Compositions and methods comprising engineered chimeric antigen receptor and modulator of car

Abstract

There is provided method for making a cell composition which comprises step of transducing a population of cells with a mixture of at least two viral vectors, wherein at least one vector comprises a nucleic acid sequence which encodes a chimeric antigen receptor (CAR); and wherein at least one vector comprises a nucleic acid encoding an activity modulator which modulates the activity of the CAR, of a cell expressing the CAR, or of a target cell. There is also provided a cell composition made by such a method and its use in the treatment of diseases such as cancer.

1 . A method for making a cell composition which comprises step of transducing a population of cells with a mixture of at least two viral vectors, wherein at least one vector comprises a nucleic acid sequence which encodes a chimeric antigen receptor (CAR); and wherein at least one vector comprises a nucleic acid encoding an activity modulator which modulates the activity of the CAR, of a cell expressing the CAR, or of a target cell. 
     
     
         2 . A method according to  claim 1 , wherein the activity modulator is a dominant negative SHP-1 or SHP-2. 
     
     
         3 . A method according to  claim 1 , wherein the activity modulator is a dominant negative transforming growth factor (TGF)β receptor. 
     
     
         4 . A method according to  claim 1 , wherein the activity modulator is a constitutively active chimeric cytokine receptor. 
     
     
         5 . A method according to  claim 1 , wherein in the mixture of viral vectors at least one vector comprises a nucleic acid sequence which encodes a dominant negative SHP-1 or SHP-2; and at least one vector comprises a nucleic acid sequence which encodes a dominant negative transforming growth factor (TGF)β receptor. 
     
     
         6 . A method according to  claim 1 , wherein the mixture of viral vectors comprises two, three, four, five or six viral vectors, at least one of which comprises a nucleic acid sequence encoding a CAR; and at least one of which comprises a nucleic acid sequence encoding an activity modulator. 
     
     
         7 . A method for making a cell composition according to  claim 1  which comprises the following steps:
 (i) transducing a population of cells with a mixture of at least two viral vectors; and 
 (ii) selecting CAR-expressing cells from the transduced cell population from step (i). 
 
     
     
         8 . A method according to  claim 1 , wherein each of the viral vectors in the mixture comprises a nucleic acid sequence encoding a CAR. 
     
     
         9 . A viral vector composition which comprises a mixture of at least two viral vectors, wherein at least one vector comprises a nucleic acid sequence which encodes a chimeric antigen receptor (CAR); and wherein at least one vector comprises a nucleic acid encoding an activity modulator which modulates the activity of the CAR, of a cell expressing the CAR, or of a target cell. 
     
     
         10 . A viral vector composition according to  claim 9 , which comprises a first vector and a second vector, both of which comprise a nucleic acid sequence encoding a chimeric antigen receptor (CAR). 
     
     
         11 . A viral vector composition according to  claim 10 , wherein the nucleic acid sequence of the first vector and the nucleic acid sequence of the second vector encode the same CAR. 
     
     
         12 . A viral vector composition according to  claim 10  or  11 , wherein both the first vector and the second vector also comprise a nucleic acid encoding an activity modulator which modulates the activity of the CAR, of a cell expressing the CAR, or of a target cell. 
     
     
         13 . A viral vector composition according to  claim 12 , wherein the activity modulators are selected from: a dominant negative SHP-1 or SHP-2; a dominant negative transforming growth factor (TGF)β receptor; and a constitutively active chimeric cytokine receptor. 
     
     
         14 . A viral vector composition according to  claim 13 , wherein the first vector comprises a nucleic acid sequence encoding a dominant negative SHP-1 or SHP-2 and a nucleic acid sequence encoding a dominant negative transforming growth factor (TGF)β receptor; and the second vector comprises a nucleic acid sequence encoding a constitutively active chimeric cytokine receptor. 
     
     
         15 . A viral vector composition according to  claim 11  wherein the first and second vectors encode the same CAR and the CAR has an antigen-binding domain which binds disialoganglioside (GD2). 
     
     
         16 . A viral vector composition according to  claim 10 , wherein the first vector and/or the second vector comprises a nucleic acid sequence encoding a suicide gene. 
     
     
         17 . A cell composition made by a method according to  claim 1 . 
     
     
         18 . A method for treating a disease in a subject which comprises the step of administering a cell composition according to  claim 17  to the subject. 
     
     
         19 - 20 . (canceled) 
     
     
         21 . A method for determining the optimal combination of components for a CAR-expressing cell to treat a disease, which comprises the following steps:
 (i) administering a cell composition according to  claim 17  to a subject having the disease;   (ii) monitoring the patient or samples from the patient to determine which sub-population of cells in the cell composition show the greatest level of engraftment and/or proliferation; and   (iii) analysing the phenotype/genotype of the cells in the sub-population to ascertain the CAR(s) and/or activity modulator(s) expressed by those cells.