Fusion protein comprising anti-CD3 single domain antibody, exosomal protein CD63 and RNA binding protein, and uses thereof
The present disclosure provides a fusion protein including an anti-CD3 single domain antibody, an exosomal protein CD63 and an RNA binding protein. The fusion protein of the present disclosure achieves the effect of treating cancer, immunoregulation and activating immune cells through various efficacy experiments.
1 . A fusion protein, comprising an anti-CD3 single domain antibody, an exosomal protein, and an RNA binding protein, wherein the amino acid sequence of the anti-CD3 single domain antibody is a heavy chain variable domain (VHH) sequence comprising SEQ ID NO: 2, and the exosomal protein is CD63.
2 . The fusion protein according to claim 1 , wherein the anti-CD3 single domain antibody specifically binds to a CD3 ε.
3 . The fusion protein according to claim 1 , wherein the anti-CD3 single domain antibody is an anti-T cell single domain antibody.
4 . The fusion protein according to claim 1 , wherein the amino acid sequence of N-terminus of the CD63 is SEQ ID NO:3, and the amino acid sequence of C-terminus of the CD63 is SEQ ID NO:4.
5 . The fusion protein according to claim 1 , wherein the RNA binding protein is heterogeneous nuclear ribonucleoprotein (hnRNP) A2B1.
6 . The fusion protein according to claim 5 , wherein the hnRNP A2B1 comprises an amino acid sequence of SEQ ID NO:5.
7 . The fusion protein according to claim 1 , comprising an amino acid sequence of SEQ ID NO:1.
8 . The fusion protein according to claim 1 , further comprising an extracellular vesicle (EV) sorting motif.
9 . The fusion protein according to claim 8 , wherein a nucleotide sequence encoding the amino acid sequence of the EV sorting motif is SEQ ID NO:6.
10 . The fusion protein according to claim 8 , further comprising an HLA-G chimeric antigen receptor (CAR).
11 . A pharmaceutical composition, comprising the fusion protein according to claim 1 and a pharmaceutically acceptable carrier.
12 . The pharmaceutical composition according to claim 11 , wherein the anti-CD3 single domain antibody specifically binds to a CD3 ε.
13 . The pharmaceutical composition according to claim 11 , wherein the anti-CD3 single domain antibody is an anti-T cell single domain antibody.
14 . The pharmaceutical composition according to claim 11 , wherein the amino acid sequence of N-terminus of the CD63 is SEQ ID NO:3, and the amino acid sequence of C-terminus of the CD63 is SEQ ID NO:4.
15 . The pharmaceutical composition according to claim 11 , wherein the RNA binding protein is heterogeneous nuclear ribonucleoprotein (hnRNP) A2B1.
16 . A method for treating cancer, immunoregulation and activating immune cells, comprising administering to a subject in need thereof the pharmaceutical composition according to claim 11 .
17 . The method according to claim 16 , wherein the fusion protein enhances anti-tumor activity of peripheral blood mononuclear cells (PBMCs).
18 . The method according to claim 16 , wherein the cancer is colorectal cancer, lung adenocarcinoma, glioblastoma, or pancreatic cancer.