Chimeric GMCSF-IL18 receptor
The present invention provides chimeric cytokine receptors, particularly chimeric cytokine receptors that can be activated in tumor microenvironment, and their uses in tumor immunotherapy (e.g., adoptive cell therapy). The present invention further provides methods of genetically modifying therapeutic cells resulting in an enhanced immune response against a target antigen. The application further provides therapeutic cells that express said chimeric cytokine receptors and methods for treating patients using the modified therapeutic cells.
1 . A polynucleotide encoding a chimeric cytokine receptor, said chimeric cytokine receptor comprising two polypeptides, each comprising an extracellular domain of granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor, or a functional portion thereof, a transmembrane domain, and an intracellular domain of interleukin-18 receptor (IL-18) receptor, or a functional portion thereof,
wherein the chimeric cytokine receptor improves effector function when expressed in an immune cell expressing a chimeric antigen receptor (CAR), an antigen specific T cell receptor (TCR) and/or a bispecific antibody, relative to an immune cell not expressing the chimeric cytokine receptor.
2 . The polynucleotide of claim 1 , wherein the chimeric cytokine receptor comprises
(i) a first polypeptide comprising an extracellular region of GM-CSF receptor α chain, or a functional portion thereof, a first transmembrane region, and an intracellular region of IL-18 receptor α chain, or a functional portion thereof; and
(ii) a second polypeptide comprising an extracellular region of GM-CSF receptor β chain, or a functional portion thereof, a second transmembrane region, and an intracellular region of IL-18 receptor β chain, or a functional portion thereof.
3 . The polynucleotide of claim 2 , wherein the extracellular region of GM-CSF receptor α chain comprises the amino acid sequence of SEQ ID NO: 1, or an amino acid sequence having at least 80% sequence identity thereof; or the sequence encoding the extracellular region of GM-CSF receptor α chain comprises the nucleotide sequence of SEQ ID NO: 2, or a nucleotide sequence having at least 80% sequence identity thereof.
4 . The polynucleotide of claim 2 , wherein the intracellular region of IL-18 receptor α chain comprises the amino acid sequence of SEQ ID NO: 5, or an amino acid sequence having at least 80% sequence identity thereof, or the sequence encoding the intracellular region of IL-18 receptor α chain comprises the nucleotide sequence of SEQ ID NO: 6, or a nucleotide sequence having at least 80% sequence identity thereof.
5 . The polynucleotide of claim 2 , wherein the extracellular region of GM-CSF receptor β chain comprises the amino acid sequence of SEQ ID NO: 7, or an amino acid sequence having at least 80% sequence identity thereof; or the sequence encoding the extracellular region of GM-CSF receptor β chain comprises the nucleotide sequence of SEQ ID NO: 8, or a nucleotide sequence having at least 80% sequence identity thereof.
6 . The polynucleotide of claim 2 , wherein the intracellular region of IL-18 receptor β chain comprises the amino acid sequence of SEQ ID NO: 11, or an amino acid sequence having at least 80% sequence identity thereof; or the sequence encoding the intracellular region of IL-18 receptor β chain comprises the nucleotide sequence of SEQ ID NO: 12, or a nucleotide sequence having at least 80% sequence identity thereof.
7 . The polynucleotide of claim 2 , wherein at least one of the transmembrane regions is derived from a transmembrane domain of IL-18 receptor or GM-CSF receptor.
8 . The polynucleotide of claim 2 , wherein the first transmembrane region comprises a transmembrane region of IL-18 receptor α chain.
9 . The polynucleotide of claim 8 , wherein the transmembrane region of IL-18 receptor α chain comprises the amino acid sequence of SEQ ID NO: 3, or an amino acid sequence having at least 80% sequence identity thereof; or the sequence encoding the transmembrane region of IL-18 receptor α chain comprises the nucleotide sequence of SEQ ID NO: 4, or a nucleotide sequence having at least 80% sequence identity thereof.
10 . The polynucleotide of claim 2 , wherein the second transmembrane region comprises a transmembrane region of IL-18 receptor β chain.
11 . The polynucleotide of claim 10 , wherein the transmembrane region of IL-18 receptor β chain comprises the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence having at least 80% sequence identity thereof, or the sequence encoding the transmembrane region of IL-18 receptor β chain comprises the nucleotide sequence of SEQ ID NO: 10, or a nucleotide sequence having at least 80% sequence identity thereof.
12 . The polynucleotide of claim 2 , wherein the first polypeptide further comprises a first leader sequence and/or the second polypeptide further comprises a second leader sequence.
