Brachyury polypeptides and methods for use
Inventors
Schlom, Jeffrey • Palena, Claudia M. • Kozlov, Andrei P. • Tsang, Kwong-Yok
Assignees
US Department of Health and Human Services
Publication Number
US-8188214-B2
Publication Date
2012-05-29
Expiration Date
2028-02-27
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Abstract
It is disclosed herein that Brachyury is expressed in human tumors, specifically in tumors of the small intestine, stomach, kidney, bladder, uterus, ovary, and testes, as well as in lung, colon and prostate carcinomas. Immunogenic Brachyury polypeptides are disclosed herein. These polypeptides can be used in diagnostic assays for Brachyury expression, as well as for inducing an immune response to Brachyury. Polynucleotides encoding the immunogenic Brachyury polypeptides, vectors including these polypeptides, host cells transformed with these vectors, and methods of using these polypeptides, polynucleotides, vectors, and host cells are provided. Methods of diagnosing a Brachyury-expressing cancer are also provided. Exemplary cancers include lung, colon, small intestine, stomach, kidney, bladder, uterus, ovary, and testes and prostate cancers. Methods of treating cancer are also disclosed.
Core Innovation
Brachyury is expressed in human tumors including those of the small intestine, stomach, kidney, bladder, uterus, ovary, testes, lung, colon, and prostate carcinomas, as well as in chronic lymphocytic leukemia and other B cell malignancies. Immunogenic Brachyury polypeptides that can induce an immune response to Brachyury are disclosed. These polypeptides can be used in diagnostic assays for Brachyury expression and in therapeutic methods, including compositions, vectors, and host cells expressing these Brachyury polypeptides.
The invention provides methods for diagnosing Brachyury-expressing cancers using these immunogenic polypeptides, nucleic acids encoding the polypeptides, or antibodies specific to the polypeptides. Methods for inducing an immune response to tumors expressing Brachyury include administering immunogenic peptides, nucleic acids, or viral vectors encoding these polypeptides, optionally in conjunction with co-stimulatory molecules, cytokines, or traditional cancer therapies such as surgery, radiation, or chemotherapy.
The background identifies the problem of the need for additional tumor antigens to aid in the diagnosis and immunotherapy of various cancers. Existing immunotherapies based on known tumor antigens (e.g., PSA, PSMA) are limited, and more antigens are required to improve cancer detection and treatment. Brachyury, implicated in embryonic development, was identified through computer-based EST analysis as specifically expressed in tumor tissues but not in normal tissues, indicating its potential as a novel cancer antigen target for diagnosis and immunotherapy.
Claims Coverage
The claims include multiple independent claims directed to isolated Brachyury polypeptides having specific amino acid sequences, nucleic acids encoding these polypeptides, vectors, and compositions including recombinant viruses. The main inventive features focus on the specific immunogenic Brachyury peptide sequences and their use.
Isolated immunogenic Brachyury polypeptides not exceeding twelve amino acids
Isolated polypeptides comprising specific amino acid sequences of Brachyury, including sequences such as WLLPGTSTX1 (SEQ ID NO: 3), SX2YX3SLX4SX5 (SEQ ID NO: 18), WLLX6GTSTX7 (SEQ ID NO: 19), X8LIASWTPV (SEQ ID NO: 20), X9LIASX10TPV (SEQ ID NO: 21), and ALYSFLLDFV (SEQ ID NO: 22), where variable residues are leucine or valine, proline, serine, threonine, leucine, valine, tryptophan, isoleucine, tyrosine, arginine, lysine, and others as specified, and where the polypeptides are no more than twelve amino acids in length.
Polynucleotides encoding the isolated Brachyury polypeptides
Nucleic acid sequences encoding the immunogenic Brachyury polypeptides (not encoding more than twelve amino acids of the full-length SEQ ID NO: 1), optionally operably linked to promoters for expression.
Vectors and host cells comprising nucleic acids encoding the immunogenic Brachyury polypeptides
Expression vectors, including plasmid and viral vectors (e.g., retrovirus, orthopox, avipox, fowlpox, capripox, suipox, adenoviral, herpes virus, alpha virus, baculovirus, Sindbis virus, vaccinia virus, and poliovirus vectors), and host cells transformed with these vectors for expression of the Brachyury polypeptides.
Compositions comprising recombinant viruses coexpressing Brachyury polypeptides and costimulatory molecules
Compositions with a first recombinant virus encoding the Brachyury polypeptide and a second recombinant virus encoding one or more costimulatory molecules such as B7-1, B7-2, LFA-3, or ICAM-1, that coinfect host cells resulting in coexpression of the polypeptide and costimulatory molecules.
Methods for eliciting an immune response using the immunogenic polypeptides or encoding nucleic acids
Methods of eliciting an immune response in a subject by administering a therapeutically effective amount of the isolated Brachyury polypeptides or nucleic acids encoding them, optionally with adjuvants or immunostimulatory molecules like IL-2, GM-CSF, TNF-α, and others.
Methods of detecting CD8+ T cells specific for Brachyury
Use of reagents comprising the immunogenic Brachyury polypeptides to detect CD8+ T cells that specifically recognize Brachyury (SEQ ID NO: 1) in a subject.
The independent claims cover isolated immunogenic Brachyury polypeptides of specific sequences up to twelve amino acids in length, nucleic acids encoding these polypeptides, vectors and host cells for expression, compositions comprising recombinant viruses encoding these polypeptides along with costimulatory molecules, and methods for eliciting immune responses and detecting specific T cells to Brachyury in subjects.
Stated Advantages
Brachyury polypeptides provide novel tumor-associated antigens expressed in a variety of human tumors but not in most normal tissues, enabling specific targeting.
The use of immunogenic Brachyury peptides enables induction of cytotoxic T lymphocyte responses against tumor cells expressing Brachyury.
Compositions including recombinant viruses coexpressing Brachyury and costimulatory molecules enhance immunogenicity and tumor rejection potential.
Methods for diagnosing Brachyury-expressing cancers improve specificity and can aid in identifying tumor origin and progression.
Documented Applications
Use of immunogenic Brachyury polypeptides for inducing immune responses in subjects with tumors expressing Brachyury, including tumors of the small intestine, stomach, kidney, bladder, uterus, ovary, testes, lung, colon, and prostate.
Diagnostic methods for detecting Brachyury expression in tumors using immunogenic peptides, nucleic acids, or antibodies.
Therapeutic methods of treating cancer by administering immunogenic Brachyury polypeptides, nucleic acids, or viral vectors encoding these polypeptides.
Use of inhibitory nucleic acids (e.g., siRNA, antisense) to reduce Brachyury expression for treatment of tumors, including B cell malignancies such as chronic lymphocytic leukemia, Burkitt's lymphoma, and Hodgkin's lymphoma.
Methods involving dendritic cells pulsed with Brachyury polypeptides for immunotherapy.
Pharmaceutical compositions comprising the disclosed polypeptides, nucleic acids, or host cells for cancer immunotherapy.
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