HLA class I-restricted t cell receptors against mutated RAS
Inventors
Yoseph, Rami • Lu, Yong-Chen • Cafri, Gal • Rosenberg, Steven A.
Assignees
US Department of Health and Human Services
Publication Number
US-11466071-B2
Publication Date
2022-10-11
Expiration Date
2038-12-03
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Abstract
Disclosed is an isolated or purified T cell receptor (TCR), wherein the TCR has antigenic specificity for a mutated RAS amino acid sequence presented by a human leukocyte antigen (HLA) Class I 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 isolated or purified T cell receptors (TCRs) that have antigenic specificity for mutated human RAS amino acid sequences when presented by a human leukocyte antigen (HLA) Class I molecule. These mutated sequences include mutated forms of human Kirsten rat sarcoma viral oncogene homolog (KRAS), Harvey rat sarcoma viral oncogene homolog (HRAS), or Neuroblastoma rat sarcoma viral oncogene homolog (NRAS). The TCRs can comprise amino acid sequences corresponding to SEQ ID NOs: 1-6 and have the capacity to recognize mutated RAS peptides, particularly those with a substitution of glycine at position 12, such as G12V mutation.
The problem addressed by the invention originates from the limited treatment options available for cancers, especially when they become metastatic and unresectable. Despite advances in surgery, chemotherapy, and radiation therapy, prognoses for cancers including pancreatic, colorectal, lung, endometrial, ovarian, and prostate cancers remain poor. There exists an unmet need for additional treatments that specifically target cancer cells expressing mutated RAS while sparing normal cells.
The invention further discloses related polypeptides, proteins, nucleic acids, recombinant expression vectors, host cells, and pharmaceutical compositions that utilize the inventive TCR. Methods for detecting cancer presence and for treating or preventing cancer in mammals using these TCRs or related biological materials are also provided. Notably, the invention includes TCRs engineered with modifications in constant regions to enhance expression, reduce mispairing, and improve anti-tumor activity, and TCRs comprising either human or murine constant regions or chimeric combinations thereof.
Claims Coverage
The patent includes one independent claim directed to an isolated or purified nucleic acid encoding a TCR specific for mutated human RAS presented by HLA Class I molecules, and further claims cover recombinant vectors, host cells, populations of cells, pharmaceutical compositions, and methods of detecting and treating cancer using the TCRs.
Isolated nucleic acid encoding a TCR with specific amino acid sequences
An isolated or purified nucleic acid comprising a nucleotide sequence encoding a T cell receptor (TCR) that includes the amino acid sequences of SEQ ID NOs: 1-3, 4-6, or 1-6, which confer antigenic specificity for mutated human RAS amino acid sequences.
TCR specificity for mutated RAS peptides presented by HLA Class I molecules
The TCR encoded by the nucleic acid has antigenic specificity for mutated human RAS amino acid sequences (mutated KRAS, HRAS, or NRAS), especially peptides with a G12V mutation, presented by human leukocyte antigen (HLA) Class I molecules, including HLA-A, HLA-A11, and preferably HLA-A*11:01.
Inclusion of variable and constant regions with specific substitutions
The nucleic acid encodes TCRs comprising human variable regions (SEQ ID NOs: 7 and 8) and murine or substituted murine constant regions (SEQ ID NOs: 17, 18, 21, and 22) with defined amino acid substitutions including cysteine and hydrophobic residues to enhance TCR properties such as expression, stability, and reduced mispairing.
Recombinant expression vectors and host cells
Recombinant expression vectors comprising the nucleic acid encoding the TCR are claimed, as well as isolated or purified host cells and populations of cells transformed or transduced with such vectors capable of expressing the inventive TCR.
Pharmaceutical compositions and methods for cancer detection and treatment
Pharmaceutical compositions comprising the host cells and methods of detecting the presence of cancer in a mammal by contacting cancer cells with the host cells expressing the TCR and detecting complex formation are claimed. Methods of treating cancer in mammals by administering the host cells are also claimed.
The claims cover nucleic acids encoding TCRs specific for mutated RAS peptides presented by HLA Class I molecules, recombinant vectors, host cells expressing these TCRs, pharmaceutical compositions, and methods of detecting and treating cancer, including embodiments with modified constant regions to improve functionality.
Stated Advantages
The inventive TCRs target mutated RAS expressed by cancer cells but not by normal cells, potentially reducing toxicity.
The TCRs may successfully treat or prevent mutated RAS-positive cancers unresponsive to chemotherapy, surgery, or radiation.
High avidity recognition of mutated RAS enables recognition of unmanipulated tumor cells.
The use of HLA-A*11:01 targeting TCRs increases immunotherapy eligibility among diverse ethnic groups.
Human amino acid sequences in the TCRs may reduce risk of immune rejection compared to mouse sequences.
Documented Applications
Methods of detecting the presence of cancer in a mammal by contacting samples with the inventive TCRs or cells expressing them, detecting complex formation indicative of cancer presence.
Methods of treating or preventing cancer in a mammal by administering pharmaceutical compositions, nucleic acids, recombinant vectors, or host cells expressing the inventive TCRs.
Use in adoptive cell transfer therapies utilizing host cells expressing the TCRs to target mutated RAS-positive cancer cells.
Treatment and detection of cancers expressing mutated KRAS, HRAS, or NRAS, including pancreatic, colorectal, lung (adenocarcinoma), endometrial, ovarian (epithelial ovarian cancer), and prostate cancers.
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