Chimeric antigen receptors with an optimized hinge region
The present invention relates to multi-functional proteins which comprise (i) a signal peptide, (ii) a target specific recognition domain, (iii) a linker region, connecting domain (ii) and domain (iv) which comprises a specific modified hinge region of the human CD8 alpha-chain, and (iv) an effector domain. The present invention furthermore relates to nucleic acids encoding the proteins, expression constructs for expressing the protein in a host cell and host cells. The proteins of the invention are chimeric antigen receptors with an optimized linker or hinge region that are suitable for generating target-specific effector cells, for use as a medicament, in particular in the treatment of cancer and in adoptive, target-cell specific immunotherapy.
1. A nucleic acid sequence comprising nucleic acids encoding:
a linker region that does not contain a cysteine residue and is selected from the following:
a) the amino acid sequence of SEQ ID NO. 2,
b) an amino acid sequence having at least 95% sequence identity to SEQ ID NO. 2 under the proviso that amino acid residue 48 is a serine, and
c) an amino acid sequence that differs in one, two or three amino acid residues from the amino acid sequence of SEQ ID NO. 2 under the proviso that amino acid residue 48 is a serine.
2. The nucleic acid sequence of claim 1 , comprising: i) a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 2 or a sequence having 95% sequence identity to SEQ ID NO: 2, or ii) a nucleic acid sequence of SEQ ID NO: 8, a sequence complementary to SEQ ID NO: 8, or a sequence having at least 95% sequence identity to SEQ ID NO: 8.
3. The nucleic acid sequence of claim 1 , further comprising: i) a nucleic acid sequence encoding: a) the amino acid sequence of SEQ ID NO: 3 or 5 or a sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 3 or 5 or b) the amino acid residues 29-163 of SEQ ID NO: 3; or ii) the nucleic acid sequence of SEQ ID NO: 9 or 10, a sequence having 95% sequence identity to SEQ ID NO: 9 or 10, a sequence complementary to SEQ ID NO: 9 or 10, or a sequence having at least 95% sequence identity to the sequence complementary to SEQ ID NO: 9 or 10.
4. The nucleic acid sequence of claim 1 , further comprising: i) a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 6 or 7 or a sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 6 or 7, or ii) the nucleic acid sequence of SEQ ID NO: 11 or 12, a sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 11 or 12, a sequence complementary to the sequence of SEQ ID NO: 11 or 12, or a sequence having at least 95% sequence identity to the sequence complementary to SEQ ID NO: 11 or 12.
5. An expression construct comprising a nucleic acid sequence of claim 1 , and promoter and terminator sequences.
6. A host cell expressing a nucleic acid sequence of claim 1 , wherein the host cell is an effector cell of the immune system.
7. The host cell of claim 6 , wherein the effector cell of the immune system is a natural killer (NK) or a natural killer T (NKT) cell.
8. The nucleic acid sequence of claim 1 , further comprising: a nucleic acid sequence encoding a target specific recognition domain and a nucleic acid sequence encoding an effector domain, wherein:
A) the nucleic acid sequence encoding the target specific recognition domain comprises:
i) a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 6 or 7 or a sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 6 or 7, or
ii) the nucleic acid sequence of SEQ ID NO: 11 or 12, a sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 11 or 12, a sequence complementary to the sequence of SEQ ID NO: 11 or 12, or a sequence having at least 95% sequence identity to the sequence complementary to SEQ ID NO: 11 or 12; and
B) the nucleic acid sequence encoding the effector domain comprises:
i) a nucleic acid sequence encoding: a) the amino acid sequence of SEQ ID NO: 3 or 5 or a sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 3 or 5 or b) the amino acid residues 29-163 of SEQ ID NO: 3; or
ii) the nucleic acid sequence of SEQ ID NO: 9 or 10, a sequence having 95% sequence identity to SEQ ID NO: 9 or 10, a sequence complementary to SEQ ID NO: 9 or 10, or a sequence having at least 95% sequence identity to the sequence complementary to SEQ ID NO: 9 or 10.