Anti-human papillomavirus 16 E7 T cell receptors
Inventors
Hinrichs, Christian S. • Rosenberg, Steven A.
Assignees
US Department of Health and Human Services
Publication Number
US-10870687-B2
Publication Date
2020-12-22
Expiration Date
2035-05-29
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Abstract
Disclosed is a synthetic T cell receptor (TCR) having antigenic specificity for an HLA-A2-restricted epitope of human papillomavirus (HPV) 16 E7, E711-19. Related polypeptides and proteins, as well as related nucleic acids, recombinant expression vectors, host cells, and populations of cells are also provided. Antibodies, or an antigen binding portion thereof, and pharmaceutical compositions relating to the TCRs of the invention are also provided. Also disclosed are methods of detecting the presence of a condition in a mammal and methods of treating or preventing a condition in a mammal, wherein the condition is cancer, HPV 16 infection, or HPV-positive premalignancy.
Core Innovation
The invention provides synthetic T cell receptors (TCRs) having antigenic specificity for an HLA-A2-restricted epitope of human papillomavirus (HPV) 16 E7, specifically the E711-19 peptide. These TCRs include human variable regions and murine constant regions or functional variants thereof. Related nucleic acids, recombinant expression vectors, host cells, populations of cells, antibodies recognizing the TCRs, and pharmaceutical compositions are also provided. The invention further includes methods for detecting the presence of cancer, HPV 16 infection, or HPV-positive premalignancy in a mammal, as well as methods for treating or preventing these conditions using the inventive TCRs or related materials.
HPV 16 infection is a primary cause of some cancer types such as uterine cervical cancer, and prognosis for HPV-associated cancers is often poor despite advances such as chemotherapy. There is an unmet need for additional and more effective treatments targeting cancer, particularly HPV-associated cancers. The invention addresses this need by providing TCRs that specifically recognize the HPV 16 E7 oncoprotein which is constitutively expressed in cancer cells but not in normal tissues.
The inventive TCRs specifically bind HPV 16 E7 in an MHC class I-dependent manner, preferably with HLA-A2 restriction, enabling immune recognition of HPV 16 E7 expressing tumor cells. The TCRs can destroy multiple types of HPV 16-associated cancer cells while sparing normal cells, potentially reducing toxicity. They show high avidity for the E7 epitope and may successfully treat or prevent cancers which are unresponsive to conventional therapies.
Claims Coverage
The claims cover a synthetic T cell receptor and related compositions and methods, focusing on antigenic specificity for HPV 16 E7 and particular amino acid sequences and structural features. Key inventive features include the combination of human variable and murine constant regions, modifications to improve function, and methods of use in detection and treatment.
TCR comprising a human variable region and murine constant region with HPV 16 E7 specificity
The TCR and its functional variants include the amino acid sequences of SEQ ID NOs: 3-8 and possess antigenic specificity for HPV 16 E7.
HPV 16 E711-19 epitope specificity
The TCR specifically recognizes the HPV 16 E711-19 peptide (SEQ ID NO: 2).
Variable amino acid sequences with leader sequence modification
The TCR comprises amino acid sequences of SEQ ID NOs: 9 and 10, with position 2 of SEQ ID NO: 10 being Ala or Gly.
Substituted constant regions with specific amino acid residues
The TCR includes constant region sequences with substitutions: alpha chain (SEQ ID NO: 16) with variations at positions 48, 112, 114, 115 and beta chain (SEQ ID NO: 18) with variation at position 56.
Combination of constant regions from murine and modified sequences
The TCR comprises any of SEQ ID NOs: 17, 21, 24, 25 for alpha chain and SEQ ID NOs: 19, 23 for beta chain.
Specific combinations of alpha and beta chain sequences including variants
The TCR includes combinations such as SEQ ID NOs: 22, 26, 9 and 24 or 9 and 16 or 9 and 17 for alpha chain, with SEQ ID NOs: 10 and 18 or 20 and 27 for beta chain.
