Chimeric antigen receptor dendritic cell (car-dc) for treatment of cancer

Abstract

The current invention provides monocytic cells transfected with chimeric antigen receptor (CAR) to selectively home to tumors and upon homing differentiate into dendritic cells capable of activating immunity which is inhibitory to said tumor. In one embodiment of the invention, monocytic cells are transfected with a construct encoding an antigen binding domain, a transcellular or structural domain, and an intracellular signaling domain. In one specific aspect of the invention, the antigen binding domain interacts with sufficient affinity to a tumor antigen, capable of triggering said intracellular domain to induce an activation signal to induce monocyte differentiation into DC.

1 .- 20 . (canceled) 
     
     
         21 . A pharmaceutical composition comprising:
 (a) a population of modified human monocytes isolated from a blood sample from a human cancer patient by isolating cells expressing CD14, wherein the modified human monocytes comprise a chimeric antigen receptor (CAR), and   (b) a pharmaceutically acceptable excipient.   
     
     
         22 . The pharmaceutical composition of  claim 21 , wherein the CAR comprises an antigen binding domain, a transmembrane domain and an intracellular domain of a stimulatory and/or co-stimulatory molecule. 
     
     
         23 . The pharmaceutical composition of  claim 21 , wherein the antigen binding domain comprises an anti-HER2 antigen binding domain. 
     
     
         24 . The pharmaceutical composition of  claim 21 , wherein the antigen binding domain of the CAR comprises an antibody selected from the group consisting of a chimeric antibody, synthetic antibody, humanized antibody, single heavy chain antibody, single light chain antibody, scFv, and antigen-binding fragments thereof. 
     
     
         25 . The pharmaceutical composition of  claim 21 , wherein the transmembrane domain of the CAR comprises a CD8 transmembrane domain. 
     
     
         26 . The pharmaceutical composition of  claim 21 , wherein the intracellular domain of the CAR comprises CD80 and/or CD86 and/or CD40L and/or OX40L signaling domains. 
     
     
         27 . The pharmaceutical composition of  claim 23 , wherein the intracellular domain of the CAR comprises a CD3 zeta intracellular domain. 
     
     
         28 . The pharmaceutical composition of  claim 21 , wherein the population of modified human monocytes are isolated from a blood sample from a human cancer patient by isolating cells expressing CD14 by magnetic activated cell sorting (MACS) or fluorescent activated cell sorting (FACS). 
     
     
         29 . The pharmaceutical composition of  claim 21 , wherein the modified human monocytes exhibit targeted effector activity. 
     
     
         30 . The pharmaceutical composition of  claim 29 , wherein the targeted effector activity is directed against a target cell comprising an antigen that specifically binds the antigen binding domain of the CAR. 
     
     
         31 . The pharmaceutical composition of  claim 29 , wherein the targeted effector activity is selected from the group consisting of phagocytosis, cytotoxicity, and antigen presentation. 
     
     
         32 . The pharmaceutical composition of  claim 29 , wherein the targeted effector activity is enhanced by inhibition of CD47 or signal-regulatory protein a activity. 
     
     
         33 . The pharmaceutical composition of  claim 21 , further comprising an agent, wherein the agent is selected from the group consisting of a nucleic acid, a peptide and any combination thereof. 
     
     
         34 . The pharmaceutical composition of  claim 21 , wherein >75% of the modified cells in the pharmaceutical composition express the CAR or wherein the modified human monocytes in the population of cells are genetically modified to stably express the CAR. 
     
     
         35 . A method for stimulating an immune response to a target tumor cell or tumor tissue in a subject comprising administering to a subject a therapeutically effective amount of the pharmaceutical composition of  claim 21 . 
     
     
         36 . A method of modifying a population of human cells, comprising: isolating monocytes from a blood sample from a human cancer patient and introducing a chimeric antigen receptor (CAR) into the monocytes isolated from the blood sample, wherein isolating monocytes comprises isolating cells expressing CD14. 
     
     
         37 . The method of  claim 36 , wherein introducing the CAR into the monocytes isolated from the blood sample comprises introducing a nucleic acid sequence encoding the CAR. 
     
     
         38 . The method of  claim 37 , wherein introducing the nucleic acid sequence comprises introducing an mRNA encoding the CAR into the monocytes isolated from the blood sample. 
     
     
         39 . The method of  claim 37 , wherein introducing the nucleic acid sequence comprises transducing the monocytes isolated from the blood sample with a viral vector comprising a nucleic acid sequence encoding the CAR. 
     
