USE OF IL-7R-ALPHA-GAMMA AGONIST PEPTIDES IN CELL MANUFACTURING
The use of IL-7Rαγc agonist peptides in cell manufacturing is disclosed. The IL-7Rαγc agonist peptides can facilitate the selective growth of immune cell populations. Culture media comprising the IL-7Rαγc agonist peptides, methods of expanding a target immune cell population, and methods of manufacturing a target immune cell population are disclosed. The enriched immune cell populations can be used in immunotherapy.
1 . A culture medium for expanding a target immune cell population comprising a first stimulant of proliferation for the target immune cell population proliferation, wherein the first stimulant comprises an IL-7Rαγc agonist peptide, wherein the IL-7Rαγc agonist peptide comprises:
(a) an IL-7Rα ligand, wherein the IL-7Rα ligand comprises:
an amino acid sequence of any one of SEQ ID NO: 43-46, 100-117, 118-193, 197-245 and 53-60;
a truncated amino acid sequence of any one of SEQ ID NO: 43-46, 100-117, 118-193, 197-245 and 53-60;
an amino acid sequence of any one of SEQ ID NO: 43-46, 100-117, 118-193, 197-245 and 53-606 having from 1 to 5 amino acid substitutions;
an amino acid sequence of any one of SEQ ID NO: 43-46, 100-117, 118-193, 197-245 and 53-60 having from 1 to 5 flanking glycines on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus;
an amino acid sequence having greater than 60% sequence similarity to an amino acid sequence of any one of SEQ ID NO: 43-46, 100-117, 118-193, 197-245 and 53-60; or
a combination of any of the foregoing; and
(b) an Rγc ligand, wherein the Rγc ligand comprises:
an amino acid sequence of any one of SEQ ID NO: 246-327 and 47-52;
a truncated amino acid sequence of any one of SEQ ID NO: 246-327 and 47-52;
an amino acid sequence of any one of SEQ ID NO: 246-327 and 47-52 having from 1 to 5 amino acid substitutions;
an amino acid sequence of any one of SEQ ID NO: 246-327 and 47-52 having from 1 to 5 flanking glycines on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus;
an amino acid sequence having greater than 60% sequence similarity to an amino acid sequence of any one of SEQ ID NO: 246-327 and 47-52; or
a combination of any of the foregoing.
2 . The culture medium of claim 1 , wherein the IL-7Rαγc agonist peptide comprises:
an IL-7Rα ligand comprising an amino acid sequence of any one of SEQ ID NO: 100-117, 118-193, 197-245 and 53-60; and
an Rγc ligand comprising an amino acid sequence of any one of SEQ ID NO: 246-327 and 47-52.
3 . The culture medium of claim 1 , wherein the IL-7Rα ligand comprises an amino acid sequence of any one of SEQ ID NO: 114 and 152.
4 . The culture medium of claim 1 , wherein the Rγc ligand comprises an amino acid sequence of any one of SEQ ID NO: 261 and 325-327.
5 . The culture medium of claim 1 , wherein the IL-7Rαγc agonist peptide comprises:
an amino acid sequence of any one of SEQ ID NO: 443-454 and 368-442;
a truncated amino acid sequence of any one of SEQ ID NO: 443-454 and 368-442;
an amino acid sequence of any one of SEQ ID NO: 443-454 and 368-442 having from 1 to 5 amino acid substitutions;
an amino acid sequence of any one of SEQ ID NO: 443-454 and 368-442 having from 1 to 5 flanking glycines on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus;
an amino acid sequence having greater than 60% sequence similarity to an amino acid sequence of any one of SEQ ID NO: 443-454 and 368-442; or
a combination of any of the foregoing.
6 . The culture medium of claim 1 , wherein the IL-7Rαγc agonist peptide comprises an amino acid sequence of any one of SEQ ID NO: 394-404.
7 . The culture medium any one of aspects claim 1 , wherein the culture medium comprises a second stimulant of proliferation for the target immune cell population.
8 . The culture medium of claim 7 , wherein the second stimulant comprises an IL-2Rβγc agonist peptide.
9 . The culture medium of claim 8 , wherein the IL-2Rβγc agonist peptide comprises an amino acid sequence of any one of SEQ ID NO: 359-366, 61-76, and 77-99.
10 . The culture medium of claim 9 , wherein the IL-2Rβγc agonist peptide comprises an amino acid sequence of any one of SEQ ID NO: 91-99 and 359.
11 . The culture medium of claim 7 , wherein the second stimulant comprises an IL-15Rβγc agonist peptide.