13 . The polynucleotide of claim 12 , wherein the first leader sequence is derived from a leader sequence of GM-CSF receptor α chain.
14 . The polynucleotide of claim 12 , wherein the first leader sequence comprises the amino acid sequence of SEQ ID NO: 13, SEQ ID NO: 44, or SEQ ID NO: 46, or an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 13, SEQ ID NO: 44, or SEQ ID NO: 46; or the sequence encoding the first leader sequence comprises the nucleotide sequence of SEQ ID NO: 14, SEQ ID NO: 45, or SEQ ID NO: 47, or a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 14, SEQ ID NO: 45, or SEQ ID NO: 47.
15 . The polynucleotide of claim 12 , wherein the second leader sequence is derived from a leader sequence of GM-CSF receptor β chain.
16 . The polynucleotide of claim 15 , wherein the second leader sequence comprises the amino acid sequence of SEQ ID NO: 15, SEQ ID NO: 44, or SEQ ID NO: 46, or an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 15, SEQ ID NO: 44, or SEQ ID NO: 46; or the nucleotide encoding the second leader sequence comprises the nucleotide sequence of SEQ ID NO: 16, SEQ ID NO: 45, or SEQ ID NO: 47, or a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 16, SEQ ID NO: 45, or SEQ ID NO: 47.
17 . The polynucleotide of claim 2 , wherein the first polypeptide comprises the amino acid sequence of SEQ ID NO: 17, or an amino acid sequence having at least 80% sequence identity thereof; or the sequence encoding the first polypeptide comprises the nucleotide sequence of SEQ ID NO: 18, or a nucleotide sequence having at least 80% sequence identity thereof.
18 . The polynucleotide of claim 2 , wherein the second polypeptide comprises the amino acid sequence of SEQ ID NO: 19, or an amino acid sequence having at least 80% sequence identity thereof; or the sequence encoding the second polypeptide comprises the nucleotide sequence of SEQ ID NO: 20, or a nucleotide sequence having at least 80% sequence identity thereof.
19 . The polynucleotide of claim 2 , wherein the sequence encoding the first polypeptide is operably linked to the sequence encoding a second polypeptide via a sequence encoding a self-cleaving peptide or an internal ribosomal entry site (IRES).
20 . The polynucleotide of claim 19 , wherein the self-cleaving peptide comprises the amino acid sequence of SEQ ID NO: 21, or an amino acid sequence having at least 80% sequence identity thereof; or the sequence encoding the self-cleaving peptide comprises the nucleotide sequence of SEQ ID NO: 22, or a nucleotide sequence having at least 80% sequence identity thereof.
21 . The polynucleotide of claim 1 , wherein the chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 23, or an amino acid sequence having at least 80% sequence identity thereof; or the polynucleotide comprises the nucleotide sequence of SEQ ID NO: 24, or a nucleotide sequence having at least 80% sequence identity thereof.
22 . A recombinant vector comprising the polynucleotide of claim 1 .
23 . A chimeric cytokine receptor encoded by the polynucleotide of claim 1 .
24 . A chimeric cytokine receptor, comprising two polypeptides, each comprising an extracellular domain of granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor, or a functional portion thereof, a transmembrane domain, and an intracellular domain of interleukin-18 receptor (IL-18) receptor, or a functional portion thereof, wherein the chimeric cytokine receptor improves effector function when expressed in an immune cell expressing a chimeric antigen receptor (CAR), an antigen specific T cell receptor (TCR) and/or bispecific antibody, relative to an immune cell not expressing the chimeric cytokine receptor.
25 . An isolated host cell comprising the polynucleotide of claim 1 .
26 . A pharmaceutical composition comprising the host cell of claim 25 and a pharmaceutically acceptable carrier and/or excipient.
27 . The polynucleotide of claim 1 , wherein the immune cell is a T cell or NK cell.
28 . The polynucleotide of claim 1 , wherein the chimeric cytokine receptor comprises
(i) a first polypeptide comprising an extracellular region of GM-CSF receptor α chain, or a functional portion thereof, a first transmembrane region, and an intracellular region of IL-18 receptor β chain, or a functional portion thereof; and
(ii) a second polypeptide comprising an extracellular region of GM-CSF receptor β chain, or a functional portion thereof, a second transmembrane region, and an intracellular region of IL-18 receptor α chain, or a functional portion thereof.
29 . The polynucleotide of claim 28 , wherein the extracellular region of GM-CSF receptor α chain comprises the amino acid sequence of SEQ ID NO: 1, or an amino acid sequence having at least 80% sequence identity thereof; or the sequence encoding the extracellular region of GM-CSF receptor α chain comprises the nucleotide sequence of SEQ ID NO: 2, or a nucleotide sequence having at least 80% sequence identity thereof.
