Vectors conditionally expressing therapeutic proteins, host cells comprising the vectors, and uses thereof

Abstract

This invention relates to the field of therapeutics. Most specifically, the invention provides methods of generating conditionally expressing vectors for one or more immunomodulators under the control of a gene expression modulation system in the presence of activating ligand and uses for therapeutic purposes in animals. These vector may be provided to treat a variety of disorders, e.g., neoplastic disorders, through direct injection or through in vitro engineered cells, such as dendritic cells.

1 . A vector encoding one or more therapeutic proteins, wherein said vector comprises a first polynucleotide encoding a gene switch, wherein said first polynucleotide comprises (1) at least one transcription factor sequence which is operably linked to a first promoter, wherein said at least one transcription factor sequence encodes a ligand-dependent transcription factor, and (2) a second polynucleotide encoding said one or more therapeutic proteins, wherein said second polynucleotide is operably linked to a second promoter which is activated by said ligand-dependent transcription factor. 
     
     
         2 . The vector of  claim 1 , wherein the one or more therapeutic proteins is selected from the group consisting of erythropoetin, ghrelin, osteoprotegerin, RANKL, RANKL decoy, TNF-α antagonist, an IL-1 antagonist, G-CSF, GM-CSF, IFN-α, angiostatin, endostatin, TNF-α, PP1DCY-LSRLOC, β-glucuronidase, IL-12, a-galactosidase A, Arylsulfatase A, a-glucosidase, b-glucosidase, glucocerebrosidase, CLN6 protein, Juvenile associated with CLN3, N-sulfoglucosamine sulfohyrolase (SGSH), a-N-acetylglucosaminidase, acetyl-CoA-glucosaminide acetyltransferase, N-acetylglucosamine-6-sulfatase, a-L-iduronidase, arylsulfatase B, acid sphingomyelinase, iuduronate sulfatase, and ceruloplasmin. 
     
     
         3 . The vector of  claim 1 , wherein the one or more therapeutic proteins is selected from the group consisting of IL-1, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10R DN or a subunit thereof, IL-15, IL-18, IL-21, IL-23, IL-24, IL-27, GM-CSF, IFN-alpha, IFN-gamma, IFN-alpha 1, IFN alpha 2, IL-15-R-alpha, CCL3 (MIP-1a), CCL5 (RANTES), CCL7 (MCP3), XCL1 (lymphotactin), CXCL1 (MGSA-alpha), CCR7, CCL19 (MIP-3b), CXCL9 (MIG), CXCL10 (IP-10), CXCL12 (SDF-1), CCL21 (6Ckine), OX40L, 4-1BBL, CD40, CD70, GITRL, LIGHT, b-Defensin, HMGB1, Flt3L, IFN-beta, TNF-alpha, dnFADD, BCG, TGF-alpha, PD-L1 RNAi, a PD-L1 antisense oligonucleotide, TGFbRII DN, ICOS-L, 5100, CD40L, p53, survivin, p53-survivin fusion, MAGE3, PSA and PSMA. 
     
     
         4 . The vector of  claim 1 , wherein said vector is selected from the group consisting of plasmid, adenovirus, retrovirus, adeno-associated virus, pox virus, baculovirus, vaccinia virus, herpes simplex virus, Epstein-Barr virus, adenovirus, geminivirus, caulimovirus, liposomes, electrically charged lipids (cytofectins), DNA-protein complexes, and biopolymers. 
     
     
         5 . (canceled) 
     
     
         6 . The vector of  claim 1 , further comprising a polynucleotide encoding IL-12. 
     
     
         7 . (canceled) 
     
     
         8 . The vector of  claim 1 , wherein said gene switch is an ecdysone receptor (EcR)-based gene switch. 
     
     
         9 - 10 . (canceled) 
     
     
         11 . The vector of  claim 1 , wherein said first transcription factor sequence and said second transcription factor sequence are connected by an EMCV internal ribosomal entry site (IRES). 
     
     
         12 . The vector of  claim 1 , wherein said second polynucleotide encoding the one or more therapeutic proteins encodes human proteins. 
     
     
         13 . The vector of  claim 6 , wherein said polynucleotide encoding IL-12 encodes human IL-12. 
     
     
         14 - 31 . (canceled) 
     
     
         32 . A population of immune cells or therapy support cells (TSC) expressing one or more therapeutic proteins, comprising the vector of  claim 1 . 
     
     
         33 - 38 . (canceled) 
     
     
         39 . A pharmaceutical composition comprising the vector of  claim 4 . 
     
     
         40 - 63 . (canceled) 
     
     
         64 . The vector of  claim 1 , wherein said ligand-dependent transcription factor is activated by a diacylhydrazine ligand. 
     
     
         65 . The vector of  claim 64 , wherein said diacylhydrazine ligand is selected from RG-115819, RG-115932, and RG-115830. 
     
     
         66 - 73 . (canceled) 
     
     
         74 . Use of a vector in the manufacturing of a medicament for treating a disease or disorder in a mammal,
 wherein said vector comprises a first polynucleotide encoding a gene switch, wherein said gene switch comprises   
       at least one transcription factor sequence which is operably linked to a first promoter, wherein said at least one transcription factor sequence encodes a ligand-dependent transcription factor, and 
       wherein said vector comprises a second polynucleotide encoding one or more proteins operably linked to a second promoter which is activated by said ligand-dependent transcription factor, 
       wherein said vector is not contained within a cell. 
     
     
         75 . The use of  claim 74 , wherein the disease is selected from the group consisting of chronic renal disease, osteoarthritis, oncology, viral upper respiratory infection, feline plasma cell stomatitis, feline eosinophillic granulomas, feline leukemia virus infection, canine distemper infection, systemic fungal infections, cardiomyopathy, mucopolysaccharidosis VII, and infectious disease. 
     
     
         76 - 84 . (canceled) 
     
     
         85 . Use of a vector in the manufacturing of a medicament for treating a liver disease in a mammal,
 wherein said vector comprises a first polynucleotide encoding a gene switch, wherein said gene switch comprises   
       at least one transcription factor sequence which is operably linked to a first promoter, wherein said at least one transcription factor sequence encodes a ligand-dependent transcription factor, and 
       wherein said vector comprises a second polynucleotide encoding one or more proteins operably linked to a second promoter which is activated by said ligand-dependent transcription factor. 
     
     
         86 . The use of  claim 85 , wherein the liver disease is Hepatitis B. 
     
     
         87 . The use of  claim 85 , wherein the liver disease is Hepatitis C. 
     
     
         88 . The use of  claim 85 , wherein one of said one or more proteins is IFN-α. 
     
     
         89 . The use of  claim 85 , wherein one of said one or more proteins is ceruloplasmin. 
     
     
         90 - 102 . (canceled)