Compositions and methods to modify cells for therapeutic objectives

Abstract

The present disclosure provides compositions and methods that rapidly and selectively modify cells of the immune system to achieve therapeutic objectives. The methods can be practiced in vivo and any cell type that expresses a known marker can be targeted for a therapeutic objective. The present disclosure provides compositions and methods that rapidly and selectively modify cells of the immune system to achieve therapeutic objectives. The methods can be practiced in vivo and any cell type that expresses or is associated with a known marker can be targeted for a therapeutic objective by the modified cell.

1 - 42 . (canceled) 
     
     
         43 . A synthetic nanocarrier comprising (i) a lipid-coated porous nanoparticle (ii) a lymphocyte-directing agent extending from the surface of the nanoparticle; and (iii) a polynucleotide encoding a chimeric antigen receptor (CAR) targeting agent within the pores of the nanoparticle. 
     
     
         44 . A synthetic nanocarrier of  claim 43  further comprising an endosomal release agent extending from the surface of the nanoparticle and (ii) a nuclear localization signal within the pores of the nanoparticle. 
     
     
         45 . A synthetic nanocarrier of  claim 43  wherein the synthetic nanocarrier comprises a liposome, polymeric particle, metallic particle, polymeric micelle, polyethyleneimine (PEI)/DNA complex, or a combination thereof. 
     
     
         46 . A synthetic nanocarrier of  claim 43  wherein the lymphocyte-directing agent selectively binds to lymphocytes in vivo. 
     
     
         47 . A synthetic nanocarrier of  claim 43  wherein the lymphocyte-directing agent selectively binds T cells, NK cells, monocytes, macrophages, B cells, hematopoietic stem cells or a combination thereof. 
     
     
         48 . A synthetic nanocarrier of  claim 43  wherein the lymphocyte-directing agent selectively binds CCR7; CD3; CD4; CD5; CD8; CD16; CD19; CD20; CD21; CD22;
 CD25; CD28; CD35; CD40; CD45RA; CD45RO; CD52; CD62L; CD80; CD95; CD127; 
 or CD137. 
 
     
     
         49 . A synthetic nanocarrier of  claim 43  wherein the lymphocyte-directing agent selectively binds CD8. 
     
     
         50 . A synthetic nanocarrier of  claim 43  wherein the lymphocyte-directing agent comprises a binding domain selected from a lymphocyte receptor ligand, lymphocyte receptor antibody, lymphocyte receptor peptide aptamer, lymphocyte receptor nucleic acid aptamer, lymphocyte receptor spiegelmer or a combination thereof. 
     
     
         51 . A synthetic nanocarrier of  claim 49  wherein the binding domain consists of an ScFv fragment of a T-cell a chain antibody; T-cell β chain antibody; T-cell γ chain antibody; T-cell Δ chain antibody; CCR7 antibody; CD3 antibody; CD4 antibody; CD5 antibody; CD7 antibody; CD8 antibody; CD11b antibody; CD11c antibody; CD16 antibody; CD19 antibody; CD20 antibody; CD21 antibody; CD22 antibody; CD25 antibody; CD28 antibody; CD34 antibody; CD35 antibody; CD40 antibody; CD45RA antibody; CD45RO antibody; CD52 antibody; CD56 antibody; CD62L antibody; CD68 antibody; CD80 antibody; CD95 antibody; CD117 antibody; CD127 antibody; CD133 antibody; CD137 (4-1BB) antibody; CD163 antibody; F4/80 antibody; IL-4Rα antibody;
 Sca-1 antibody; CTLA-4 antibody; GITR antibody GARP antibody; LAP antibody; 
 granzyme B antibody; LFA-1 antibody; transferrin antibody; or perforin antibody. 
 
     
     
         52 . A synthetic nanocarrier of  claim 43  wherein the polynucleotide is a plasmid, a minicircle plasmid, or an mRNA molecule. 
     
     
         53 . A synthetic nanocarrier of  claim 52  wherein the polynucleotide is a minicircle plasmid encoding a hyperactive transposase. 
     
     
         54 . A synthetic nanocarrier of  claim 43  comprising a S/MAR element, a PiggyBac transposase-containing plasmid, a Sleeping Beauty transposase-containing plasmid; a homo sapiens transposon-derived Buster1 transposase-like protein gene; a human endogenous retrovirus H protease/integrase-derived ORF1; a homo sapiens Cas-Br-M (murine) ecotropic retroviral transforming sequence; a homo sapiens endogenous retroviral sequence K; a homo sapiens endogenous retroviral family W sequence; a homo sapiens LINE-1 type transposase domain; or a homo sapiens pogo transposable element. 
     
     
         55 . A synthetic nanocarrier of  claim 43  wherein the CAR targeting agent comprises a binding domain for a marker associated with an unwanted cell type. 
     
     
         56 . A synthetic nanocarrier of  claim 55  wherein the unwanted cell type is a cancer cell. 
     
     
         57 . A synthetic nanocarrier of  claim 43  wherein the CAR targeting agent is a surface antigen receptor or a receptor for an intracellular antigen presented by a Major Histocompatibility Complex antigen-presenting pathway. 
     
     
         58 . A synthetic nanocarrier of  claim 44  wherein the endosomal release agent is selected from any one of SEQ ID NOs.29-50. 
     
     
         59 . A synthetic nanocarrier of  claim 44  wherein the NLS is selected from any of SEQ ID NOs. 51-93. 
     
     
         60 . A method for treating a disease associated with an antigen, the method comprising: administering to a subject in need thereof, a synthetic nanocarrier comprising (i) a lipid-coated porous nanoparticle with an endosomal release agent extending from the surface of the nanoparticle (ii) a lymphocyte-directing agent extending from the surface of the nanoparticle; (iii) a polynucleotide encoding a chimeric antigen receptor (CAR) targeting agent within the pores of the nanoparticle; and (iv) a nuclear localization signal within the pores of the nanoparticle thereby treating the disease in the subject. 
     
     
         61 . A method of selectively transfecting lymphocytes in a subject in vivo, the method comprising:
 contacting lymphocytes within the subjection with a synthetic nanocarrier comprising (i) a lipid-coated porous nanoparticle with an endosomal release agent extending from the surface of the nanoparticle (ii) a lymphocyte-directing agent extending from the surface of the nanoparticle; (iii) a polynucleotide encoding a chimeric antigen receptor (CAR) targeting agent within the pores of the nanoparticle; and (iv) a nuclear localization signal within the pores of the nanoparticle thereby transfecting lymphocytes in the subject in vivo.