Fusion constructs and methods of using thereof
Provided herein is a composition comprising a fusion protein or a fragment or a variant thereof comprising an anti-PD1 antibody or a fragment/variant thereof and a TGF-β trap. Provided herein is a composition comprising a fusion protein or a fragment thereof or a variant thereof comprising an anti-PD1 antibody or a fragment/variant thereof and a ADA2 polypeptide. Also provided herein are methods of using the composition in treating cancer.
1 . A method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a fusion protein comprising:
(a) an antibody, or an antigen-binding fragment or variant thereof, that specifically binds to programmed cell death protein-1 (PD-1) and comprises either:
(1) a VH region having the sequence of SEQ ID NO: 6 and a VL region having the sequence of SEQ ID NO: 12; or
(ii) a VH region having the sequence of SEQ ID NO: 7 and a VL region having the sequence of SEQ ID NO: 13;
and
(b) a transforming growth factor beta (TGF-β) cytokine trap having the sequence of SEQ ID NO: 14;
wherein the components of (a) and (b) are connected by a peptide linker and the cancer is colorectal cancer, head and neck cancer, or ovarian cancer.
2 . The method of claim 1 , wherein the peptide linker comprises the sequence of any one of SEQ ID NOs: 17-34.
3 . The method of claim 1 , wherein the antibody is an immunoglobulin G (IgG) antibody.
4 . The method of claim 3 , wherein the IgG antibody comprises a mutation at position 108 of SEQ ID NO: 146 or 292.
5 . The method of claim 4 , wherein the mutation is S108P mutation.
6 . The method of claim 1 , wherein the peptide linker connects the VH region with the TGF-β cytokine trap.
7 . The method of claim 1 , wherein the antibody, or antigen-binding fragment or variant thereof, comprises
a VH region having the sequence of SEQ ID NO: 6 and a VL region having the sequence of SEQ ID NO: 12.
8 . The method of claim 1 , wherein the antibody, or antigen-binding fragment or variant thereof, comprises
a VH region having the sequence of SEQ ID NO: 7 and a VL region having the sequence of SEQ ID NO: 13.
9 . The method of claim 1 , wherein the subject is a human.
10 . The method of claim 1 , further comprising administering to the subject an effective amount of T cells engineered to express an exogenous receptor.
11 . The method of claim 10 , wherein the exogenous receptor is a chimeric antigen receptor comprising an antigen-binding domain that binds to an epitope on CD19, BCMA, CD44, α-Folate receptor, CAIX, CD30, ROR1, CEA, EGP-2, EGP-40, HER2, HER3, Folate-binding Protein, GD2, GD3, IL-13R-a2, KDR, EDB-F, mesothelin, CD22, EGFR, Folate receptor α, MUC-1, MUC-4, MUC-16, MAGE-A1, h5T4, PSMA, TAG-72, EGFR, CD20, EGFRVIII, CD123, or VEGF-R2.
12 . The method of claim 10 , wherein the exogenous receptor is a chimeric antigen receptor and the engineered T cells further express a fusion protein comprising IL-15 and IL-15Rα.
13 . The method of claim 1 , wherein the fusion protein comprises either:
(a) the amino acid sequences of SEQ ID NO: 15 and SEQ ID NO: 294; or
(b) the amino acid sequences of SEQ ID NO: 296 and SEQ ID NO: 144.
14 . The method of claim 1 , wherein the cancer is colorectal cancer.
15 . The method of claim 11 , wherein the chimeric antigen receptor comprises an antigen-binding domain that binds to an epitope on CD33.
16 . The method of claim 1 , wherein the fusion protein comprises either:
(a) (i) the amino acid sequence of SEQ ID NO: 15, and (ii) the sequence of SEQ ID NO: 16, 143, or 294; or (b) (i) the amino acid sequence of SEQ ID NO: 296, and (ii) the amino acid sequence of SEQ ID NO: 144, 145, or 295.
17 . The method of claim 1 , wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 296 and the amino acid sequence of SEQ ID NO: 144.