Cell-mediated transient delivery of immune-enhancing molecules into the tumor microenvironment
A recombinant natural killer (NK) cell or T-cell composition is transfected with a nucleic acid encoding i) a homing receptor; ii) an antigen binding protein (ABP) or a chimeric antigen receptor (CAR) that specifically binds a target antigen; iii) an Fc Receptor; and/or iv) a secreted immune modulator selected from a TGFβ inhibitor and/or IL-12, where the recombinant cell is gamma (γ)-irradiated conferring inhibition of cell proliferation with transient activity of the transfected molecules including the secreted immune modulators for up to 72 hours.
1. A recombinant natural killer (NK)-92 cell transfected with a plasmid expression vector and gamma (γ)-irradiated,
wherein the plasmid expression vector comprises a nucleic acid that encodes i) a homing receptor; ii) an antigen binding protein (ABP) or a chimeric antigen receptor (CAR) that specifically binds a target antigen; iii) an Fc Receptor; and iv) a secreted immune modulator, wherein the secreted immune modulator is a TGFβ inhibitor,
wherein the homing receptor is selected from the group consisting of a G protein-coupled receptor, a chemokine receptor, a cytokine receptor, a cell adhesion molecule, a selectin, and an integrin,
wherein the TGFβ inhibitor has the amino acid sequence of SEQ ID NO:64 or SEQ ID NO:66 and
wherein the nucleic acid is operably linked to a promoter.
2. The recombinant NK-92 cell of claim 1 , wherein the recombinant NK-92 cell is irradiated at between 2.5 to 20 Gy.
3. The recombinant NK-92 cell of claim 1 , wherein the modified NK-92 cell is irradiated at 5, 10, or 15 Gy.
4. The recombinant NK-92 cell of claim 1 , wherein the nucleic acid is a quadricistronic vector.
5. The recombinant NK-92 cell of claim 1 , wherein the antigen binding protein is selected from a protein or antibody that binds to CD19, CD20, GD2, HER-2, CD30, EGFR, FAP, CD33, CD123, PD-L1, IGF1R, CSPG4, or B7-H4.
6. The recombinant NK-92 cell of claim 1 , wherein the antigen binding protein binds to PD-L1.
7. The recombinant NK-92 cell of claim 1 , wherein the Fc Receptor is CD16 or a high affinity CD16.
8. The recombinant NK-92 cell of claim 1 , wherein the CAR specifically binds PD-L1.
9. The recombinant NK-92 cell of claim 1 , wherein the recombinant NK-92 cell has inhibited cell replication after exposure to the gamma radiation and wherein the secreted TGFβ inhibitor is transiently active up to about 72 hours.
10. A composition comprising the recombinant NK-92 cell of claim 1 and a pharmaceutically acceptable excipient.
11. A kit comprising the recombinant NK-92 cell of claim 1 and instructions for use.