Compositions and methods of NKG2D chimeric antigen receptor T cells for controlling triple-negative breast cancer
The invention includes a chimeric antigen receptor (CAR) specific for a natural-killer group 2, member D ligand (NKG2DL), compositions comprising the CAR, polynucleotides encoding the CAR, vectors comprising a polynucleotide encoding the CAR, and recombinant T cells comprising the CAR. The invention also includes methods of making a genetically modified T cell expressing a NKG2D-CAR wherein the expressed CAR comprises a NKG2D extracellular domain. The present invention also relates generally to the use of T cells engineered to express a CAR to treat cancer associated with NKG2D.
1 . A nucleic acid molecule encoding a chimeric antigen receptor (CAR), wherein the nucleic acid molecule comprises a nucleic acid sequence encoding a natural-killer group 2, member D (NKG2D) extracellular domain, a nucleic acid sequence encoding a CD27 costimulatory domain, and a nucleic acid sequence encoding a CD3 zeta signaling domain,
wherein the CAR comprises, from amino to carboxy terminus, the NKG2D extracellular domain, the CD27 costimulatory domain, and the CD3 zeta signaling domain, and
wherein the CAR comprises the amino acid sequence of SEQ ID NO: 15.
2 . The nucleic acid molecule of claim 1 , wherein the nucleic acid molecule comprises the nucleic acid sequence of SEQ ID NO: 14.
3 . A vector comprising the nucleic acid molecule of claim 1 .
4 . A chimeric antigen receptor (CAR) comprising a natural-killer group 2, member D (NKG2D) extracellular domain, a CD27 costimulatory domain, and a CD3 zeta signaling domain,
wherein the CAR comprises, from amino to carboxy terminus, the NKG2D extracellular domain, the CD27 costimulatory domain, and the CD3 zeta signaling domain, and
wherein the CAR comprises the amino acid sequence of SEQ ID NO: 15.
5 . The CAR of claim 4 , wherein the CAR is encoded by the nucleic acid sequence of SEQ ID NO: 14.
6 . A genetically modified T cell comprising the CAR of claim 4 .
7 . A method for treating cancer in a subject, the method comprising: administering to the subject an effective amount of the genetically modified T cell of claim 6 , thereby treating cancer in the subject.
8 . A genetically modified T cell comprising a nucleic acid molecule encoding a chimeric antigen receptor (CAR), wherein the nucleic acid molecule comprises a nucleic acid sequence encoding, from amino to carboxy terminus, a natural-killer group 2, member D (NKG2D) extracellular domain, a nucleic acid sequence encoding a CD27 costimulatory domain, and a nucleic acid sequence encoding a CD3 zeta signaling domain,
wherein the CAR comprises the amino acid sequence of SEQ ID NO: 15 or is encoded by the nucleic acid sequence of SEQ ID NO: 14.
9 . A genetically modified T cell comprising a nucleic acid molecule encoding a chimeric antigen receptor (CAR), wherein the nucleic acid molecule comprises the nucleic acid sequence of SEQ ID NO: 16 or encodes the amino acid sequence of SEQ ID NO: 17.
10 . A genetically modified T cell comprising a chimeric antigen receptor (CAR), wherein the CAR comprises the amino acid sequence of SEQ ID NO: 19 or is encoded by the nucleic acid sequence of SEQ ID NO: 18.
11 . A vector comprising a nucleic acid molecule encoding a chimeric antigen receptor (CAR), wherein the nucleic acid molecule comprises a nucleic acid sequence encoding, from amino to carboxy terminus, a natural-killer group 2, member D (NKG2D) extracellular domain, a nucleic acid sequence encoding a CD27 costimulatory domain, and a nucleic acid sequence encoding a CD3 zeta signaling domain,
wherein the nucleic acid molecule comprises the nucleic acid sequence of SEQ ID NO: 1.
12 . A method for treating triple negative breast cancer (TNBC) in a subject, the method comprising: administering to the subject an effective amount of a genetically modified T cell comprising a nucleic acid molecule encoding, from amino to carboxy terminus, a chimeric antigen receptor (CAR), wherein the nucleic acid molecule comprises a nucleic acid sequence of a natural-killer group 2, member D (NKG2D) extracellular domain, a nucleic acid sequence encoding a CD27 costimulatory domain, and a nucleic acid sequence encoding a CD3 zeta signaling domain, thereby treating TNBC in the subject.
13 . The method of claim 12 , wherein the subject is a human.
14 . The method of claim 12 , further comprising wherein the subject is administered a secondary treatment.
15 . The method of claim 14 , wherein the secondary treatment is selected from the group consisting of immune checkpoint blockade, chemotherapy, radiation, and surgery.