COMPOSITIONS AND METHODS COMPRISING ENGINEERED CHIMERIC ANTIGEN RECEPTOR AND MODULATOR OF CAR
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.