Compositions and Methods for Modulating the Immune System
Embodiments provided for herein relate to compositions and methods related to chimeric proteins that can be used to modulate a subject's immune system and, for example, treat cancer.
1 . A protein comprising:
an extracellular domain of PD-1;
a transmembrane domain selected from the group consisting of: 4-1 BB transmembrane domain, CD28 transmembrane domain, CD27 transmembrane domain, and ICOS transmembrane domain; and
an intracellular signaling domain selected from the group consisting of 4-1 BB intracellular signaling domain, CD28 intracellular signaling domain, CD27 intracellular signaling domain, and ICOS intracellular signaling domain, and any combination thereof.
2 . The protein of claim 1 , wherein the transmembrane domain is the 4-1BB transmembrane domain.
3 . The protein of claim 2 , wherein the intracellular signaling domain is selected from the group consisting of: 4-1 BB intracellular signaling domain, CD28 intracellular signaling domain, CD27 intracellular signaling domain, and ICOS intracellular signaling domain.
4 . The protein of claim 2 , wherein the intracellular signaling domain is the intracellular domain of 4-1 BB and an intracellular domain selected from the group consisting of: CD28 intracellular signaling domain, CD27 intracellular signaling domain, and ICOS intracellular signaling domain.
5 . The protein of claim 1 , wherein the transmembrane domain is the CD28 transmembrane domain.
6 . The protein of claim 5 , wherein the intracellular signaling domain is selected from the group consisting of: 4-1 BB transmembrane domain, CD28 transmembrane domain, CD27 transmembrane domain, and ICOS transmembrane domain.
7 . The protein of claim 5 , wherein the intracellular signaling domain is the intracellular domain of 4-1 BB and an intracellular domain selected from the group consisting of: CD28 intracellular signaling domain, CD27 intracellular signaling domain, and ICOS intracellular signaling domain.
8 . The protein of claim 1 , wherein the transmembrane domain is the CD27 transmembrane domain.
9 . The protein of claim 8 , wherein the intracellular signaling domain is selected from the group consisting of: 4-1 BB intracellular signaling domain, CD28 intracellular signaling domain, CD27 intracellular signaling domain, and ICOS intracellular signaling domain.
10 . The protein of claim 8 , wherein the intracellular signaling domain is the intracellular domain of 4-1 BB and an intracellular domain selected from the group consisting of: CD28 intracellular signaling domain, CD27 intracellular signaling domain, and ICOS intracellular signaling domain.
11 . The protein of claim 1 , wherein the transmembrane domain is the ICOS transmembrane domain.
12 . The protein of claim 11 , wherein the intracellular signaling domain is selected from the group consisting of: 4-1 BB intracellular signaling domain, CD28 intracellular signaling domain, CD27 intracellular signaling domain, and ICOS intracellular signaling domain.
13 . The protein of claim 11 , wherein the intracellular signaling domain is the intracellular domain of 4-1 BB and the intracellular domain selected from the group consisting of: CD28 intracellular signaling domain, CD27 intracellular signaling domain, and ICOS intracellular signaling domain.
14 . The protein of claim 1 , wherein the protein comprises a sequence of SEQ ID NO: 21, 23, 25, 27, 29, or 31.
15 . The protein of claim 1 , wherein the protein comprises a leader sequence.
16 . The protein of claim 15 , wherein the protein comprises a CD8 leader sequence.
17 . The protein of claim 16 , wherein the CD8 leader sequence comprises SEQ ID NO: 1.
18 . A nucleic acid molecule encoding the protein of claim 1 .
19 . A nucleic acid molecule comprising a sequence of SEQ ID NO: 22, 24, 26, 28, 30, or 32.
20 . The nucleic acid molecule of claim 19 , further comprising a leader sequence encoded by SEQ ID NO: 2.
21 . A recombinant cell, comprising the nucleic acid molecule of claim 19 .
22 . A recombinant cell, comprising the nucleic acid molecule of claim 18 .
23 . A recombinant cell, comprising the protein of claim 1 .
24 . The cell of claim 21 , wherein the cell is a lymphocyte or a T cell.
25 . The cell of claim 21 , wherein the cell is a tumor infiltrating lymphocyte (“TIL”).
26 . The cell of claim 21 , wherein the cell is a marrow infiltrating lymphocyte (“MIL”).
27 . The cell of claim 26 , wherein the MIL is a hypoxic MIL.
28 . A method for making a recombinant cell, comprising transfecting or infecting a cell with a nucleic acid molecule encoding a protein of claim 1 .
29 . The method of claim 28 , wherein the cell is a MIL.
30 . The method of claim 28 further comprising incubating the MIL under hypoxic conditions prior to transfecting or infecting the cell with the nucleic acid molecule encoding the protein.
31 . The method of claim 30 , wherein the hypoxic conditions comprise about 0.5% to about 5% oxygen gas.
32 . The method of claim 30 , wherein the hypoxic conditions comprise about 1% to about 2% oxygen gas.
33 . The method of claim 30 , further comprising incubating the cells under normoxic conditions after the hypoxic incubation.
34 . The method of claim 28 , further comprising contacting the cell with anti-CD3/anti-CD28 beads.
35 . A method for increasing an immune response in a subject, comprising administering to the subject the recombinant cell of claim 21 .
36 . The method of claim 35 , further comprising making the recombinant cell, wherein making the recombinant cell comprises transfecting a cell with a nucleic acid encoding the protein.
37 . The method of claim 35 , further comprising isolating the cell from the subject.
38 . The method of claim 35 , wherein the subject has a neoplasm.
39 . The method of claim 35 , wherein the neoplasm is a leukemia, lymphoma, or multiple myeloma.
40 . The method of claim 35 , wherein the subject is a human.
41 . A method for treating a neoplasm in a subject, comprising administering to the subject the recombinant cell of claim 21 .
42 . The method of claim 41 , further comprising making the recombinant cell, wherein making the recombinant cell comprises transfecting a cell with a nucleic acid encoding the chimeric transmembrane protein.
43 . The method of claim 41 , further comprising isolating the cell from the subject.
44 . The method of claim 41 , wherein the subject has a neoplasm.
45 . The method of claim 44 , wherein the neoplasm is a multiple myeloma, leukemia or lymphoma.
46 . The method of claim 41 , wherein the subject is a human.
47 . The method of claim 41 , further comprising prior to administering the cell to the subject:
contacting the cell with anti-CD3/anti-CD28 beads;
incubating the cell under hypoxic conditions; and
incubating the cell under normoxic conditions.
48 . The method of claim 47 , wherein the cell is incubated under hypoxic conditions for about 0.5 to about 4 days.
49 . The method of claim 47 , wherein the cell is incubated under normoxic conditions for about 0.5 to about 4 days.