IMMUNE EFFECTOR CELL AND USE THEREOF
A geneticall engineered cell. The cell expresses an exogenous receptor binding to an antigen, and expresses increased RUNX3 or exogenous RUNX3. Also provided are a use of the cell and a method for treating tumors.
1 . A genetically engineered cell, wherein the cell expresses an exogenous receptor binding to an antigen and expresses an increased level of RUNX3 or exogenous RUNX3.
2 . The cell of claim 1 , wherein the cell is an immune effector cell;
preferably the immune effector cell is a T cell.
3 . (canceled)
4 . The cell of claim 1 , wherein the RUNX3 is a full-length human RUNX3 or a fragment of human RUNX3 having the same function as the full-length human RUNX3;
preferably the RUNX3 is at least 90% identical to the sequence as shown in SEQ ID NO: 20.
5 . (canceled)
6 . The cell of claim 1 , wherein the RUNX3 is constitutively expressed; or the RUNX3 is inducibly expressed.
7 . (canceled)
8 . The cell of claim 1 , wherein the antigen is a tumor antigen or a pathogen antigen, preferably a tumor antigen;
preferably the tumor antigen is a solid tumor antigen;
more preferably the tumor antigen is GPC3 or claudin 18.2.
9 - 10 . (canceled)
11 . The cell of claim 1 , wherein the receptor is a chimeric receptor selected from the group consisting of chimeric antigen receptor (CAR), modified T cell (antigen) receptor (TCR), T cell fusion protein (TFP), T Cell antigen coupler (TAC) or a combination thereof;
preferably, the receptor is a chimeric antigen receptor;
more preferably the intracellular domain of the chimeric antigen receptor comprises the intracellular costimulatory signaling domain of CD137.
12 - 13 . (canceled)
14 . The cell of claim 1 , wherein the extracellular domain of the receptor has an amino acid sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 42 or SEQ ID NO: 22.
15 . The cell of claim 1 , wherein the intracellular domain of the receptor contains has an amino acid sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 47;
preferably the intracellular domain of the receptor also has an amino acid sequence that is at least 90% identical to SEQ ID NO: 46 or SEQ ID NO: 49, or comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 46 or SEQ ID NO: 49.
16 . (canceled)
17 . The cell of claim 11 , wherein the cell comprises a nucleic acid sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:
17, 19, 53, 54, 55, or 56.
18 . The cell of claim 1 , wherein the cell also expresses an exogenous cytokine receptor-binding protein, or an exogenous cytokine or a polypeptide thereof.
19 . The cell of claim 18 , wherein the cell also expresses an exogenous cytokine; or
the exogenous cytokine receptor-binding protein can specifically bind to the corresponding cytokine receptor and enhance activities of the receptor; or
the exogenous cytokine or polypeptide thereof can specifically bind to the corresponding cytokine receptor and enhance activities of the receptor.
20 - 21 . (canceled)
22 . The cell of claim 19 , wherein the cytokine has an amino acid sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the sequence as shown in SEQ ID NO: 39, 41 or 36.
23 . The cell of claim 1 , wherein the expression level of RUNX3 is increased by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% as compared with the wild type; or
the increase in the expression level of the RUNX3 as compared with the wild type is sufficient to increase the retention of the cell in non-lymphatic sites.
24 . (canceled)
25 . The cell of claim 19 , wherein the cytokine is constitutively expressed; or
the cytokine is inducibly expressed.
26 . (canceled)
27 . The cell of claim 1 , wherein the cell expresses an exogenous RUNX3;
preferably the receptor and/or RUNX3 are expressed by using a viral vector; and
preferably, the viral vector includes: a lentiviral vector, retroviral vector or adenoviral vector.
28 . (canceled)
29 . An expression construct, wherein the expression construct comprises sequentially connected expression cassette 1 of an antigen-binding receptor and expression cassette 2 of RUNX3, wherein the expression cassettes are optionally connected by tandem fragments selected from the group consisting of F2A, PA2, T2A, and E2A;
preferably the expression cassette of the antigen-binding receptor comprises a nucleic acid sequence encoding the sequence as shown in SEQ ID NO: 57, 58, 59, 60, 61, or 62, and the expression cassette of RUNX3 has a nucleic acid sequence encoding the sequence as shown in SEQ ID NO: 20; or
the expression construct further comprises expression cassette 3, the expression cassette 3 comprises a nucleic acid sequence encoding a cytokine, and the sequence of the cytokine is an amino acid sequence which is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the sequence as shown in SEQ ID NO: 36, 39, or 41.
30 - 31 . (canceled)
32 . An expression vector or a virus, wherein the expression vector comprises the expression construct of claim 29 , and the virus comprises the expression vector.
33 . (canceled)
34 . A method for increasing the viability of an immune effector cell expressing a chimeric receptor in an individual wherein the expression level of RUNX3 in the immune effector cell is increased, and preferably, the RUNX3 is constitutively expressed or inducibly expressed; or
the expression level of RUNX3 in the immunity effector cell is increased by expressing exogenous RUNX3 in the immune effector cell;
preferably, the RUNX3 is a full-length human RUNX3 or a fragment of human RUNX3 having the same function as the full-length human RUNX3;
more preferably, the RUNX3 has at least 90% identity with the sequence as shown in SEQ ID NO: 20.
35 - 39 . (canceled)
40 . The method of claim 34 , wherein the immune effector cell also co-expresses a cytokine; or
the individual is also administered with a cytokine; or
the individual is also administered with a chemotherapeutic drug.
41 . (canceled)
42 . The method of claim 34 , wherein the immune effector cell is a T cell; and/or
the chimeric receptor is a chimeric antigen receptor.
43 - 44 . (canceled)
45 . A method for inhibiting a tumor or inhibiting a pathogen, comprising a step of administering a therapeutically sufficient amount of the cell of claim 1 to a subject in need thereof. cm 46 . A pharmaceutical composition for inhibiting a tumor or inhibiting a pathogen, wherein the pharmaceutical composition comprises the cell of claim 1 and a pharmaceutically acceptable carrier or excipient.
47 - 52 . (canceled)