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 heterodimer comprising:
a) a first polypeptide comprising:
i) a cancer-associated epitope other than a human papillomavirus epitope; and
ii) a first major histocompatibility complex (MHC) Class I polypeptide, wherein the first MHC polypeptide is a β2-microglobulin (β2M) polypeptide comprising an amino acid sequence having at least 95% percent amino acid sequence identity to amino acids 21 to 119 of SEQ ID NO:95, wherein the percent sequence identity is determinable by a sequence alignment performed using BLAST, and
b) a second polypeptide comprising:
i) two copies of an IL-2 polypeptide, each copy comprising the amino acid sequence of SEQ ID NO: 84, wherein the two IL-2 polypeptides are linked by a peptide linker;
ii) a second MHC polypeptide, wherein the second MHC polypeptide is an MHC Class I HLA-A heavy chain polypeptide; and
iii) an immunoglobulin (Ig) Fc polypeptide, and
wherein the heterodimer comprises a disulfide bond that covalently links a Cys residue in the β2M polypeptide to a Cys residue in the MHC class I heavy chain polypeptide.
2. A heterodimer according to claim 1 , wherein
the Ig Fc polypeptide is an IgG1 Fc polypeptide 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, and comprises an L14A substitution and an LISA substitution based on the amino acid numbering depicted in SEQ ID NO:57,
the second polypeptide comprises a peptide linker between one or more of:
a) a first copy of the IL-2 polypeptide and a second copy of the IL-2 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 IgG1 Fc polypeptide.
3. A heterodimer according to claim 1 , wherein
the first and second polypeptides are covalently linked to one another via a disulfide bond between (i) a Cys residue at amino acid 12 of the β2M) polypeptide and (ii) 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.
4. A homodimer comprising two heterodimers according to claim 1 , wherein the two heterodimers are joined to each other by one or more disulfide bonds that join the Ig Fc polypeptide of one heterodimer to the Ig Fc polypeptide of the other heterodimer.
5. A homodimer comprising two heterodimers according to claim 2 , wherein the two heterodimers are joined to each other by one or more disulfide bonds that join the Ig Fc polypeptide of one heterodimer to the Ig Fc polypeptide of the other heterodimer.
6. A homodimer comprising two heterodimers according to claim 3 , wherein the two heterodimers are joined to each other by one or more disulfide bonds that join the Ig Fc polypeptide of one heterodimer to the Ig Fc polypeptide of the other heterodimer.
7. A pharmaceutical composition comprising a heterodimer of claim 1 .
8. A pharmaceutical composition comprising a heterodimer of claim 2 .
9. A pharmaceutical composition comprising a heterodimer of claim 3 .
10. A pharmaceutical composition comprising a homodimer of claim 4 .
11. A pharmaceutical composition comprising a homodimer of claim 5 .
12. A pharmaceutical composition comprising a homodimer of claim 6 .
13. A method of treating cancer in a patient in need of such treatment, the method comprising administering a pharmaceutical composition of claim 10 to the patient in an amount effective to treat said cancer.
14. The method according to claim 13 , wherein the homodimer is administered in an amount of from about 1 mg/kg to about 5 mg/kg of body weight.
15. The method according to claim 13 , wherein the homodimer is administered in an amount of from about 5 mg/kg to about 10 mg/kg of body weight.
16. A method of treating cancer in a patient in need of such treatment, the method comprising administering a pharmaceutical composition of claim 11 to the patient in an amount effective to treat said cancer.
17. The method according to claim 16 , wherein the homodimer is administered in an amount of from about 1 mg/kg to about 5 mg/kg of body weight.
18. The method according to claim 16 , wherein the homodimer is administered in an amount of from about 5 mg/kg to about 10 mg/kg of body weight.
19. A method of treating cancer in a patient in need of such treatment, the method comprising administering a pharmaceutical composition of claim 12 to the patient in an amount effective to treat said cancer.
20. The method according to claim 19 , wherein the homodimer is administered in an amount of from about 1 mg/kg to about 5 mg/kg of body weight.
21. The method according to claim 19 , wherein the homodimer is administered in an amount of from about 5 mg/kg to about 10 mg/kg of body weight.