T cell modification and use thereof
This invention relates to modified T cells that inducibly express a bioactive molecule, such as IL-7, and constitutively expresses an antigen receptor, such as a T cell receptor or chimeric antigen receptor that binds to a tumour antigen. The modified T cells may comprise a nucleic acid construct that comprises a first nucleotide sequence encoding the bioactive molecule, a second nucleotide sequence encoding the antigen receptor; an inducible promoter operably linked to the first nucleotide sequence and a constitutive promoter operably linked to the second nucleotide. Nucleic acid constructs and vectors are provided, as well as T cells comprising such constructs and vectors and therapeutic methods and uses thereof.
1. A nucleic acid construct comprising:
(i) a first nucleotide sequence encoding IL-7,
(ii) a second nucleotide sequence encoding an antigen receptor, wherein the antigen receptor is a chimeric antigen receptor (CAR) that binds to a tumor antigen, or is a T cell receptor (TCR) that binds to an MHC-displayed peptide fragment of the tumor antigen;
(iii) an inducible promoter operably linked to the first nucleotide sequence and
(iv) a constitutive promoter operably linked to the second nucleotide.
2. The nucleic acid construct according to claim 1 wherein the expression from the inducible promoter is induced by the activation of T cells.
3. The nucleic acid construct according to claim 1 wherein the inducible promoter comprises a nuclear factor of activated T cells (NFAT) transcriptional response element (TRE).
4. The nucleic acid construct according to claim 3 wherein the NFAT TRE comprises the nucleic acid sequence of SEQ ID NO: 14 or a variant thereof.
5. The nucleic acid construct according to claim 3 wherein the inducible promoter comprises three or more copies of the NFAT TRE.
6. The nucleic acid construct according to claim 1 wherein the nucleic acid construct comprises the sequence of SEQ ID NO: 1.
7. The nucleic acid construct according to claim 1 wherein the constitutive promoter is the Human elongation factor-1 alpha promoter.
8. The nucleic acid construct according to claim 1 wherein the IL-7 is human IL-7.
9. The nucleic acid construct according to claim 1 wherein the antigen receptor is a T cell receptor (TCR).
10. The nucleic acid construct according to claim 9 wherein the TCR is an affinity enhanced TCR.
11. The nucleic acid construct according to claim 9 wherein the TCR comprises the amino acid sequence of any one of SEQ ID NOs: 5, 6 or 11.
12. The nucleic acid construct according to claim 1 wherein the antigen receptor is a chimeric antigen receptor (CAR).
13. The nucleic acid construct according to claim 12 wherein the tumor antigen is NY-ESO1, PRAME, alpha-fetoprotein (AFP), MAGE A4, MAGE A1, MAGE A10 or MAGE B2.
14. The nucleic acid construct according to claim 1 wherein the first nucleotide sequence is configured for expression in a first direction and the second nucleotide sequence is configured for expression in a first direction.
15. A vector comprising a nucleic acid construct according to claim 1 .
16. A viral particle comprising a vector according to claim 15 .
17. A population of T cells each comprising a nucleic construct-according to claim 1 .
18. A pharmaceutical composition comprising a population of T cells according to claim 17 a pharmaceutically acceptable excipient.
19. The nucleic acid construct according to claim 10 wherein the tumor antigen is NY-ESO1, PRAME, alpha-fetoprotein (AFP), MAGE A4, MAGE A1, MAGE A10 or MAGE B2.