TCR or protein including sequences with linked alpha and beta chains
The TCR or functional variant may comprise the amino acid sequences of SEQ ID NO: 29 or 30 linked by a peptide linker or fusion protein.
Isolated polypeptides comprising complementarity determining regions
Polypeptides comprising amino acid sequences of SEQ ID NOs: 3-5, 6-8, or 3-8 which represent CDRs of the TCR.
Isolated polypeptides comprising variable regions
Polypeptides comprising amino acid sequences of SEQ ID NO: 9, SEQ ID NO: 10 (position 2 Ala or Gly), or both, isolated or purified.
Polypeptides comprising combined variable and constant regions
Polypeptides comprising combinations of defined sequences including SEQ ID NOs: 9, 10, 16, 17, 20, 22, 24, 26, 27, 29, and 30 as isolated or purified proteins.
Proteins comprising alpha and beta polypeptide chains with defined sequences
Proteins with first polypeptide chain comprising SEQ ID NOs: 3-5 and second polypeptide chain comprising SEQ ID NOs: 6-8.
Proteins with variable region sequences in alpha and beta chains
First chain includes SEQ ID NO: 9; second includes SEQ ID NO: 10 with position 2 Ala or Gly; representing isolated or purified protein.
Proteins combining specific variable and constant region sequences
Proteins combining sequences such as SEQ ID NO: 22 or 26 with beta chain sequences SEQ ID NOs: 10 and 18 or 20 and 27.
Fusion proteins or recombinant antibodies including the defined TCR sequences
Proteins comprising SEQ ID NO: 29 or 30 and configured as fusion proteins or recombinant antibodies.
Pharmaceutical compositions comprising the inventive TCR or functional variant
Pharmaceutical compositions containing the inventive TCR or functional variant with a pharmaceutically acceptable carrier.
Method of detecting cancer, HPV 16 infection or HPV-positive premalignancy
A method involving contacting a sample with the inventive TCR or functional variant and detecting complex formation indicative of the condition.
Method of treating or preventing HPV-associated cancer
Administering the inventive TCR or functional variant in an effective amount to treat or prevent cancer, HPV 16 infection, or HPV-positive premalignancy.
Method specifying cancer sites
Wherein the treated or prevented cancer is of uterine cervix, oropharynx, anus, anal canal, anorectum, vagina, vulva, or penis.
Pharmaceutical compositions with specific TCR sequences
Compositions comprising specific TCR sequences of claim 2, 3, 8, 9, or 11 with carriers.
The claims comprehensively cover TCRs having human variable and murine constant regions with specificity for HPV 16 E7, particularly the E711-19 epitope, as well as variants with amino acid substitutions that improve function. Related polypeptides, proteins, fusion proteins, pharmaceutical compositions, and methods for detection and treatment of HPV-associated conditions are also claimed.
Stated Advantages
Ability to destroy cells of multiple types of HPV 16-associated cancer by targeting HPV 16 E7 expressed in various cancer types.
Selective targeting of cancer cells expressing HPV 16 E7 while sparing normal cells, thereby reducing toxicity.
Potential to successfully treat or prevent HPV-positive cancers that do not respond to conventional treatments such as chemotherapy, surgery, or radiation.
High avidity recognition of HPV 16 E7 enabling detection and immune activation against unmanipulated tumor cells without additional treatment or modification.
Documented Applications
Detecting the presence of cancer, HPV 16 infection, or HPV-positive premalignancy in a mammal by forming and detecting a complex with the inventive TCR or related materials.
Treating or preventing cancer, HPV 16 infection, or HPV-positive premalignancy in mammals by administering the inventive TCR, nucleic acids encoding it, recombinant vectors, or host cells expressing the TCR.
Treatment of various HPV 16-positive cancers, including cancers of the uterine cervix, oropharynx, anus, anal canal, anorectum, vagina, vulva, and penis.
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