CHIMERIC ANTIGEN RECEPTOR AND NATURAL KILLER CELLS EXPRESSING SAME
The present invention provides a chimeric antigen receptor and natural killer cells expressing the same, and particularly, a chimeric antigen receptor (CAR) which includes an intracellular signaling domain including the whole or a portion of an OX40 ligand (CD252), thereby having excellent effects of increasing anticancer activity of immune cells, and immune cells expressing the same.
1 - 30 . (canceled)
31 . A chimeric antigen receptor, comprising:
an intracellular signaling domain which includes the whole or a portion of OX40 ligand (CD252).
32 . The chimeric antigen receptor according to claim 31 , further comprising:
a transmembrane domain linked to the intracellular signaling domain;
a spacer domain linked to the transmembrane domain; and
an extracellular domain linked to the spacer domain.
33 . The chimeric antigen receptor according to claim 32 , further comprising a signal sequence linked to the extracellular domain.
34 . The chimeric antigen receptor according to claim 33 , wherein the signal sequence includes the whole or a portion of CD16 and the signal sequence includes the whole or a portion of CD8α, respectively.
35 . The chimeric antigen receptor according to claim 32 , wherein the extracellular domain includes the whole or a portion of any one selected from the group consisting of an antigen-binding fragment of an antibody, a Fc receptor, a natural cytotoxicity receptor, NKG2D, 2B4 and DNAM-1.
36 . The chimeric antigen receptor according to claim 35 , wherein the antigen-binding fragment is Fab fragment, F(ab′) fragment, F(ab′)2 fragment or Fv fragment.
37 . The chimeric antigen receptor according to claim 36 , wherein the Fv fragment is a single-chain variable fragment (ScFv).
38 . The chimeric antigen receptor according to claim 35 , wherein the Fc receptor is selected from the group consisting of CD16, CD32, CD64, CD23, CD89 and variants thereof.
39 . The chimeric antigen receptor according to claim 38 , wherein the Fc receptor is CD16 or variants thereof.
40 . The chimeric antigen receptor according to claim 35 , wherein the natural cytotoxicity receptor is selected from the group consisting of NKp46, NKp30, NKp44, NKp80 and NKp65 receptors.
41 . The chimeric antigen receptor according to claim 32 , wherein the spacer domain includes the whole or a portion of any one selected from the group consisting of CD8α and CD28.
42 . The chimeric antigen receptor according to claim 32 , wherein the transmembrane domain includes the whole or a portion of any one selected from the group consisting of CD8α and CD28.
43 . The chimeric antigen receptor according to claim 31 , wherein the intracellular signaling domain further includes the whole or a portion of CD3-zeta.
44 . The chimeric antigen receptor according to claim 43 , wherein the whole or a portion of CD3-zeta and the whole or a portion of OX40 ligand are arranged in order from a cell membrane toward an inside of the cell.
45 . The chimeric antigen receptor according to claim 32 , further comprising an AAA linker between the extracellular domain and the spacer domain.
46 . A chimeric antigen receptor, comprising intracellular signaling domains which comprise:
a first intracellular signaling domain including the whole or a portion of any one selected from the group consisting of CD28 and 4-1BB;
a second intracellular signaling domain including the whole or a portion of any one selected from the group consisting of OX40 ligand, OX40 and 4-1BB; and
a third intracellular signaling domain including the whole or a portion of CD3-zeta,
wherein the first, second and third intracellular signaling domains are arranged in order from a cell membrane toward an inside of the cell.
47 . The chimeric antigen receptor according to claim 46 , further comprising:
a transmembrane domain linked to the intracellular signaling domain;
a spacer domain linked to the transmembrane domain; and
an extracellular domain linked to the spacer domain.
48 . The chimeric antigen receptor according to claim 47 , further comprising a signal sequence linked to the extracellular domain.
49 . The chimeric antigen receptor according to claim 48 , wherein the signal sequence includes the whole or a portion of CD16 and the signal sequence includes the whole or a portion of CD8α, respectively.
50 . The chimeric antigen receptor according to claim 47 , wherein the extracellular domain includes the whole or a portion of any one selected from the group consisting of an antigen-binding fragment of an antibody, a Fc receptor, a natural cytotoxicity receptor, NKG2D, 2B4 and DNAM-1.
51 . The chimeric antigen receptor according to claim 47 , wherein the spacer domain includes the whole or a portion of any one selected from the group consisting of CD8α and CD28.
52 . The chimeric antigen receptor according to claim 47 , wherein the transmembrane domain includes the whole or a portion of any one selected from the group consisting of CD8α and CD28.
53 . The chimeric antigen receptor according to claim 46 , wherein the first intracellular signaling domain includes the whole or a portion of CD28;
the second intracellular signaling domain includes the whole or a portion of the OX40 ligand; and
the third intracellular signaling domain includes the whole or a portion of CD3-zeta.
54 . The chimeric antigen receptor according to claim 47 , further comprising an AAA linker between the extracellular domain and the spacer domain.
55 . An immune cell expressing the chimeric antigen receptor according to claim 31 .
56 . The immune cell according to claim 55 , wherein the immune cell is a natural killer cell (NK cell).
57 . A pharmaceutical composition for treatment of tumor, comprising the immune cell according to claim 55 as an active ingredient.
58 . The pharmaceutical composition according to claim 57 , further comprising an antibody as an active ingredient when the extracellular domain is a Fc receptor.
59 . A nucleic acid sequence encoding the chimeric antigen receptor according to claim 31 .
60 . The nucleic acid sequence according to claim 59 , wherein the nucleic acid sequence encodes one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 33, 41, 43, 45, 47, 49, 51, 53, 55, 69, 71, 77, 81, 83, 85, 87, 89, 91 and 93, or variants thereof having a sequence identity of 80% or more.
61 . The nucleic acid sequence according to claim 60 , wherein the nucleic acid sequence includes one or more nucleic acid sequence selected from the group consisting of SEQ ID NOs: 32, 40, 42, 44, 46, 48, 50, 52, 54, 68, 70, 76, 80, 82, 84, 86, 88, 90 and 92, or variants thereof having a sequence identity of 80% or more.
62 . A method for treatment of tumor, comprising administering the immune cell according to claim 55 to a subject.