Modified gamma delta t cells and uses thereof

Abstract

The present invention provides composition and methods for the treatment of cancer or infectious diseases in a human. The invention includes the generation and administration of gamma delta T cells that express chimeric antigen receptors (CARs) comprising an antigen binding domain, a hinge domain, a transmembrane domain, a costimulatory signalling domain with the inclusion or not of a CD3 zeta signalling domain. Expression of CAR sequence omitting the CD3 zeta signalling domain in gamma delta T cells, provides for a CAR-T therapy in vivo, which will effect cytolysis only on target cells providing ligands for activation of the gamma delta T cell receptor (TCR).

1 - 54 . (canceled) 
     
     
         55 . A method of treatment comprising; administering a therapeutically effective amount of modified γδT cells to a recipient subject in need thereof, wherein the modified γδT cells comprise a chimeric antigen receptor (CAR), wherein the CAR comprises (i) an extracellular antigen binding domain having binding specificity to a disease antigen, (ii) a hinge region; (iii) a transmembrane domain, and (iv) one or more co-stimulatory signalling regions capable of providing a co-stimulatory signal, and wherein the CAR does not provide a TCR cell receptor signal. 
     
     
         56 . The method of claim  1 , wherein the modified γδT cells comprise γδT cells modified to express the CAR. 
     
     
         57 . The method of  claim 56 , wherein the γδT cells are collected from a donor subject. 
     
     
         58 . The method of  claim 55 , wherein the modified γδT cells are of a Vγ9Vδ2 isotype. 
     
     
         59 . The method of  claim 55 , wherein the modified γδT cells are activated by a phosphoantigen or by isopentenyl pyrophosphate (IPP). 
     
     
         60 . The method of  claim 55 , wherein the modified γδT cells further comprise an inhibitory chimeric antigen receptor (ICAR), wherein the binding of an antigen by the ICAR inhibits the co-stimulatory signal. 
     
     
         61 . The method of  claim 55 , wherein the extracellular antigen binding domain is capable of binding to: (a) a cell surface target or natural ligand found in or associated with a cell infection, a viral infection, a bacterial infection, a fungal infection, or protozoan infection; (b) an active or inactivated viral fragment, peptide, protein or antigenic segment from a virus; or (c) a tumor-specific antigen or tumor-associated antigen. 
     
     
         62 . The method of  claim 57 , wherein the recipient subject is a different subject to the donor subject. 
     
     
         63 . The method of  claim 62 , wherein the recipient subject is allogeneic to the donor subject. 
     
     
         64 . The method of  claim 57 , wherein the recipient subject is the donor subject. 
     
     
         65 . The method of  claim 55 , wherein the subject has cancer. 
     
     
         66 . The method of  claim 55 , wherein the subject has a viral infection, a bacterial infection, a fungal infection, or a protozoan infection. 
     
     
         67 . The method of  claim 55 , wherein the modified γδT cells are co-administered to the subject with an agent capable of enhancing the levels of phosphoantigens present on target cells via dysregulation of the physiological mevalonate pathway. 
     
     
         68 . The method of  claim 67 , wherein the agent is an aminobisphosphonate. 
     
     
         69 . A method of preparing a modified γδT cell comprising: modifying one or more of γδT cells to incorporate a chimeric antigen receptor (CAR) wherein the CAR comprises (i) an extracellular antigen binding domain having binding specificity to a disease antigen, (ii) an hinge region, (iii) a transmembrane domain, and (iv) one or more co-stimulatory signalling regions capable of providing a co-stimulatory signal, wherein the CAR does not provide a TCR cell receptor signal. 
     
     
         70 . The method of  claim 69 , further comprising, prior to said modifying, collecting the one or more γδT cells from a donor subject. 
     
     
         71 . The method of  claim 69 , wherein modifying the one or more γδT cells comprises incorporating or transducing the one or more γδT cells with a nucleic acid encoding a chimeric antigen receptor (CAR), wherein the CAR comprises an extracellular antigen recognition domain, a hinge, a transmembrane domain, and one or more co-stimulatory signalling regions wherein the CAR does not provide a TCR cell receptor signal, and wherein nucleic acid encodes a polypeptide comprising the amino acid sequences ofSEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and SEQ ID NO: 11. 
     
     
         72 . The method of  claim 71 , wherein the nucleic acid comprises the nucleotide sequence of SEQ ID NO: 7. 
     
     
         73 . A method of treatment comprising; administering to a subject in need thereof a modified γδT cell, wherein the modified γδT cell comprises:
 (i) a γδT cell receptor, wherein the γδT cell receptor provides a T cell receptor (TCR) signal upon binding; and 
 (ii) at least one co-stimulatory chimeric antigen receptor (CAR) wherein the co-stimulatory chimeric antigen receptor is a synthetic receptor to recognise and target a cell surface target, connected to a transmembrane domain which traverses the cell membrane and connects to an intracellular co-stimulatory signaling region capable of providing a co-stimulatory signal, wherein the at least one co-stimulatory CAR provides the costimulatory signal upon binding the cell surface target, and wherein the at least one co-stimulatory CAR does not provide a TCR cell receptor signal. 
 
     
     
         74 . The method of treatment of  claim 73  wherein the modified γδT cell, further comprises an inhibitory chimeric antigen receptor (ICAR), wherein the binding of an antigen by the ICAR inhibits the co-stimulatory signal. 
     
     
         75 . The method of treatment of  claim 73  wherein the modified γδT cell comprises at least one co-stimulatory chimeric antigen receptor (CAR) capable of binding to a cell surface target or natural ligand found in or associated with cell infection, bacterial infection, fungal infection or protozoan infection; an active or inactivated viral fragment; a peptide; a protein; an antigenic segment from a virus; a tumour-specific antigen or tumour associated antigen.