T-cell modulatory multimeric polypeptides and methods of use thereof
The present disclosure provides variant immunomodulatory polypeptides, and fusion polypeptides comprising the variant immunomodulatory peptides. The present disclosure provides T-cell modulatory multimeric polypeptides, and compositions comprising same, where the T-cell modulatory multimeric polypeptides comprise a variant immunomodulatory polypeptide of the present disclosure. The present disclosure provides nucleic acids comprising nucleotide sequences encoding the T-cell modulatory multimeric polypeptides, and host cells comprising the nucleic acids. The present disclosure provides methods of modulating the activity of a T cell; the methods comprise contacting the T cell with a T-cell modulatory multimeric polypeptide of the present disclosure.
1. A multimeric polypeptide comprising a heterodimer comprising:
a) a first polypeptide comprising:
i) a human papillomavirus epitope; and
ii) first class I major histocompatibility complex (MHC) polypeptide, wherein the first MHC polypeptide is a β2-microglobulin (β2M) polypeptide;
b) a second polypeptide comprising:
i) a second class I MHC polypeptide, wherein the second MHC polypeptide is an MHC Class I HLA-A heavy chain polypeptide;
ii) at least one immunomodulatory polypeptide, wherein the at least one immunomodulatory polypeptide is an IL-2 variant polypeptide that comprises an amino acid sequence having at least 98% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:44, wherein amino acid 16 is Ala and amino acid 42 is Ala, wherein the percent sequence identity is determinable by a sequence alignment performed using BLAST; and
iii) an immunoglobulin (Ig) Fc polypeptide.
2. A multimeric polypeptide of claim 1 , wherein the HPV epitope is an HPV E7 epitope.
3. A multimeric polypeptide of claim 1 , wherein the β2M polypeptide comprises an amino acid sequence having at least 95% percent amino acid sequence identity to amino acids 21 to 119 of SEQ ID NO:95.
4. A multimeric polypeptide of claim 1 , wherein the β2M polypeptide comprises an amino acid sequence as set forth in SEQ ID NO:17.
5. A multimeric polypeptide of claim 1 wherein the HLA-A heavy chain polypeptide comprises an amino acid sequence having at least 95% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:19, wherein the percent sequence identity is determinable by a sequence alignment performed using BLAST.
6. A multimeric polypeptide of claim 1 , wherein the HLA-A heavy chain polypeptide comprises a Cys at residue 236 of the MHC class I HLA-A heavy chain polypeptide based on the numbering depicted in SEQ ID NO:19.
7. A multimeric polypeptide of claim 1 , wherein the Ig Fc polypeptide comprises an amino acid sequence having at least about 95% percent amino acid sequence identity to SEQ ID NO:57, wherein the percent sequence identity is determinable by a sequence alignment performed using BLAST.
8. A multimeric polypeptide of claim 7 , wherein the Ig Fc polypeptide comprises an L14A substitution and an L15A substitution based on the amino acid numbering depicted in SEQ ID NO:57.
9. A multimeric polypeptide of claim 1 , wherein the multimeric polypeptide comprises two copies of the IL-2 variant polypeptide, and wherein each copy comprises the amino acid sequence of SEQ ID NO:84.
10. A multimeric polypeptide of claim 9 , wherein the first polypeptide comprises a peptide linker between the epitope and the β2M polypeptide, and
wherein the second polypeptide comprises an independently selected peptide linker between one or more of:
a) a first copy of the IL-2 variant polypeptide and a second copy of the IL-2 variant polypeptide;
b) one of the two copies of the IL-2 polypeptide and the MHC class I heavy chain polypeptide; and
c) the MHC class I heavy chain polypeptide and the Ig Fc polypeptide.
11. A multimeric polypeptide of claim 1 , wherein the first and second polypeptides are covalently linked to one another via a disulfide bond between a Cys residue at amino acid 12 of the β2M polypeptide and a Cys residue at amino acid 236 of the class I MHC heavy chain polypeptide.
12. A multimeric polypeptide of claim 9 , wherein:
a) the first polypeptide comprises, in order from N-terminus to C-terminus:
i) the HPV epitope;
ii) a peptide linker; and
iii) the β2M polypeptide, wherein the β2M polypeptide comprises an amino acid sequence having at least 95% percent amino acid sequence identity to amino acids 21 to 119 of SEQ ID NO:95;
b) the second polypeptide comprises, in order from N-terminus to C-terminus:
i) the IL-2 variant polypeptide;
ii) a peptide linker;
iii) the IL-2 variant polypeptide;
iv) an optional peptide linker;
v) the HLA-A heavy chain polypeptide, wherein the HLA-A heavy chain polypeptide comprises an amino acid sequence having at least 95% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:19;
vi) an optional peptide linker; and
vii) the Ig Fc polypeptide, wherein the Ig Fc polypeptide is an IgG1 Fc polypeptide that comprises an amino acid sequence having at least about 95% percent amino acid sequence identity to SEQ ID NO:57,
wherein the peptide linkers are independently selected, and wherein the percent sequence identity is determinable by a sequence alignment performed using BLAST.
13. A multimeric polypeptide of claim 12 wherein the first and second polypeptides are covalently linked to one another via a disulfide bond between a Cys residue at amino acid 12 of the β2M polypeptide and a Cys residue at amino acid 236 of the HLA-A polypeptide.
14. A multimeric polypeptide of claim 12 , wherein the HPV epitope is an HPV E7 epitope.
15. A homodimer comprising two heterodimers according to claim 1 .
16. A homodimer comprising two heterodimers according to claim 12 .
17. A homodimer comprising two heterodimers according to claim 13 .
18. A homodimer comprising two heterodimers according to claim 14 .
19. A pharmaceutical composition comprising a homodimer of claim 15 .
20. A pharmaceutical composition comprising a homodimer of claim 16 .
21. A pharmaceutical composition comprising a homodimer of claim 17 .
22. A pharmaceutical composition comprising a homodimer of claim 18 .
23. A method of treating an HPV-associated cancer or an HPV-attributable cancer in a patient in need of such treatment, the method comprising administering a pharmaceutical composition of claim 20 to the patient in an amount effective to treat said cancer.
24. A method of treating an HPV-associated cancer or an HPV-attributable cancer in a patient in need of such treatment, the method comprising administering a pharmaceutical composition of claim 20 to the patient in an amount effective to treat said cancer.
25. A method of treating an HPV-associated cancer or an HPV-attributable cancer in a patient in need of such treatment, the method comprising administering a pharmaceutical composition of claim 21 to the patient in an amount effective to treat said cancer.
26. A method of treating an HPV-associated cancer or an HPV-attributable cancer in a patient in need of such treatment, the method comprising administering a pharmaceutical composition of claim 22 to the patient in an amount effective to treat said cancer.