MODIFIED NK-92 CELLS FOR TREATING CANCER
Provided herein are NK-92 cells expressing at least one CAR and at least one Fc receptor. Also provided are methods of treatment of a patient having or suspected of having a disease that is treatable with NK-92 cells, such as cancer, comprising administering to the patient NK-92-Fc-CAR.
1 . A method of treating a cancer in a patient in need thereof, the method comprising:
administering to the patient an effective amount of an NK-92 cell line comprising modified NK-92 cells;
wherein the modified NK-92 cells are each modified to express and display on the cell surface of the modified NK-92 cells at least one Fc receptor and at least one chimeric antigen receptor (CAR), wherein the CAR is capable of binding CD33 or CSPG-4 expressed on a surface of a cancer cell;
wherein the CAR has a scFv segment that is coupled via a hinge portion to a transmembrane domain and an intracellular signaling domain; and
wherein the scFv segment has the amino acid sequence of amino acids 21-266 of SEQ ID NO:11 or amino acids 21-281 SEQ ID NO:13.
2 . The method of claim 1 , wherein the CAR further comprises a signal peptide coupled to the N-terminus of the scFv segment, and wherein the signal peptide has the amino acid sequence of amino acids 1-20 of SEQ ID NO:11 or SEQ ID NO:13.
3 . The method of claim 1 , wherein the cancer is acute myeloid leukemia, breast cancer, a sarcoma, a neuroblastoma, or a mesothelioma.
4 . The method of claim 1 , wherein the hinge portion has the amino acid sequence of amino acids 267-327 of SEQ ID NO:11 or amino acids 284-347 SEQ ID NO:13.
5 . The method of claim 1 , wherein the transmembrane domain is a CD3zeta transmembrane domain.
6 . The method of claim 1 , wherein the intracellular signaling domain is a CD3zeta intracellular signaling domain.
7 . The method of claim 1 , wherein the CAR has the amino acid sequence of SEQ ID NO:11.
8 . The method of claim 1 , wherein the CAR is encoded by the nucleic acid sequence of SEQ ID NO:10.
9 . The method of claim 1 , wherein the CAR has the amino acid sequence of SEQ ID NO:13.
10 . The method of claim 1 , wherein the CAR is encoded by the nucleic acid sequence of SEQ ID NO:12.
11 . The method of claim 1 , wherein the Fc receptor is FcγRIII-A (CD16) or a CD16 polypeptide having a valine at position 158 of the mature form of the CD16.
12 . The method of claim 1 , wherein the Fc receptor has the amino acid sequence of SEQ ID NO:2.
13 . The method of claim 1 , wherein the modified NK-92 cells are modified to express a cytokine.
14 . The method of claim 13 , wherein the cytokine is IL-2 or IL15, optionally modified to target the endoplasmic reticulum.
15 . The method of claim 1 , wherein the effective amount is at least about 1×10 8 cells.
16 . A modified NK92-cell, wherein
(1) the modified NK-92 cells are modified to each express at least one Fc receptor and at least one chimeric antigen receptor (CAR), such that the at least one Fc receptor and the at least one CAR are displayed on the cell surface of the modified NK-92 cells;
(2) the CAR has a scFv segment that is coupled via a hinge portion to a transmembrane domain and an intracellular signaling domain, and wherein the CAR is capable of binding CD33 or CSPG-4 expressed on a surface of a cancer cell; and
(3) the scFv segment has the amino acid sequence of amino acids 21-266 of SEQ ID NO:11 or amino acids 21-281 SEQ ID NO:13.
17 . The modified NK92-cell of claim 16 , wherein the CAR is encoded by the nucleic acid sequence of SEQ ID NO:10 or wherein the CAR is encoded by the nucleic acid sequence of SEQ ID NO:12.
18 . The modified NK92-cell of claim 16 , wherein the transmembrane domain is a CD3zeta transmembrane domain, and/or wherein the intracellular signaling domain is a CD3zeta intracellular signaling domain.
19 . The modified NK92-cell of claim 16 , wherein the modified NK-92 cells are modified to express a cytokine.
20 . The modified NK92-cell of claim 16 , wherein the Fc receptor is FcγRIII-A (CD16) or a CD16 polypeptide having a valine at position 158 of the mature form of the CD16.