30 . The polynucleotide of claim 28 , wherein the intracellular region of IL-18 receptor α chain comprises the amino acid sequence of SEQ ID NO: 5, or an amino acid sequence having at least 80% sequence identity thereof; or the sequence encoding the intracellular region of IL-18 receptor α chain comprises the nucleotide sequence of SEQ ID NO: 6, or a nucleotide sequence having at least 80% sequence identity thereof.
31 . The polynucleotide of claim 28 , wherein the extracellular region of GM-CSF receptor β chain comprises the amino acid sequence of SEQ ID NO: 7, or an amino acid sequence having at least 80% sequence identity thereof; or the sequence encoding the extracellular region of GM-CSF receptor β chain comprises the nucleotide sequence of SEQ ID NO: 8, or a nucleotide sequence having at least 80% sequence identity thereof.
32 . The polynucleotide of claim 28 , wherein the intracellular region of IL-18 receptor β chain comprises the amino acid sequence of SEQ ID NO: 11, or an amino acid sequence having at least 80% sequence identity thereof; or the sequence encoding the intracellular region of IL-18 receptor β chain comprises the nucleotide sequence of SEQ ID NO: 12, or a nucleotide sequence having at least 80% sequence identity thereof.
33 . The polynucleotide of claim 28 , wherein at least one of the transmembrane regions is derived from a transmembrane domain of IL-18 receptor or GM-CSF receptor.
34 . The polynucleotide of claim 28 , wherein the first transmembrane region comprises a transmembrane region of IL-18 receptor α chain.
35 . The polynucleotide of claim 34 , wherein the transmembrane region of IL-18 receptor α chain comprises the amino acid sequence of SEQ ID NO: 3, or an amino acid sequence having at least 80% sequence identity thereof; or the sequence encoding the transmembrane region of IL-18 receptor α chain comprises the nucleotide sequence of SEQ ID NO: 4, or a nucleotide sequence having at least 80% sequence identity thereof.
36 . The polynucleotide of claim 28 , wherein the second transmembrane region comprises a transmembrane region of IL-18 receptor β chain.
37 . The polynucleotide of claim 36 , wherein the transmembrane region of IL-18 receptor β chain comprises the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence having at least 80% sequence identity thereof; or the sequence encoding the transmembrane region of IL-18 receptor β chain comprises the nucleotide sequence of SEQ ID NO: 10, or a nucleotide sequence having at least 80% sequence identity thereof.
38 . The polynucleotide of claim 28 , wherein the first polypeptide further comprises a first leader sequence and/or the second polypeptide further comprises a second leader sequence.
39 . The polynucleotide of claim 38 , wherein the first leader sequence is derived from a leader sequence of GM-CSF receptor α chain.
40 . The polynucleotide of claim 38 , wherein the first leader sequence comprises the amino acid sequence of SEQ ID NO: 13, SEQ ID NO: 44, or SEQ ID NO: 46, or an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 13, SEQ ID NO: 44, or SEQ ID NO: 46; or the sequence encoding the first leader sequence comprises the nucleotide sequence of SEQ ID NO: 14, SEQ ID NO: 45, or SEQ ID NO: 47, or a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 14, SEQ ID NO: 45, or SEQ ID NO: 47.
41 . The polynucleotide of claim 38 , wherein the second leader sequence is derived from a leader sequence of GM-CSF receptor β chain.
42 . The polynucleotide of claim 41 , wherein the second leader sequence comprises the amino acid sequence of SEQ ID NO: 15, SEQ ID NO: 44, or SEQ ID NO: 46, or an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 15, SEQ ID NO: 44, or SEQ ID NO: 46; or the nucleotide encoding the second leader sequence comprises the nucleotide sequence of SEQ ID NO: 16, SEQ ID NO: 45, or SEQ ID NO: 47, or a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 16, SEQ ID NO: 45, or SEQ ID NO: 47.
43 . The polynucleotide of claim 28 , wherein the sequence encoding the first polypeptide is operably linked to the sequence encoding a second polypeptide via a sequence encoding a self-cleaving peptide or an internal ribosomal entry site (IRES).
44 . The polynucleotide of claim 43 , wherein the self-cleaving peptide comprises the amino acid sequence of SEQ ID NO: 21, or an amino acid sequence having at least 80% sequence identity thereof; or the sequence encoding the self-cleaving peptide comprises the nucleotide sequence of SEQ ID NO: 22, or a nucleotide sequence having at least 80% sequence identity thereof.