     
         40 . The method of  claim 36 , wherein the CAR comprises an antigen binding domain, a transmembrane domain and an intracellular domain of a stimulatory and/or co-stimulatory molecule. 
     
     
         41 . The method of  claim 40 , wherein the antigen binding domain comprises an anti-HER2 antigen binding domain. 
     
     
         42 . The method of  claim 40 , wherein the transmembrane domain of the CAR comprises a CD8 transmembrane domain. 
     
     
         43 . The method of  claim 40 , wherein the intracellular domain of the CAR comprises CD80 and/or CD86 and/or CD40L and/or OX40L signaling domains. 
     
     
         44 . The method of  claim 40 , wherein the intracellular domain of the CAR comprises a CD3 zeta intracellular domain. 
     
     
         45 . The method of  claim 36 , wherein isolating monocytes comprises isolating cells expressing CD14 by magnetic activated cell sorting (MACS) or fluorescent activated cell sorting (FACS). 
     
     
         46 . The method of  claim 36 , wherein the monocytes isolated from the blood sample into which the CAR was introduced exhibit targeted effector activity. 
     
     
         47 . The method of  claim 46 , wherein the targeted effector activity is directed against a target cell comprising an antigen that specifically binds the antigen binding domain of the CAR. 
     
     
         48 . The method of  claim 46 , wherein the targeted effector activity is selected from the group consisting of phagocytosis, cytotoxicity, and antigen presentation. 
     
     
         49 . A modified M1 macrophage or monocyte comprising a chimeric antigen receptor (CAR), wherein the CAR comprises an antigen binding domain, a transmembrane domain and an intracellular domain of a stimulatory and/or co-stimulatory molecule, and wherein the modified M1 macrophage or monocyte comprises an upregulated M1 marker. 
     
     
         50 . The modified M1 macrophage or monocyte of  claim 49 , wherein the upregulated M1 marker is HLA DR. 
     
     
         51 . The modified M1 macrophage or monocyte of  claim 49 , wherein the M1 macrophage or monocyte is not an M2 macrophage or monocyte. 
     
     
         52 . The modified M1 macrophage or monocyte of  claim 49 , wherein the antigen binding domain of the CAR comprises an antibody selected from the group consisting of a chimeric antibody, synthetic antibody, humanized antibody, single heavy chain antibody, single light chain antibody, scFv, and antigen-binding fragments thereof. 
     
     
         53 . The modified M1 macrophage or monocyte of  claim 49 , wherein the transmembrane domain of the CAR comprises a CD8 transmembrane domain. 
     
     
         54 . The modified M1 macrophage or monocyte of  claim 49 , wherein the intracellular domain of the CAR comprises CD80 and/or CD86 and/or CD40L and/or OX40L signaling domains. 
     
     
         55 . The modified M1 macrophage or monocyte of  claim 49 , wherein the intracellular domain of the CAR comprises a CD3 zeta intracellular domain. 
     
     
         56 . The modified M1 macrophage or monocyte of  claim 49 , wherein the modified cell exhibits targeted effector activity. 
     
     
         57 . The modified M1 macrophage or monocyte of  claim 56 , wherein the targeted effector activity is directed against a target cell comprising an antigen that specifically binds the antigen binding domain of the CAR. 
     
     
         58 . The modified M1 macrophage or monocyte of  claim 56 , wherein the targeted effector activity is selected from the group consisting of phagocytosis, cytotoxicity, and antigen presentation. 
     
     
         59 . The modified M1 macrophage or monocyte of  claim 49 , wherein the targeted effector activity is enhanced by inhibition of CD47 or signal-regulatory protein a activity. 
     
     
         60 . The modified M1 macrophage or monocyte of  claim 49 , further comprising an agent, wherein the agent is selected from the group consisting of a nucleic acid, a peptide and any combination thereof. 
     
     
         61 . The modified M1 macrophage or monocyte of  claim 49 , wherein the modified cell is genetically modified to express the CAR. 
     
     
         62 . A pharmaceutical composition comprising the modified M1 macrophage or monocyte of  claim 49  and a pharmaceutically acceptable carrier. 
     
     
         63 . The pharmaceutical composition of  claim 62 , wherein >75% of the modified cells in the pharmaceutical composition express the CAR or wherein the modified human monocytes in the population of cells are genetically modified to stably express the CAR. 
     
     
         64 . A method for stimulating an immune response to a target tumor cell or tumor tissue in a subject, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising the modified M1 macrophage or monocyte of  claim 49 .