T cell receptors and methods of use thereof
The present disclosure is directed recombinant T cell receptors capable of binding a CCND1 epitope and nucleic acid molecules encoding the same. In some aspects, the nucleic acid molecules further comprise a second nucleotide sequence, wherein the second nucleotide sequence or the polypeptide encoded by the second nucleotide sequence inhibits the expression of an endogenous TCR. Other aspects of the disclosure are directed to vectors comprising the nucleic acid molecule and cells comprising the recombinant TCR, the nucleic acid molecule, or the vector. Still other aspects of the disclosure are directed to methods of using the same. In some aspects, the methods comprise treating a cancer in a subject in need thereof.
1 . A nucleic acid molecule comprising
(i) a first nucleotide sequence encoding a recombinant T cell receptor (TCR) or an antigen binding portion thereof that specifically binds human G1/S-specific cyclin-D1 (CCND1) (“anti-CCND1 TCR”); wherein the anti-CCND1 TCR comprises an alpha chain variable domain comprising an alpha chain CDR1, an alpha chain CDR2, and an alpha chain CDR3; and a beta chain variable domain comprising a beta chain CDR1, a beta chain CDR2, and a beta chain CDR3; and wherein:
(a) the beta chain CDR3 of the anti-CCND1 TCR comprises the amino acid sequence as set forth in SEQ ID NO: 10;
(b) the beta chain CDR2 of the anti-CCND1 TCR comprises the amino acid sequence as set forth in SEQ ID NO: 9;
(c) the beta chain CDR1 of the anti-CCND1 TCR comprises the amino acid sequence as set forth in SEQ ID NO: 8;
(d) the alpha chain CDR3 of the anti-CCND1 TCR comprises the amino acid sequence as set forth in SEQ ID NO: 7;
(e) the alpha chain CDR2 of the anti-CCND1 TCR comprises the amino acid sequence as set forth in SEQ ID NO: 6; and
(f) the alpha chain CDR1 of the anti-CCND1 TCR comprises the amino acid sequence as set forth in SEQ ID NO: 5; and
(ii) a second nucleotide sequence, wherein the second nucleotide sequence or the polypeptide encoded by the second nucleotide sequence inhibits the expression of an endogenous TCR.
2 . The nucleic acid molecule of claim 1 , wherein
(i) the alpha chain variable domain of the anti-CCND1 TCR comprises the amino acid sequence of the variable domain present in the amino acid sequence set forth in SEQ ID NO: 1;
(ii) the beta chain variable domain of the anti-CCND1 TCR comprises the amino acid sequence of the variable domain present in the amino acid sequence set forth in SEQ ID NO: 2; or
(iii) both (i) and (ii).
3 . The nucleic acid molecule of claim 1 , wherein:
(a) the anti-CCND1 TCR further comprises an alpha chain constant region, wherein the alpha chain constant region of the anti-CCND1 TCR is different from a constant region of an endogenous alpha chain, and wherein
(i) the alpha chain constant region of the anti-CCND1 TCR comprises an amino acid sequence having at least about 85% sequence identity to a constant region present in the amino acid sequence set forth in SEQ ID NO: 1; or
(ii) the alpha chain constant region of the anti-CCND1 TCR comprises an amino acid sequence comprising at least 1 amino acid substitution relative to a constant region present in the amino acid sequence set forth in SEQ ID NO: 1;
(b) the anti-CCND1 TCR further comprises a beta chain constant region, wherein the beta chain constant region of the anti-CCND1 TCR is different from a constant region of an endogenous beta chain, and wherein
(i) the beta chain constant region of the anti-CCND1 TCR comprises an amino acid sequence having at least about 85% sequence identity to the constant region present in the amino acid sequence set forth in SEQ ID NO: 2; or
(ii) the beta chain constant region comprises an amino acid sequence comprising at least 1, amino acid substitution relative to the constant region present in the amino acid sequence set forth in SEQ ID NO: 2; or
(c) both (a) and (b).
4 . The nucleic acid molecule of claim 1 , wherein the second nucleotide sequence comprises one or more siRNAs that reduce the expression of endogenous TCRs, wherein the one or more siRNAs are complementary to a target sequence within a nucleotide sequence encoding a constant region of the endogenous TCRs.
5 . The nucleic acid molecule of claim 1 , wherein the anti-CCND1 TCR comprises an alpha chain comprising the amino acid sequence set forth in SEQ ID NO: 1, and a beta chain comprising the amino acid sequence set forth in SEQ ID NO: 2.
6 . A cell comprising the nucleic acid molecule of claim 5 .
7 . The cell of claim 6 , which further expresses CD3.
8 . The cell of claim 6 , which is a T cell.
9 . The cell of claim 6 , which is an NK cell, a natural killer T (NKT) cell, or an ILC cell.
10 . A vector comprising the nucleic acid molecule of claim 1 .
11 . A cell comprising the nucleic acid molecule of claim 1 .
12 . The cell of claim 11 , which further expresses CD3.
13 . The cell of claim 11 , which is a T cell.
14 . The cell of claim 11 , which is a natural killer (NK) cell, a natural killer T (NKT) cell, or an ILC cell.
15 . A method of treating a cancer in a subject in need thereof, comprising administering to the subject the cell of claim 11 , wherein the cell is a T cell or an NK cell.
16 . The method of claim 15 , wherein the cancer comprises melanoma, bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease, non-Hodgkin's lymphoma (NHL), primary mediastinal large B cell lymphoma (PMBC), diffuse large B cell lymphoma (DLBCL), follicular lymphoma (FL), transformed follicular lymphoma, splenic marginal zone lymphoma (SMZL), cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, chronic or acute leukemia, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia (ALL) (including non T cell ALL), chronic lymphocytic leukemia (CLL), solid tumors of childhood, lymphocytic lymphoma, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, neoplasm of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma, environmentally induced cancers including those induced by asbestos, other B cell malignancies, or combinations of said cancers.
17 . The method of claim 15 , wherein the cancer is locally advanced, advanced, or metastatic.
18 . The method of claim 15 , wherein the cancer is relapsed or refractory.
19 . The method of claim 15 , wherein the cells are obtained from the subject.