Adenovectors for delivery of therapeutic genetic material into t cells

Abstract

The invention provides adenoviral vectors and compositions for the highly efficient transduction of T cells.

1 . (canceled) 
     
     
         2 . A serotype 28 adenoviral vector comprising (i) a fiber protein containing a high affinity RGD ligand and (ii) an exogenous nucleic acid sequence. 
     
     
         3 . The vector of  claim 1 , which requires complementation of a deficiency in one or more early regions of the adenoviral genome for propagation and does not require complementation of any other deficiency of the adenoviral genome for propagation. 
     
     
         4 .- 8 . (canceled) 
     
     
         9 . The vector of  claim 2 , wherein the vector comprises an RGD or pK7 modification to the knob. 
     
     
         10 . The vector of  claim 2 , wherein the exogenous nucleic acid sequence encodes single chain variable anti-PSMA, single chain variable anti-PSMA, single chain variable anti-MUC1, single chain variable anti-CD19, single chain variable anti-ROR1, single chain variable anti-mesothelin, single chain variable anti-CD2, single chain variable anti-CD123, single chain variable anti-MUC16, single chain variable anti-Her2/Neu, single chain variable anti-L1CAM, or single chain variable anti-BCMA. 
     
     
         11 . The vector of  claim 2 , wherein the exogenous nucleic acid sequence encodes an αβ or γδ T cell receptor (TCR). 
     
     
         12 . The vector of  claim 11 , wherein the TCR is specific for a cancer or infectious disease epitope. 
     
     
         13 . (canceled) 
     
     
         14 . The vector of  claim 2 , wherein the exogenous nucleic acid sequence encodes a DNA endonuclease. 
     
     
         15 . The vector of  claim 14 , wherein the DNA endonuclease is selected from the group consisting of CRISPR/Cas9, meganuclease, TALENS, zinc finger nuclease (ZFN), and Cpf1. 
     
     
         16 . The vector of  claim 2 , wherein the exogenous nucleic acid sequence is transcribed to produce a guide RNA sequence or siRNA sequence. 
     
     
         17 . (canceled) 
     
     
         18 . The vector of  claim 2 , wherein the exogenous nucleic acid sequence encodes rimiducid inducible caspase-9 or herpes simplex virus-thymidine kinase. 
     
     
         19 . The vector of  claim 2 , wherein vector comprises a promoter operably linked to the exogenous nucleic acid sequence. 
     
     
         20 . The vector of  claim 19 , wherein the promoter is selected from the group consisting of the CD3, CD4, CD8, EF-1, PGK, RSV, Beta-Actin, CMV, MCK, UB, and HIV LTR promoter. 
     
     
         21 . A cell comprising
 (a) the vector of  claim 2 ,   (b) a serotype 28 adenoviral vector comprising (i) at least a portion of an adenovirus serotype 45 hexon protein in place of at least a portion of a corresponding endogenous serotype 28 hexon protein and (ii) an exogenous nucleic acid sequence;   (c) a serotype 28 adenoviral vector comprising (i) at least a portion of an adenovirus serotype 45 fiber protein in place of at least a portion of a corresponding endogenous serotype 28 fiber protein and (ii) an exogenous nucleic acid sequence,   (d) a serotype 28 adenoviral vector comprising (i) at least a portion of an adenovirus serotype 45 hexon protein in place of at least a portion of a corresponding endogenous serotype 28 hexon protein, (ii) at least a portion of an adenovirus serotype 45 fiber protein in place of at least a portion of a corresponding endogenous serotype 28 fiber protein, and (iii) an exogenous nucleic acid sequence, or   (e) a serotype 35 adenoviral vector comprising an exogenous nucleic acid sequence.   
     
     
         22 . The cell of  claim 21 , which is a T cell. 
     
     
         23 . (canceled) 
     
     
         24 . A composition comprising (i) the adenoviral vector of  claim 2  and (ii) a pharmaceutically acceptable carrier. 
     
     
         25 . A method of transducing T cells comprising contacting the T cells with
 (a) the vector of  claim 2 ,   (b) a serotype 28 adenoviral vector comprising (i) at least a portion of an adenovirus serotype 45 hexon protein in place of at least a portion of a corresponding endogenous serotype 28 hexon protein and (ii) an exogenous nucleic acid sequence;   (c) a serotype 28 adenoviral vector comprising (i) at least a portion of an adenovirus serotype 45 fiber protein in place of at least a portion of a corresponding endogenous serotype 28 fiber protein and (ii) an exogenous nucleic acid sequence,   (d) a serotype 28 adenoviral vector comprising (i) at least a portion of an adenovirus serotype 45 hexon protein in place of at least a portion of a corresponding endogenous serotype 28 hexon protein, (ii) at least a portion of an adenovirus serotype 45 fiber protein in place of at least a portion of a corresponding endogenous serotype 28 fiber protein, and (iii) an exogenous nucleic acid sequence, or   (e) a serotype 35 adenoviral vector comprising an exogenous nucleic acid sequence, thereby transducing T cells with the vector.   
     
     
         26 . The method of  claim 25 , wherein the T cells are activated prior to contacting the T cells with the vector. 
     
     
         27 . The method of  claim 26 , wherein the T cells are activated by contacting the T cells with anti-CD3 antibodies and a costimulatory molecule. 
     
     
         28 . The method of  claim 27 , wherein the costimulatory molecule is selected from the group consisting of CD28, CD137/4-1BB, CD40, CD40L, ICOS, OX40, CD2, LFA1, galectin 9, GITR or a combination thereof. 
     
     
         29 . The method of  claim 26 , wherein the activated T cells are cultured in serum-free medium prior to contact with the vector. 
     
     
         30 . The method of  claim 29 , wherein the serum-free medium comprises a cytokine selected from the group consisting of IL-2, IL-7, IL-12, IL-15, IL-21, or a combination thereof. 
     
     
         31 . The method of  claim 25 , wherein the T cells are from a patient with cancer or an infection. 
     
     
         32 . The method of  claim 25 , wherein the T cells are contacted with the vector 1-15 days after the T cells are activated.