12 . The culture medium of claim 1 , wherein the culture medium comprises an antibody directed to the target immune cell population.
13 . A method of expanding a target immune cell population of an initial immune cell population, comprising incubating an initial immune cell population in the culture medium of claim 1 to provide an expanded target immune cell population.
14 . A method of manufacturing an immune cell population, comprising activating a target immune cell population of a population of primary cells to provide an activated target immune cell population; and
expanding the activated target immune cell population, wherein expanding comprises incubating the activated target immune cell population in the presence of an IL-7Rαγc agonist peptide to provide an expanded target immune cell population, wherein the IL-7Rαγc agonist peptide comprises:
(a) an IL-7Rα ligand, wherein the IL-7Rα ligand comprises:
an amino acid sequence of any one of SEQ ID NO: 43-46, 100-117, 118-193, 197-245 and 53-60;
a truncated amino acid sequence of any one of SEQ ID NO: 43-46, 100-117, 118-193, 197-245 and 53-60;
an amino acid sequence of any one of SEQ ID NO: 43-46, 100-117, 118-193, 197-245 and 53-60 having from 1 to 5 amino acid substitutions;
an amino acid sequence of any one of SEQ ID NO: 43-46, 100-117, 118-193, 197-245 and 53-60 having from 1 to 5 flanking glycines on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus;
an amino acid sequence having greater than 60% sequence similarity to an amino acid sequence of any one of SEQ ID NO: 43-46, 100-117, 118-193, 197-245 and 53-60; or
a combination of any of the foregoing; and
(b) an Rγc ligand, wherein the Rγc ligand comprises:
an amino acid sequence of any one of SEQ ID NO: 246-327 and 47-52;
a truncated amino acid sequence of any one of SEQ ID NO: 246-327 and 47-52;
an amino acid sequence of any one of SEQ ID NO: 246-327 and 47-52 having from 1 to 5 amino acid substitutions;
an amino acid sequence of any one of SEQ ID NO: 246-327 and 47-52 having from 1 to 5 flanking glycines on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus;
an amino acid sequence having greater than 60% sequence similarity to an amino acid sequence of any one of SEQ ID NO: 246-327 and 47-52; or
a combination of any of the foregoing.
15 . An enriched population of immune cells prepared using the culture medium of claim 1 .
16 . An enriched population of immune cells prepared using the method of claim 13 .
17 . An enriched population of immune cells prepared using the method of claim 14 .
18 . A pharmaceutical composition comprising the enriched population of claim 16 .
19 . A method of treating a disease in a patient comprising administering to a patient in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 18 .
20 . A pharmaceutical composition comprising the enriched population of claim 17 .
21 . A method of treating a disease in a patient comprising administering to a patient in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 20 .
22 . An immobilized IL-7Rαγc agonist peptide, comprising an IL-7Rαγc agonist peptide bound to a substrate, wherein the IL-7Rαγc agonist peptide comprises:
(a) an IL-7Rα ligand, wherein the IL-7Rα ligand comprises:
an amino acid sequence of any one of SEQ ID NO: 43-46, 100-117, 118-193, 197-245 and 53-60;
a truncated amino acid sequence of any one of SEQ ID NO: 43-46, 100-117, 118-193, 197-245 and 53-60;
an amino acid sequence of any one of SEQ ID NO: 43-46, 100-117, 118-193, 197-245 and 53-60 having from 1 to 5 amino acid substitutions;
an amino acid sequence of any one of SEQ ID NO: 43-46, 100-117, 118-193, 197-245 and 53-60 having from 1 to 5 flanking glycines on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus;
an amino acid sequence having greater than 60% sequence similarity to an amino acid sequence of any one of SEQ ID NO: 43-46, 100-117, 118-193, 197-245 and 53-60; or
a combination of any of the foregoing; and
(b) an Rγc ligand, wherein the Rγc ligand comprises:
an amino acid sequence of any one of SEQ ID NO: 246-327 and 47-52;
a truncated amino acid sequence of any one of SEQ ID NO: 246-327 and 47-52;
an amino acid sequence of any one of SEQ ID NO: 246-327 and 47-52 having from 1 to 5 amino acid substitutions;
an amino acid sequence of any one of SEQ ID NO: 246-327 and 47-52 having from 1 to 5 flanking glycines on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus;
an amino acid sequence having greater than 60% sequence similarity to an amino acid sequence of any one of SEQ ID NO: 246-327 and 47-52; or
a combination of any of the foregoing.