Compositions and methods for treating cancer with anti-mesothelin immunotherapy
Chimeric antigen receptors containing mesothelin antigen binding domains are disclosed. Nucleic acids, recombinant expression vectors, host cells, antigen binding fragments, and pharmaceutical compositions, relating to the chimeric antigen receptors are also disclosed. Methods of treating or preventing cancer in a subject, and methods of making chimeric antigen receptor T cells are also disclosed.
1. An isolated nucleic acid molecule encoding a chimeric antigen receptor (CAR) comprising at least one extracellular antigen binding domain comprising a mesothelin antigen binding domain encoded by a nucleotide sequence comprising SEQ ID NO. 3, 5, or 7, at least one transmembrane domain, and at least one intracellular signaling domain.
2. A chimeric antigen receptor (CAR) encoded by the isolated nucleic acid molecule of claim 1 .
3. A vector comprising the isolated nucleic acid molecule of claim 1 .
4. The vector of claim 3 , wherein the vector is selected from the group consisting of a DNA vector, an RNA vector, a plasmid vector, a cosmid vector, a herpes virus vector, a measles virus vector, a lentivirus vector, adenoviral vector, a retrovirus vector, and a combination thereof.
5. An isolated cell comprising the vector of claim 3 .
6. A method of making a cell comprising transducing an isolated T cell with the vector of claim 3 .
7. A pharmaceutical composition comprising an anti-tumor effective amount of a population of human T cells, wherein the T cells comprise a nucleic acid sequence that encodes a chimeric antigen receptor (CAR), wherein the CAR comprises at least one extracellular antigen binding domain comprising a mesothelin antigen binding domain comprising the amino acid sequence of SEQ ID NO. 4, 6, or 8, at least one linker domain, at least one transmembrane domain, and at least one intracellular signaling domain, wherein the T cells are T cells of a human subject having a cancer.
8. A method of treating cancer in a subject in need thereof, the method comprising administering to the subject a pharmaceutical composition comprising an anti-tumor effective amount of a population of T cells, wherein the T cells comprise a nucleic acid sequence that encodes a chimeric antigen receptor (CAR), wherein the CAR comprises at least one extracellular antigen binding domain comprising a mesothelin antigen binding domain comprising the amino acid sequence of SEQ ID NO. 4, 6, or 8, at least one linker or spacer domain, at least one transmembrane domain, and at least one intracellular signaling domain, wherein the T cells are T cells of the subject having cancer.
9. The method of claim 8 , wherein the at least one transmembrane domain comprises a transmembrane domain of a protein selected from the alpha, beta or zeta chain of the T-cell receptor, CD8, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, CD154, or any combination thereof.
10. The method of claim 8 , wherein the encoded at least one mesothelin antigen binding domain, the at least one intracellular signaling domain, or both are connected to the transmembrane domain by the at least one linker or spacer domain.
11. The method of claim 10 , wherein the encoded at least one linker or spacer domain is from the extracellular domain of CD8 or CD28, and is linked to a transmembrane domain.
12. The method of claim 8 , wherein the nucleic acid sequence encoding the extracellular mesothelin antigen binding domain comprises a nucleic sequence comprising SEQ ID NO, 3, 5, or 7, or a sequence with 85%, 90%, 95%, 96%, 97%, 98% or 99% identity thereof.
13. The method of claim 8 , wherein the encoded at least one intracellular signaling domain comprises a signaling domain of CD3 zeta.
14. The method of claim 8 , wherein the encoded at least one intracellular signaling domain comprises a costimulatory domain, a primary signaling domain, or any combination thereof.
15. The method of claim 14 , wherein the costimulatory domain comprises a functional signaling domain of OX40, CD70, CD27, CD28, CD5, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), DAP10, DAP12, or 4-1BB (CD137), or any combination thereof.
16. The method of claim 8 , wherein the cancer is a hematological cancer.
17. The method of claim 16 , wherein the hematological cancer is leukemia, lymphoma, or multiple myeloma.
18. The method of claim 17 , wherein the leukemia is chronic lymphocytic leukemia (CLL), acute lymphocytic leukemia (ALL), or chronic myelogenous leukemia (CIVIL).
19. The method of claim 17 , wherein the lymphoma is mantle cell lymphoma, non-Hodgkin's lymphoma, or Hodgkin's lymphoma.
20. The method of claim 8 , wherein the cancer is an oral and pharynx cancer, a digestive system cancer, a respiratory system cancer, a bone and joint cancer, a soft tissue cancer, a skin cancer, a pediatric cancer, a cancer of the central nervous system, a cancer of the breast, a cancer of the genital system, a cancer of the urinary system, a cancer of the eye and orbit, a cancer of the endocrine system, a cancer of the brain, or a combination thereof.
21. An isolated nucleic acid molecule encoding a chimeric antigen receptor (CAR) comprising at least one extracellular antigen binding domain comprising a mesothelin antigen binding domain, at least one transmembrane domain, and at least one intracellular signaling domain, wherein the mesothelin antigen binding domain comprises the amino acid sequence of SEQ ID NO. 4, 6, or 8.
22. The isolated nucleic acid molecule of claim 21 , wherein the mesothelin antigen binding domain comprise the amino acid sequence of SEQ ID NO. 4, wherein the at least one transmembrane domain comprises a CD8 transmembrane domain, and wherein the at least one intracellular signaling domain comprises a signaling domain of CD3 zeta and a signaling domain of 4-1BB (CD137).
23. The isolated nucleic acid molecule of claim 22 , wherein the CAR comprises the amino acid sequence of SEQ ID NO. 14.
24. The isolated nucleic acid molecule of claim 21 , wherein the mesothelin antigen binding domain comprise the amino acid sequence of SEQ ID NO. 6, wherein the at least one transmembrane domain comprises a CD8 transmembrane domain, and wherein the at least one intracellular signaling domain comprises a signaling domain of CD3 zeta and a signaling domain of 4-1BB (CD137).
25. The isolated nucleic acid molecule of claim 24 , wherein the CAR comprises the amino acid sequence of SEQ ID NO. 18.
26. The isolated nucleic acid molecule of claim 21 , wherein the mesothelin antigen binding domain comprise the amino acid sequence of SEQ ID NO. 8, wherein the at least one transmembrane domain comprises a CD8 transmembrane domain, and wherein the at least one intracellular signaling domain comprises a signaling domain of CD3 zeta and a signaling domain of 4-1BB (CD137).
27. The isolated nucleic acid molecule of claim 26 , wherein the CAR comprises the amino acid sequence of SEQ ID NO. 16.