Meditope-enabled T cells
Provided herein are compositions which exhibit novel therapeutic capabilities and allow to reduce the off-target effect of therapeutic antibodies. The compositions include recombinant proteins that if expressed by a T cell can efficiently recruit therapeutic antibodies to their site of action.
1 . A recombinant protein comprising:
(i) a meditope capable of binding a meditope-enabled Fab;
(ii) an intracellular T-cell signaling domain; and
(iii) a transmembrane domain connecting said meditope to said intracellular T-cell signaling domain; wherein said recombinant protein does not comprise an antigen binding domain.
2 . The recombinant protein of claim 1 , further comprising a spacer region connecting said meditope to said transmembrane domain.
3 . The recombinant protein of claim 2 , wherein said spacer region is a constant heavy chain 3 (CH3) domain.
4 . The recombinant protein of claim 3 , further comprising a peptide linker connecting said meditope to said spacer region.
5 . The recombinant protein of claim 1 , further comprising a detectable domain bound to the C-terminus of said intracellular T-cell signaling domain.
6 . The recombinant protein of claim 5 , further comprising a self-cleaving peptidyl sequence connecting said intracellular T-cell signaling domain to said detectable domain.
7 . An isolated nucleic acid encoding a recombinant protein of claim 1 .
8 . An expression vector comprising the nucleic acid of claim 7 .
9 . A T lymphocyte comprising the expression vector of claim 8 .
10 . A T lymphocyte comprising the recombinant protein of claim 1 .
11 . A method of treating cancer, said method comprising administering to a subject in need thereof an effective amount of the T lymphocyte of claim 9 and an antigen-binding domain capable of binding to said meditope, wherein said antigen-binding domain is a cetuximab meditope-enabled domain, trastuzumab meditope-enabled domain, pertuzumab meditope-enabled domain, M5A meditope-enabled domain or rituximab meditope-enabled domain.
12 . The method of claim 11 , wherein said T-lymphocyte and said antigen-binding domain are administered simultaneously or sequentially.
13 . The method of claim 11 , said method comprising:
(i) prior to said administering allowing said meditope to bind said antigen-binding domain in vitro, thereby forming a T-lymphocyte-recombinant protein complex; and
(ii) administering said T-lymphocyte-recombinant protein complex to said subject, thereby treating cancer in said subject.
14 . The recombinant protein of claim 2 , wherein said recombinant protein is a first recombinant protein, said meditope is a first meditope, said intracellular T-cell signaling domain is a first intracellular T-cell signaling domain, said transmembrane domain is a first transmembrane domain and said spacer region is a first spacer region.
15 . The recombinant protein of claim 14 , wherein said first recombinant protein is non-covalently bound to a second recombinant protein, said second recombinant protein comprising:
(i) a second meditope;
(ii) a second intracellular T-cell signaling domain;
(iii) a second transmembrane domain connecting said meditope to said second intracellular T-cell signaling domain; and
(iv) a second spacer region, wherein said second spacer region connects said second meditope to said second transmembrane domain, wherein said first spacer region is non-covalently bound to said second spacer region; wherein said second recombinant protein does not comprise an antigen binding domain.
16 . The recombinant protein of claim 15 , wherein said first meditope and said second meditope are chemically different or chemically the same.
17 . The recombinant protein of claim 1 , wherein said meditope has the formula:
X0-X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12 (I),
wherein
X0 is Ser or null;
X1 is Ser, Cys, Gly, β-alanine, diaminopropionic acid, β-azidoalanine, or null;
X2 is Gln or null, X3 is Phe, Tyr, β,β′-diphenyl-Ala, His, Asp, 2-bromo-L-phenylalanine, 3-bromo-L-phenylalanine, 4-bromo-L-phenylalanine, Asn, Gln, a modified Phe, a hydratable carbonyl-containing residue, or a boronic acid-containing residue;
X4 is Asp or Asn;
X5 is Leu, β,β′-diphenyl-Ala, Phe, Trp, Tyr, a non-natural analog of phenylalanine, tryptophan, or tyrosine, a hydratable carbonyl-containing residue, or a boronic acid-containing residue;
X6 is Cys or Ser;
X7 is Cys, Thr, or Ser;
X8 is protected Arg, Arg, or Ala;
X9 is Cys, Arg or Ala;
X10 is Leu, Gln, Glu, β,β′-diphenyl-Ala, Phe, Trp, Tyr, a non-natural analog of phenylalanine, tryptophan, or tyrosine, a hydratable carbonyl-containing residue, or a boronic acid-containing residue;
X11 is Cys, Gln, Lys or Arg;
X12 is Ser, Cys, Gly, 7-aminoheptanoic acid, β-alanine, diaminopropionic acid, propargylglycine, isoaspartic acid, or null; and
X1 and X12 are optionally joined together to form a cyclic peptidyl moiety.
18 . The recombinant protein of claim 17 , wherein the VL region of said meditope-enabled Fab comprises a threonine at position 40, an asparagine at position 41, and an aspartate at position 85, according to Kabat numbering, and the VH region of said meditope-enabled Fab comprises a serine or proline at position 40 and an isoleucine, tyrosine, methionine, phenylalanine, or a tryptophan at position 89, according to Kabat numbering.