T cell receptors recognizing HLA-Cw8 restricted mutated KRAS
Inventors
Tran, Eric • Lu, Yong-Chen • Rosenberg, Steven A.
Assignees
US Department of Health and Human Services
Publication Number
US-10556940-B2
Publication Date
2020-02-11
Expiration Date
2036-09-09
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Abstract
Disclosed is an isolated or purified T cell receptor (TCR) having antigenic specificity for mutated Kirsten rat sarcoma viral oncogene homolog (KRAS) presented in the context of an HLA-Cw*0802 molecule. Related polypeptides and proteins, as well as related nucleic acids, recombinant expression vectors, host cells, populations of cells, and pharmaceutical compositions are also provided. Also disclosed are methods of detecting the presence of cancer in a mammal and methods of treating or preventing cancer in a mammal.
Core Innovation
The invention provides an isolated or purified T cell receptor (TCR) that has antigenic specificity for mutated Kirsten rat sarcoma viral oncogene homolog (KRAS) presented in the context of a human leukocyte antigen (HLA)-Cw8 molecule, preferably HLA-Cw*0802. The invention includes related polypeptides, proteins, nucleic acids, recombinant expression vectors, host cells, cell populations, and pharmaceutical compositions. Methods for detecting cancer presence and methods for treating or preventing cancer in a mammal using these TCRs are also provided.
Mutated KRAS, especially the G12D mutation in KRAS variants A or B, is implicated in multiple cancers such as pancreatic, colorectal, lung adenocarcinoma, endometrial, epithelial ovarian, and prostate cancers. The inventive TCRs specifically recognize mutated KRAS peptides of various lengths, including the crucial epitope GADGVGKSA (SEQ ID NO: 18), in an HLA-Cw8-dependent manner. The TCRs bind to mutated KRAS presented by HLA-Cw8 molecules encoded by multiple alleles, expanding applicability.
The background highlights the poor prognosis and limited treatments for various cancers when they become metastatic and unresectable. This underscores the unmet need for additional cancer treatments. The invention addresses this need by providing TCRs that target mutated KRAS specifically, aiming to selectively destroy cancer cells expressing mutated KRAS with minimized off-target toxicity to normal cells. This specificity is achieved through HLA-Cw*0802 restriction and targeting the mutation-associated epitope, enabling immunotherapy for patients expressing this allele and harboring mutated KRAS-positive tumors.
Claims Coverage
The patent includes multiple independent claims focusing on isolated or purified TCRs, polypeptides, proteins, nucleic acids, recombinant vectors, host cells, methods of detection, and treatment related to mutated KRAS presented by HLA-Cw8 molecules.
TCR complementarity determining regions (CDRs) specific for mutated KRAS
An isolated or purified TCR comprising α chain CDR1 (SEQ ID NO: 3), CDR2 (SEQ ID NO: 4), and CDR3 (SEQ ID NO: 5), and β chain CDR1 (SEQ ID NO: 6), CDR2 (SEQ ID NO: 7), and CDR3 (SEQ ID NO: 8).
TCR variable regions comprising specified sequences
TCRs comprising α chain variable region (SEQ ID NO: 9), β chain variable region (SEQ ID NO: 10), or both, conferring antigenic specificity for mutated KRAS.
TCR comprising substituted constant regions with specific amino acid substitutions
TCRs with α chain constant region (SEQ ID NO: 11) exhibiting substitutions at positions 48 (Thr or Cys), 112, 114, and 115 with specified amino acids, and β chain constant region (SEQ ID NO: 12) with substitution at position 57 (Ser or Cys), alone or combined.
TCR comprising full-length α and β chains with substitutions
TCRs comprising α chain (SEQ ID NO: 13) and β chain (SEQ ID NO: 14) with specified amino acid substitutions at defined positions conferring antigenic specificity.
Polypeptides and proteins comprising functional portions of the inventive TCRs
Isolated or purified polypeptides and proteins comprising the above CDRs or variable regions, optionally including substituted constant regions, retaining the ability to specifically bind mutated KRAS presented with HLA-Cw8.
Nucleic acids and recombinant vectors encoding the inventive TCRs
Nucleotide sequences encoding any of the claimed TCRs, polypeptides, or proteins, incorporated into recombinant expression vectors suitable for host cell expression.
Host cells and populations expressing the inventive TCRs
Isolated or purified host cells comprising the recombinant vectors encoding the TCRs, enabling expression and use in therapeutic or diagnostic contexts.
Pharmaceutical compositions comprising the inventive TCRs
Pharmaceutical compositions containing the claimed TCRs together with pharmaceutically acceptable carriers for administration.
Methods of detecting cancer expressing HLA-Cw8 and KRAS G12D
Methods involving contacting a sample comprising cancer cells with the claimed TCRs to form detectable complexes indicative of cancer presence.
Methods of treating or preventing HLA-Cw8 and KRAS G12D expressing cancers
Administering the claimed TCRs or related compositions in effective amounts to treat or prevent such cancers, including pancreatic, colorectal, lung, endometrial, ovarian, or prostate cancer.
The independent claims cover TCRs and their functional portions defined by precise CDR, variable, and constant region sequences specific for mutated KRAS presented by HLA-Cw8, nucleic acids encoding these TCRs, recombinant vectors, host cells expressing them, pharmaceutical compositions for therapy, and methods for detection and treatment of relevant cancers.
Stated Advantages
The inventive TCRs selectively target mutated KRAS expressed by cancer cells and not normal cells, minimizing or eliminating toxicity.
They provide potential to treat or prevent mutated KRAS-positive cancers unresponsive to conventional treatments like chemotherapy, surgery, or radiation.
The TCRs exhibit high avidity, enabling recognition of unmanipulated tumor cells without prior treatment or manipulation.
Expression of the HLA-Cw*0802 allele in significant ethnic populations (up to about 11%) broadens the scope of patients eligible for immunotherapy using these TCRs.
Documented Applications
Detecting the presence of cancer in mammals by contacting samples with the inventive TCRs to form detectable complexes.
Treating or preventing cancer in mammals, specifically cancers expressing mutated KRAS G12D in the context of HLA-Cw8.
Therapeutic targeting of cancers including pancreatic carcinoma, colorectal cancer, lung adenocarcinoma, endometrial cancer, epithelial ovarian cancer, and prostate cancer using T cells genetically engineered to express the inventive TCRs.
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