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 population of modified human monocytes 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, wherein >75% of the modified human monocytes in the population of cells express the CAR or wherein the modified human monocytes in the population of cells are genetically modified to stably express the CAR, and wherein the modified human monocytes replicate in vivo resulting in long-term persistence that can lead to sustained tumor control. 
     
     
         22 . The population of  claim 21 , wherein the antigen binding domain comprises an anti-HER2 antigen binding domain. 
     
     
         23 . The population 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 
     
     
         24 . The population of  claim 21 , wherein the transmembrane domain of the CAR comprises a CD8 transmembrane domain. 
     
     
         25 . The population of  claim 21 , wherein the intracellular domain of the CAR comprises CD80 and/or CD86 and/or CD40L and/or OX40L signaling domains. 
     
     
         26 . The population of  claim 21 , wherein the intracellular domain of the CAR comprises a CD3 zeta intracellular domain. 
     
     
         27 . The population of  claim 21 , wherein the modified human monocytes exhibit targeted effector activity. 
     
     
         28 . The population of  claim 27 , wherein the targeted effector activity is directed against a target cell comprising an antigen that specifically binds the antigen binding domain of the CAR. 
     
     
         29 . The population of  claim 27 , wherein the targeted effector activity is selected from the group consisting of phagocytosis, cytotoxicity, and antigen presentation. 
     
     
         30 . The population of  claim 27 , wherein the targeted effector activity is enhanced by inhibition of CD47 or signal-regulatory protein a activity. 
     
     
         31 . The population 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. 
     
     
         32 . A pharmaceutical composition comprising the population of modified human monocytes of  claim 51  and a pharmaceutically acceptable carrier. 
     
     
         33 . 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 population of modified human monocytes of  claim 51 . 
     
     
         34 . A method of modifying a population of human monocytes, comprising: introducing a chimeric antigen receptor (CAR) into the population of human monocytes, wherein the CAR comprises an antigen binding domain, a transmembrane domain and an intracellular domain of a stimulatory and/or co-stimulatory molecule, wherein >75% of the monocytes in the population of modified human monocytes express the CAR or wherein the modified human monocytes in the population of cells are genetically modified to stably express the CAR, and wherein the modified human monocytes replicate in vivo resulting in long-term persistence that can lead to sustained tumor control. 
     
     
         35 . The method of  claim 34 , wherein introducing the CAR into the population of human monocytes comprises introducing a nucleic acid sequence encoding the CAR. 
     
     
         36 . The method of  claim 34 , wherein introducing the nucleic acid sequence comprises introducing an mRNA encoding the CAR. 
     
     
         37 . The method of  claim 34 , wherein introducing the nucleic acid sequence comprises transducing the cell with a viral vector comprising a nucleic acid sequence encoding the CAR. 
     
     
         38 . The method of  claim 34 , wherein the modified human monocytes exhibit targeted effector activity. 
     
     
         39 . The method of  claim 38 , wherein the targeted effector activity is selected from the group consisting of phagocytosis, cytotoxicity, and antigen presentation. 
     
     
         40 . The method of  claim 34 , further comprising inhibiting CD47 or signal-regulatory protein a activity to enhance the targeted effector activity. 
     
     
         41 . The method of  claim 40 , wherein inhibiting CD47 activity comprises contacting the human monocyte with a blocking anti-CD47 antibody. 
     
     
         42 . The method of  claim 34 , wherein the antigen binding domain comprises an anti-HER2 antigen binding domain. 
     
     
         43 . The method of  claim 42 , 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. 
     
     
         44 . A pharmaceutical composition comprising:
 (a) a population of modified human macrophages and/or monocytes comprising a chimeric antigen receptor (CAR), wherein >75% of the macrophages and/or monocytes in the population of modified human macrophages and/or monocytes express the CAR or wherein the modified human macrophages and/or monocytes in the population of modified human macrophages and/or monocytes are genetically modified to stably express the CAR, and   (b) a pharmaceutically acceptable excipient.   
     
