleotide encoding cytosine deaminase, 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 in a target cell.