Chimeric antigen receptors targeting CD127 and use thereof
The presently disclosed subject matter provides methods for treating neoplasia using cells comprising an antigen-recognizing receptor (e.g., a chimeric antigen receptor (CAR)) that specifically targets CD127.
1 . A method for reducing tumor burden in a subject having a hematological tumor, treating and/or preventing a hematological tumor in a subject, and/or increasing or lengthening survival of a subject having a hematological tumor, comprising administering to the subject
a) an effective amount of cells comprising an antigen-recognizing receptor,
b) a pharmaceutical composition comprising an effective amount of cells comprising an antigen-recognizing receptor; or
c) a nucleic acid composition comprising a polynucleotide encoding an antigen-recognizing receptor;
wherein the antigen-recognizing receptor comprises a chimeric antigen receptor (CAR) comprising an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain, wherein the extracellular antigen-binding domain specifically binds CD127 comprises a heavy chain variable region (V H ) and a light chain variable region (V L ), which are positioned from a N- to a C-terminus: V H -V L , and
wherein the heavy chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 6, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 7, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 8, and the light chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 9, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 10, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11.
2 . Method of claim 1 , wherein the extracellular antigen-binding domain is a single-chain variable fragment (scFv), a Fab, or a F(ab) 2 .
3 . The method of claim 1 , wherein the extracellular antigen-binding domain is a humanized scFv.
4 . The method of claim 1 , wherein the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 12 and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 13.
5 . The method of claim 1 , wherein the extracellular antigen-binding domain comprises a linker between a heavy chain variable region and a light chain variable region.
6 . The method of claim 5 , wherein the linker consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4.
7 . The method of claim 1 , wherein the extracellular antigen-binding domain comprises the amino acid sequence set forth in SEQ ID NO: 14.
8 . The method of claim 1 ,
wherein the transmembrane domain comprises a CD8 polypeptide, a CD28 polypeptide, a CD32 polypeptide, a CD4 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a CTLA-4 polypeptide, a PD-1 polypeptide, a LAG-3 polypeptide, a 2B4 polypeptide, a BTLA polypeptide, a synthetic peptide (not based on a protein associated with the immune response), or a combination thereof; and/or
wherein the intracellular signaling domain comprises a CD35 polypeptide.
9 . The method of claim 8 , wherein the transmembrane domain comprises a CD28 polypeptide.
10 . The method of claim 1 , wherein the intracellular signaling domain further comprises at least one co-stimulatory signaling region.
11 . The method of claim 10 , wherein the at least one co-stimulatory signaling region comprises a CD28 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a DAP-10 polypeptide, or a combination thereof.
12 . The method of claim 1 , wherein the antigen-recognizing receptor comprises the amino acid sequence set forth in SEQ ID NO: 27.
13 . The method of claim 1 , wherein the antigen-recognizing receptor is recombinantly expressed and/or is expressed from a vector.
14 . The method of claim 1 , wherein the polynucleotide comprises or consists of the nucleotide acid sequence set forth in SEQ ID NO: 28.
15 . The method of claim 1 , wherein the nucleic acid composition is a vector.
16 . The method of claim 15 , wherein the vector is a γ-retroviral vector.
17 . The method of claim 1 , wherein the cell is transduced with the antigen-recognizing receptor.
18 . The method of claim 1 , wherein the cell is an immunoresponsive cells, a cell of the lymphoid lineage, or a cell of the myeloid lineage.
19 . The method of claim 1 , wherein the cell is selected from the group consisting of a T-cell, a Natural Killer (NK) cell, and a stem cell from which a lymphoid cell may be differentiated.
20 . The method of claim 1 , wherein
a) the T-cell is a cytotoxic T lymphocyte (CTL) or a regulatory T-cell; or
b) the stem cell is a pluripotent stem cell, an embryoid stem cell, or an induced pluripotent stem cell.
21 . The method of claim 1 , wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.
22 . The method of claim 1 , wherein the method reduces the number of tumor cells, reduces tumor size, and/or eradicates the tumor in the subject.
23 . The method of claim 1 , wherein the hematological tumor is selected from the group consisting of acute lymphoblastic leukemia (ALL), Hodgkin's lymphoma, non-Hodgkin's lymphoma, T-cell cutaneous lymphoma.
24 . The method of claim 23 , wherein the acute lymphoblastic leukemia (ALL) is associated with gain-mutation of the IL7-R/TSLP pathway.
25 . The method of claim 23 , wherein the acute lymphoblastic leukemia (ALL) is T-cell acute lymphoblastic leukemia (T-ALL) or B-cell acute lymphoblastic leukemia (B-ALL).
26 . The method of claim 1 , wherein the hematological tumor is T-cell acute lymphoblastic leukemia (T-ALL).