     
         45 . The pharmaceutical composition of  claim 44 , wherein the CAR comprises an antigen binding domain, a transmembrane domain and an intracellular domain of a stimulatory and/or co-stimulatory molecule. 
     
     
         46 . The pharmaceutical composition of  claim 44 , wherein the antigen binding domain comprises an anti-HER2 antigen binding domain. 
     
     
         47 . The pharmaceutical composition of  claim 45 , 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. 
     
     
         48 . The pharmaceutical composition of  claim 45 , wherein the transmembrane domain of the CAR comprises a CD8 transmembrane domain. 
     
     
         49 . The pharmaceutical composition of  claim 45 , wherein the intracellular domain of the CAR comprises CD80 and/or CD86 and/or CD40L and/or OX40L signaling domains. 
     
     
         50 . The pharmaceutical composition of  claim 45 , wherein the intracellular domain of the CAR comprises a CD3 zeta intracellular domain. 
     
     
         51 . The pharmaceutical composition of  claim 44 , wherein the modified human macrophages and/or monocytes exhibit targeted effector activity. 
     
     
         52 . The pharmaceutical composition of  claim 51 , wherein the targeted effector activity is directed against a target cell comprising an antigen that specifically binds the antigen binding domain of the CAR. 
     
     
         53 . The pharmaceutical composition of  claim 51 , wherein the targeted effector activity is selected from the group consisting of phagocytosis, cytotoxicity, and antigen presentation. 
     
     
         54 . The pharmaceutical composition of  claim 51 , wherein the targeted effector activity is enhanced by inhibition of CD47 or signal-regulatory protein a activity. 
     
     
         55 . The pharmaceutical composition of  claim 44 , further comprising an agent, wherein the agent is selected from the group consisting of a nucleic acid, a peptide and any combination thereof. 
     
     
         56 . 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 44 . 
     
     
         57 . A method of modifying a population of human cells, comprising: introducing a chimeric antigen receptor (CAR) into the population of human macrophages and/or monocytes, wherein >75% of the macrophages and/or monocytes in the population of human macrophages and/or monocytes express the CAR or wherein the human macrophages and/or monocytes in the population of human macrophages and/or monocytes are genetically modified to stably express the CAR. 
     
     
         58 . The method of  claim 57 , wherein introducing the CAR into the population of human macrophages and/or monocytes comprises introducing a nucleic acid sequence encoding the CAR. 
     
     
         59 . The method of  claim 58 , wherein introducing the nucleic acid sequence comprises introducing an mRNA encoding the CAR. 
     
     
         60 . The method of  claim 58 , wherein introducing the nucleic acid sequence comprises transducing the population of human macrophages and/or monocytes with a viral vector comprising a nucleic acid sequence encoding the CAR. 
     
     
         61 . The method of  claim 57 , wherein the CAR comprises an antigen binding domain, a transmembrane domain and an intracellular domain of a stimulatory and/or co-stimulatory molecule. 
     
     
         62 . The method of  claim 61 , wherein the antigen binding domain comprises an anti-HER2 antigen binding domain. 
     
     
         63 . The method of  claim 62 , 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. 
     
     
         64 . The method of  claim 61 , wherein the intracellular domain of the CAR comprises CD80 and/or CD86 and/or CD40L and/or OX40L signaling domains. 
     
     
         65 . The method of  claim 57 , wherein the modified human macrophages and/or monocytes exhibit targeted effector activity. 
     
     
         66 . The method of  claim 57 , wherein the targeted effector activity is directed against a target cell comprising an antigen that specifically binds the antigen binding domain of the CAR. 
     
     
         67 . The method of  claim 65 , wherein the targeted effector activity is selected from the group consisting of phagocytosis, cytotoxicity, and antigen presentation. 
     
     
         68 . The method of  claim 65 , wherein the targeted effector activity is enhanced by inhibition of CD47 or signal-regulatory protein a activity.