MULTI-CHAIN CHIMERIC POLYPEPTIDES AND USES THEREOF
Provided herein are multi-chain chimeric polypeptides and uses thereof for treating cancer, aging related diseases or conditions, and/or an infectious disease. Further provided, are methods for stimulating an immune cell, inducing or increasing proliferation of an immune cell, and/or inducing differentitation of an immune cell into a memory-like immune cell by contacting the immune cell with the multi-chain chimeric polypeptides disclosed herein.
1 . A multi-chain chimeric polypeptide comprises:
(a) a first chimeric polypeptide comprising:
(i) a first target-binding domain comprising:
a first sequence that is at least 90% identical to SEQ ID NO:2, wherein the amino acid at position 32 in SEQ ID NO:2 is asparagine and the amino acid at position 119 in SEQ ID NO:2 is alanine; and
a second sequence that is at least 90% identical to SEQ ID NO:2, wherein the amino acid at position 32 in SEQ ID NO:2 is asparagine and the amino acid at position 119 in SEQ ID NO:2 is alanine;
(ii) a soluble tissue factor domain comprising a sequence that is at least 90% identical to SEQ ID NO:1; and
(iii) a first domain of a pair of affinity domains comprising a sequence that is at least 90% identical to SEQ ID NO:13; and
(b) a second chimeric polypeptide comprising:
(i) a second domain of a pair of affinity domains comprising a sequence that is at least 90% identical to SEQ ID NO:11; and
(ii) a second target-binding domain comprising:
a first sequence that is at least 90% identical to SEQ ID NO:2, wherein the amino acid at position 32 in SEQ ID NO:2 is asparagine and the amino acid at position 119 in SEQ ID NO:2 is alanine; and
a second sequence that is at least 90% identical to SEQ ID NO:2, wherein the amino acid at position 32 in SEQ ID NO:2 is asparagine and the amino acid at position 119 in SEQ ID NO:2 is alanine,
wherein:
the first chimeric polypeptide and the second chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains.
2 . The multi-chain chimeric polypeptide of claim 1 , wherein the first target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 4.
3 . The multi-chain chimeric polypeptide of claim 1 , wherein the first chimeric polypeptide comprises a sequence that is at least 80% identical to SEQ ID NO:6.
4 . The multi-chain chimeric polypeptide of claim 1 , wherein the first chimeric polypeptide comprises a sequence of SEQ ID NO: 7.
5 . The multi-chain chimeric polypeptide of claim 1 , wherein the second target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO:4.
6 . The multi-chain chimeric polypeptide of claim 1 , wherein the second chimeric polypeptide comprises a sequence that is at least 80% identical to SEQ ID NO:5.
7 . The multi-chain chimeric polypeptide of claim 1 , wherein the second chimeric polypeptide comprises a sequence of SEQ ID NO:8.
8 . A pharmaceutical composition comprising the multi-chain chimeric polypeptide of claim 1 .
9 . A kit comprising at least one dose of the composition of claim 8 .
10 . A method of stimulating an immune cell, and/or inducing or increasing proliferation of an immune cell, the method comprising contacting an immune cell with an effective amount of the multi-chain chimeric polypeptide of claim 1 .
11 . The method of claim 10 , wherein the immune cell is contacted in vitro or is contacted in vivo.
12 . The method of claim 10 , wherein the immune cell is selected from the group consisting of: an immature thymocyte, a peripheral blood lymphocyte, a naïve T cell, a pluripotent Th cell precursor, a lymphoid progenitor cell, a Treg cell, a Th17 cell, a Th22 cell, a Th9 cell, a Th2 cell, a Th1 cell, a Th3 cell, γδ T cell, an αβ T cell, a tumor-infiltrating T cell, a CD8+ T cell, a CD4+ T cell, a natural killer T cell, a mast cell, a macrophage, a neutrophil, a dendritic cell, a basophil, an eosinophil, and a natural killer cell.
13 . The method of 10 , wherein the immune cell has previously been genetically modified to express a chimeric antigen receptor or a recombinant T-cell receptor.
14 . A method of inducing differentiation of an immune cell into a memory or memory-like immune cell, the method comprising contacting an immune cell with an effective amount of the multi-chain chimeric polypeptide of claim 1 .
15 . The method of claim 14 , wherein the immune cell is contacted in vitro or is contacted in vivo.
16 . The method of claim 14 , wherein the immune cell is selected from the group consisting of: an immature thymocyte, a peripheral blood lymphocyte, a naïve T cell, a pluripotent Th cell precursor, a lymphoid progenitor cell, a Treg cell, a Th17 cell, a Th22 cell, a Th9 cell, a Th2 cell, a Th1 cell, a Th3 cell, γδ T cell, an αβ T cell, a tumor-infiltrating T cell, a CD8+ T cell, a CD4+ T cell, a natural killer T cell, a mast cell, a macrophage, a neutrophil, a dendritic cell, a basophil, an eosinophil, and a natural killer cell.
17 . The method of claim 14 , wherein the immune cell has previously been genetically modified to express a chimeric antigen receptor or a recombinant T-cell receptor.
18 . A method of killing a cancer cell, an infected cell, or a senescent cell in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the multi-chain chimeric polypeptide of claim 1 .
19 . The method of claim 18 , wherein the subject has been identified or diagnosed as having a cancer, an aging-related disease or condition, or an infectious disease.
20 . A method of treating a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the pharmaceutical composition of claim 8 , wherein the subject has been identified or diagnosed as having a cancer, an aging-related disease or condition, or an infectious disease.
21 . A nucleic acid encoding any of the multi-chain chimeric polypeptide of claim 1 .
22 . A vector comprising the nucleic acid of claim 21 .
23 . A cell comprising the nucleic acid of claim 21 .
24 . A cell comprising the vector of claim 22 .
25 . A method of producing a multi-chain chimeric polypeptide, the method comprising:
culturing the cell of claim 23 in a culture medium under conditions sufficient to result in the production of the multi-chain chimeric polypeptide; and
recovering the multi-chain chimeric polypeptide from the cell and/or the culture medium.
26 . A multi-chain chimeric polypeptide produced by the method of claim 25 .