Engineered Dendritic Cells and Uses for the Treatment of Cancer

Abstract

This invention provides the field of therapeutics. Most specifically present invention provides methods of generating in vitro engineered dendritic cells conditionally expressing interleukin-12 (IL-12) under the control of a gene expression modulation system in the presence of activating ligand and uses for therapeutic purposes in animals including human.

1 . A vector for conditionally expressing a protein having the function of interleukin-12 (IL 12), comprising a polynucleotide encoding a gene switch, said gene switch comprising (1) at least one transcription factor sequence, wherein said at least one transcription factor sequence encodes a ligand-dependent transcription factor, operably linked to a promoter, and (2) a polynucleotide encoding a protein having the function of IL-12 linked to a promoter which is activated by said ligand-dependent transcription factor. 
     
     
         2 . The vector of  claim 1 , wherein said polynucleotide encoding a gene switch comprises (1) at least one transcription factor sequence, operably linked to a promoter, wherein said at least one transcription factor sequence encodes a ligand-dependent transcription factor, and (2) a polynucleotide encoding a protein having the function of IL-12 linked to a promoter which is activated by said ligand-dependent transcription factor. 
     
     
         3 . The vector of  claim 1 , which is an adenoviral vector. 
     
     
         4 . (canceled) 
     
     
         5 . The vector of  claim 1 , wherein said gene switch is an ecdysone receptor (EcR)-based gene switch. 
     
     
         6 . The vector of  claim 1 , wherein said polynucleotide encoding a gene switch comprises a first transcription factor sequence and a second transcription factor sequence under the control of a promoter, wherein the proteins encoded by said first transcription factor sequence and said second transcription factor sequence interact to form a protein complex which functions as a ligand-dependent transcription factor. 
     
     
         7 . The vector of  claim 6 , wherein said first transcription factor and said second transcription factor are connected by an internal ribosomal entry site. 
     
     
         8 . (canceled) 
     
     
         9 . The vector of  claim 1 , wherein said polynucleotide encoding a gene switch comprises a first transcription factor sequence under the control of a first promoter and a second transcription factor sequence under the control of a second promoter, wherein the proteins encoded by said first transcription factor sequence and said second transcription factor sequence interact to form a protein complex which functions as a ligand-dependent transcription factor. 
     
     
         10 . (canceled) 
     
     
         11 . (canceled) 
     
     
         12 . The vector of  claim 1 , wherein said vector further comprises a polynucleotide encoding a protein having the function of IFN-alpha linked to a promoter which is activated by said ligand-dependent transcription factor. 
     
     
         13 . (canceled) 
     
     
         14 . The vector of  claim 1 , wherein said vector comprises a polynucleotide encoding a protein having the function of IL-12 wherein said protein comprises a polypeptide at least 85% identical to wild type human IL-12. 
     
     
         15 . A method of producing a population of dendritic cells conditionally expressing a protein having the function of IL-12, comprising modifying at least a portion of dendritic cells by introducing into said dendritic cells the vector of  claim 1 . 
     
     
         16 . (canceled) 
     
     
         17 . (canceled) 
     
     
         18 . An in vitro engineered dendritic cell comprising the vector of  claim 1 . 
     
     
         19 . The in vitro engineered dendritic cell of  claim 18  further comprising a vector conditionally expressing a protein having the function of IFN-alpha, said vector comprises a polynucleotide encoding a gene switch, wherein said polynucleotide comprises (1) at least one transcription factor sequence which is operably linked to a promoter, wherein said at least one transcription factor sequence encodes a ligand-dependent transcription factor, and (2) a polynucleotide encoding a protein having the function of IFN-alpha linked to a promoter which is activated by said ligand-dependent transcription factor. 
     
     
         20 . A pharmaceutical composition comprising a population of the in vitro engineered dendritic cells of  claim 18 . 
     
     
         21 - 28 . (canceled) 
     
     
         29 . Use of a population of the in vitro engineered cells of  claim 18 , wherein said cells are to be administered to a mammal in need thereof. 
     
     
         30 - 32 . (canceled) 
     
     
         33 . The use according to  claim 29 , further comprising administration of a ligand, wherein said ligand is administered intratumorally, orally, intraperitoneally, or subcutaneously. 
     
     
         34 . (canceled) 
     
     
         35 . (canceled) 
     
     
         36 . The use of  claim 33 , wherein said ligand is administered less than 48 hours after said in vitro engineered dendritic cell. 
     
     
         37 . (canceled) 
     
     
         38 . (canceled) 
     
     
         39 . A method for determining the efficacy of an in vitro engineered dendritic cell based therapeutic regime in a patient comprising:
 (a) measuring the level of expression or the level of activity or both of interferon-gamma (IFN-γ) in a first biological sample obtained from said patient in need thereof before administration of in vitro engineered dendritic cells, thereby generating a control level;   (b) administering to a patient in need thereof, the in vitro engineered dendritic cell of  claim 18 ;   (c) administering to said patient in need thereof an effective amount of an activating ligand;   (d) measuring the level of expression or the level of activity or both of IFN-γ in a second biological sample obtained from said patient in need thereof following administration of in vitro engineered DC and activating ligand, thereby generating a test level; and   (e) comparing the control level to the test level of IFN-γ, wherein an increase in the test level of expression, activity or both of IFN-γ relative to the control level indicates that the therapeutic regime is effective in said patient in need thereof.   
     
     
         40 - 50 . (canceled) 
     
     
         51 . A kit comprising said in vitro engineered dendritic cells of  claim 18 . 
     
     
         52 . (canceled) 
     
     
         53 . Use of a population of the in vitro engineered dendritic cells of  claim 18  in the manufacture of a medicament for inducing conditional expression of a protein having the function of interleukin-12 (IL-12) in the dendritic cells. 
     
     
         54 - 73 . (canceled)