T cell which expresses a gamma-delta t cell receptor (tcr) and a chimeric antigen receptor (car)

Abstract

The present invention provides a T cell which expresses a gamma-delta T cell receptor (TCR) and a chimeric antigen receptor (CAR), wherein the CAR comprises: an antigen binding domain; a transmembrane domain; and a co-stimulatory intracellular signalling domain; wherein the intracellular signalling domain provides a co-stimulatory signal to the T cell following binding of antigen to the antigen binding domain.

1 . A T cell which expresses a gamma-delta T cell receptor (TCR) and a chimeric antigen receptor (CAR), wherein the CAR comprises;
 (i) an antigen binding domain;   (ii) a transmembrane domain; and   (iii) a co-stimulatory intracellular signalling domain;   
       wherein the intracellular signalling domain provides a co-stimulatory signal to the T cell following binding of antigen to the antigen binding domain. 
     
     
         2 . A cell according to  claim 1  wherein the antigen binding domain is capable of binding to a tumour-associated antigen (TAA). 
     
     
         3 . A cell according to  claim 1  wherein the antigen binding domain is capable of binding to GD2, CD33, CD19 or EGFR. 
     
     
         4 . A cell according to any preceding claim wherein the transmembrane domain comprises a CD8 stalk or a CD28 transmembrane domain. 
     
     
         5 . A cell according to any of  claims 1  to  4  wherein the intracellular signalling domain comprises the DAP10, CD28, CD27, 41BB, OX40, CD30, IL2-R, IL7-R, IL21-R, NKp30, NKp44 or DNAM-1 (CD226) signalling domain. 
     
     
         6 . A cell according to any of  claims 1  to  4  wherein the intracellular signalling domain comprises the DAP10 signalling domain. 
     
     
         7 . A cell according to any preceding claim wherein the binding of a first antigen to the gamma-delta TCR results in signal 1 production and binding of a second antigen to the antigen binding domain of the CAR results in signal 2 production. 
     
     
         8 . A cell according to any preceding claim wherein the CAR further comprises a spacer domain between the antigen binding domain and the transmembrane domain, for example a CD8 stalk or an Fc region. 
     
     
         9 . A cell according to any preceding claim wherein the gamma-delta TCR is capable of binding to a phosphoantigen; major histocompatibility complex class I chain-related A (MICA); major histocompatibility complex class I chain-related B (MICB); NKG2D ligand 1-6 (ULBP 1-6); CD1c; CD1d; endothelial protein C receptor (EPCR); lipohexapeptide; phycoreythrin or histidyl-tRNA-synthase. 
     
     
         10 . A CAR comprising;
 (i) an antigen-binding domain;   (ii) a transmembrane domain; and   (iii) an intracellular signalling domain;   
       wherein the intracellular signalling domain comprises a co-stimulatory intracellular signalling domain but does not comprise a CD3 endodomain. 
     
     
         11 . A CAR according to  claim 10  wherein the co-stimulatory intracellular signalling domain is selected from a DAP10, CD28, CD27, 41BB, OX40, CD30, IL2-R, IL7-R, IL21-R, NKp30, NKp44 or DNAM-1 (CD226) signalling domain. 
     
     
         12 . A CAR comprising;
 (i) an antigen-binding domain;   (ii) a transmembrane domain; and   (iii) an intracellular signalling domain;   
       wherein the intracellular signalling domain comprises a DAP10 signalling domain. 
     
     
         13 . A CAR according to  claim 12  wherein the intracellular signalling domain does not comprise a CD3 endodomain. 
     
     
         14 . A CAR according to any of  claims 10  to  13  which is a CAR as defined in any of  claims 2  to  9 . 
     
     
         15 . A nucleic acid sequence encoding a CAR as defined in any preceding claim. 
     
     
         16 . A vector comprising a nucleic acid sequence as defined in  claim 15 . 
     
     
         17 . A vector according to  claim 16  which is a retroviral vector, a lentiviral vector or a transposon. 
     
     
         18 . A method for making a cell according to any of  claims 1  to  9 , which comprises the step of introducing: a nucleic acid sequence according to  claim 15  or a vector according to  claim 16  or  17  into a cell. 
     
     
         19 . A method according to  claim 18  wherein the cell is stimulated with a gamma delta T cell stimulating agent. 
     
     
         20 . A method according to  claim 19  wherein the gamma-delta T cell stimulating agent is selected from isopentenyl pyrophosphate (IPP); analogs of IPP; and inhibitors of farnesyl pyrophosphate synthase (FPPS). 
     
     
         21 . A method according to  claim 19  or  20 , wherein the cell is from a sample isolated from a subject. 
     
     
         22 . A pharmaceutical composition comprising a cell according to any of  claims 1  to  9 , a CAR according to any of  claims 10  to  14 , a nucleic acid sequence according to  claim 15  or a vector according to  claim 16  or  17 . 
     
     
         23 . A method for treating a disease, which comprises the step of administering a pharmaceutical composition according to  claim 22  to a subject. 
     
     
         24 . A method according to  claim 23  which comprises the step of administering a gamma-delta T cell stimulating agent to the subject. 
     
     
         25 . A method according to  claim 24  wherein the gamma-delta T cell stimulating agent is selected from isopentenyl pyrophosphate (IPP); analogs of IPP; and inhibitors of farnesyl pyrophosphate synthase (FPPS). 
     
     
         26 . A method according to any of  claims 23  to  25 , which comprises the following steps:
 (i) isolation of a cell-containing sample from a subject; 
 (ii) transduction or transfection of cells with: a nucleic acid according to  claim 15  or a vector according to  claim 16  or  17 ; and 
 (iii) administering the cells from (ii) to the subject. 
 
     
     
         27 . A pharmaceutical composition according to  claim 22  for use in treating and/or preventing a disease. 
     
     
         28 . The use of a cell according to any of  claims 1  to  9  in the manufacture of a medicament for treating and/or preventing a disease. 
     
     
         29 . A method according to any of  claims 23  to  26  or a use according to  claim 24  or  25  wherein the disease is cancer, microbial infection or viral infection. 
     
     
         30 . A method according to any of  claims 23  to  26  or a use according to  claim 27  or  28  wherein the disease is cancer.