Compositions and methods for treating cancer with anti-CD19/CD22 immunotherapy
Chimeric antigen receptors containing CD19/CD22 or CD22/CD19 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. A method of generating a population of RNA-engineered cells comprising introducing an in vitro transcribed RNA or synthetic RNA into a cell, wherein the RNA comprises an isolated nucleic acid molecule encoding a chimeric antigen receptor (CAR) comprising at least one extracellular antigen binding domain comprising a CD19/CD22 antigen binding domain comprising the amino acid sequence of SEQ ID NO: 85 or 87, at least one transmembrane domain, and at least one intracellular signaling domain.
2. The method of claim 1 , wherein the at least one transmembrane domain comprises a transmembrane domain selected from the group consisting of: an alpha chain of a T-cell receptor, a beta chain of T-cell receptor, a zeta chain of a T-cell receptor, a CD8, a CD28, a CD3 epsilon, a CD45, a CD4, a CD5, a CD8, a CD9, a CD16, a CD22, a CD33, a CD37, a CD64, a CD80, a CD86, a CD134, a CD137 and a CD154.
3. The method of claim 1 , wherein the at least one extracellular antigen binding domain comprising the CD19/CD22 antigen binding domain, the at least one intracellular signaling domain, or both are connected to the at least one transmembrane domain by a linker or a spacer domain.
4. The method of claim 3 , wherein the linker or the spacer domain is derived from the extracellular domain of CD8 or CD28, and is linked to the at least one transmembrane domain.
5. The method of claim 1 , wherein the at least one extracellular antigen binding domain comprising the CD19/CD22 antigen binding domain is preceded by a leader nucleotide sequence encoding a leader peptide.
6. The method of claim 1 , wherein the at least one intracellular signaling domain comprises a costimulatory domain comprising a functional signaling domain of a protein selected from the group consisting of: OX40, CD70, CD27, CD28, CD5, ICAM-1, LFA-1, ICOS, DAP10, DAP12, 4-1BB, and a combination thereof.
7. The method of claim 1 , wherein the nucleic acid sequence encoding the CD19/CD22 tandem CAR is encoded by a nucleotide sequence comprising SEQ ID NO: 84 or 86, or a sequence with 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity thereof.
8. The method of claim 1 , wherein the at least one intracellular signaling domain comprises a costimulatory domain, a primary signaling domain, or any combination thereof.
9. The method of claim 1 , wherein the at least one intracellular signaling domain comprises a CD3 zeta intracellular domain.
10. A method of providing an anti-tumor immunity in a mammal comprising administering to the mammal an effective amount of an isolated cell comprising a vector comprising a nucleic acid molecule encoding a CD19/CD22 tandem chimeric antigen receptor (CAR) comprising at least one extracellular antigen binding domain comprising a CD19/CD22 antigen binding domain comprising the amino acid sequence of SEQ ID NO: 85 or 87, at least one transmembrane domain, and at least one intracellular signaling domain.
11. The method of claim 10 , wherein the at least one transmembrane domain comprises a transmembrane domain selected from the group consisting of: an alpha chain of a T-cell receptor, a beta chain of a T-cell receptor, a zeta chain of a T-cell receptor, a CD8, a CD28, a CD3 epsilon, a CD45, a CD4, a CD5, a CD8, a CD9, a CD16, a CD22, a CD33, a CD37, a CD64, a CD80, a CD86, a CD134, a CD137, and a CD154.
12. The method of claim 10 , wherein the at least one extracellular antigen binding domain comprising the CD19/CD22 antigen binding domain, the at least one intracellular signaling domain, or both are connected to the at least one transmembrane domain by a linker or a spacer domain.
13. The method of claim 12 , wherein the linker or the spacer domain is derived from the extracellular domain of CD8 or CD28, and is linked to the at least one transmembrane domain.
14. The method of claim 10 , wherein the at least one extracellular antigen binding domain comprising the CD19/CD22 antigen binding domain is preceded by a leader nucleotide sequence encoding a leader peptide.
15. The method of claim 10 , wherein the at least one intracellular signaling domain comprises a costimulatory domain comprising a functional signaling domain of a protein selected from the group consisting of: OX40, CD70, CD27, CD28, CD5, ICAM-1, LFA-1, ICOS, DAP10, DAP12, 4-1BB, and a combination thereof.
16. The method of claim 10 , wherein the CD19/CD22 tandem CAR is encoded by a nucleotide sequence comprising SEQ ID NO: 84 or 86, or a sequence with 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity thereof.
17. The method of claim 10 , wherein the at least one intracellular signaling domain comprises a costimulatory domain, a primary signaling domain, or any combination thereof.
18. The method of claim 10 , wherein the at least one intracellular signaling domain comprises a CD3 zeta intracellular domain.
19. A method of treating cancer in a mammal, comprising administering to the mammal a CD19/CD22 tandem CAR comprising at least one extracellular antigen binding domain comprising a CD19/CD22 antigen binding domain comprising the amino acid sequence of SEQ ID NO: 85 or 87, at least one transmembrane domain, and at least one intracellular signaling domain, in an amount effective to treat or prevent cancer in the mammal.
20. The method of claim 19 , wherein the at least one transmembrane domain comprises a transmembrane domain selected from the group consisting of: an alpha chain of a T-cell receptor, a beta chain of a T-cell receptor, a zeta chain of a T-cell receptor, a CD8, a CD28, a CD3 epsilon, a CD45, a CD4, a CD5, a CD8, a CD9, a CD16, a CD22, a CD33, a CD37, a CD64, a CD80, a CD86, a CD134, a CD137 and a CD154.
21. The method of claim 19 , wherein the at least one extracellular antigen binding domain comprising the CD19/CD22 antigen binding domain, the at least one intracellular signaling domain, or both are connected to the at least one transmembrane domain by a linker or a spacer domain.
22. The method of claim 21 , wherein the linker or the spacer domain is derived from the extracellular domain of CD8 or CD28, and is linked to the at least one transmembrane domain.
23. The method of claim 19 , wherein the at least one extracellular antigen binding domain comprising the CD19/CD22 antigen binding domain is preceded by a leader nucleotide sequence encoding a leader peptide.
24. The method of claim 19 , wherein the at least one intracellular signaling domain comprises a costimulatory domain comprising a functional signaling domain of a protein selected from the group consisting of: OX40, CD70, CD27, CD28, CD5, ICAM-1, LFA-1, ICOS, DAP10, DAP12, 4-1BB, and a combination thereof.
25. The method of claim 19 , wherein the CD19/CD22 tandem CAR is encoded by a nucleotide sequence comprising SEQ ID NO: 84 or 86, or a sequence with 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity thereof.
26. The method of claim 19 , wherein the at least one intracellular signaling domain comprises a costimulatory domain, a primary signaling domain, or any combination thereof.
27. The method of claim 19 , wherein the at least one intracellular signaling domain comprises a CD3 zeta intracellular domain.