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 - 26 . (canceled) 
     
     
         27 . A method of treating cancer comprising administering a pharmaceutical composition comprising
 (a) a population of CD14+ cells comprising a recombinant polynucleic acid, wherein the recombinant polynucleic acid comprises a sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises (i) an antigen binding domain;   (ii) a transmembrane domain; and (iii) an intracellular domain containing an intracellular signaling domain; and   (b) a pharmaceutically acceptable excipient   
       wherein the pharmaceutical composition stimulates tumor killing by T cells in a subject with cancer when the pharmaceutical composition is administered to the subject. 
     
     
         28 . The pharmaceutical composition of  claim 27 , wherein the CAR further comprises CD8a a hinge domain 
     
     
         29 . The pharmaceutical composition of  claim 27 , wherein the transmembrane domain comprises a CD8a transmembrane domain or a TLR4 transmembrane domain. 
     
     
         30 . The pharmaceutical composition of  claim 27 , wherein the intracellular domain comprises two or more signaling domains. 
     
     
         31 . The pharmaceutical composition of  claim 27 , wherein the intracellular domain comprises a CD3 zeta intracellular domain, an FcεRI intracellular domain, an FcγRI intracellular domain, a TLR4 intracellular domain, a TRIF intracellular domain, a TRADD intracellular domain, a MYD99 intracellular domain or a MYD88 intracellular domain. 
     
     
         32 . The pharmaceutical composition of  claim 31 , wherein the intracellular domain comprises an FcγRI intracellular domain or a TRIF intracellular domain. 
     
     
         33 . The pharmaceutical composition of  claim 31 , wherein the intracellular domain comprises a TRIF intracellular domain. 
     
     
         34 . The modified cell of  claim 31 , wherein the intracellular domain comprises a CD3 zeta intracellular domain. 
     
     
         35 . The pharmaceutical composition of  claim 27 , wherein the antigen binding domain is a single domain antibody (sdAb) or a single chain variable fragment (scFv). 
     
     
         36 . The pharmaceutical composition of  claim 27 , wherein the population of CD14+ cells is a population human CD14+ cells. 
     
     
         37 . The pharmaceutical composition of  claim 27 , wherein the population of CD14+ cells comprises CD16+ cells. 
     
     
         38 . The pharmaceutical composition of  claim 27 , wherein the population of CD14+ cells is a population of CD14+ monocytes, a population of CD14+ macrophages or a population of CD14+ dendritic cells. 
     
     
         39 . The pharmaceutical composition of  claim 38 , wherein the population of CD14+ cells is a population of CD14+ monocytes. 
     
     
         40 . The pharmaceutical composition of  claim 27 , wherein the population of CD14+ cells is from a leukapheresis. 
     
     
         41 . The pharmaceutical composition of  claim 27 , wherein the population of CD14+ cells is phagocytic. 
     
     
         42 . The pharmaceutical composition of  claim 27 , wherein the recombinant polynucleic acid is a viral vector. 
     
     
         43 . The pharmaceutical composition of  claim 27 , wherein the recombinant polynucleic acid is mRNA. 
     
     
         44 . The pharmaceutical composition of  claim 27 , wherein the antigen binding domain is an anti-HER2/neu binding domain. 
     
     
         45 . The pharmaceutical composition of  claim 27 , wherein the intracellular domain of the CAR is capable of inducing monocytic differentiation to M1 macrophages when the pharmaceutical composition is administered to a subject. 
     
     
         46 . The pharmaceutical composition of  claim 27 , wherein the population of CD14+ cells directly kill tumor cells and wherein the intracellular domain of the CAR is capable of inducing monocytic differentiation to dendritic cells when the pharmaceutical composition is administered to a subject with a tumor. 
     
     
         47 . The pharmaceutical composition of  claim 27 , wherein the pharmaceutical composition enhances or improves effector function of a T cell in a subject when the pharmaceutical composition is administered to the subject. 
     
     
         48 . The pharmaceutical composition of  claim 27 , wherein the pharmaceutical composition inhibits macrophage or macrophage related cells from promoting tumor growth when the pharmaceutical composition is administered to a subject with cancer. 
     
     
         49 . A method of treating cancer in a subject in need thereof, comprising administering the pharmaceutical composition of  claim 27  to the subject. 
     
     
         50 . The method of  claim 49 , wherein the cancer is a lymphoma. 
     
     
         51 . The method of  claim 49 , wherein the cancer is a solid tumor. 
     
     
         52 . The method of  claim 49 , wherein the cancer is a breast cancer. 
     
     
         53 . The method of  claim 49 , wherein the cancer is a metastatic cancer. 
     
     
         54 . The method of  claim 49 , wherein the cancer is a an ErbB-2-expressing cancer. 
     
     
         55 . The method of  claim 49 , wherein the method further comprises administering GM-CSF, IL-2 or an agent that blocks CD47 activity to the subject. 
     
     
         56 . A method of making the pharmaceutical composition of  claim 27 , the method comprising:
 (a) extracting a blood sample from a cancer patient;   (b) purifying peripheral blood mononuclear cells (PBMCs) from the blood sample;   (c) isolating CD14+ cells from the PBMCs, thereby obtaining isolated monocytes;   (d) introducing a recombinant polynucleic acid encoding a chimeric antigen receptor (CAR) into the isolated monocytes, thereby obtaining chimeric antigen receptor encoded monocytes, wherein the CAR comprises (i) an antigen binding domain; (ii) a transmembrane domain; and (iii) an intracellular domain containing an intracellular signaling domain;   (e) culturing the isolated monocytes in the presence of a cytokine; and   (f) preparing a pharmaceutical composition comprising the chimeric antigen receptor encoded monocytes from (e), wherein the pharmaceutical composition is suitable for infusing into the cancer patient.