Anti-glypican-1-immunizing antigen receptor
The purpose of the present invention is to produce a chimeric antigen receptor (CAR) specific to glypican-1 (GPC-1) and to treat squamous cell carcinoma with genetically modified cells capable of expressing the CAR. The present invention provides: a chimeric antigen receptor for use in the treatment and/or prevention of squamous cell carcinoma, said chimeric antigen receptor comprising an extracellular domain capable of binding to GPC-1, a transmembrane domain and one or multiple intracellular domains, wherein at least one of the intracellular domains is an intracellular domain containing a primary cytosolic signaling sequence or an intracellular domain containing both a primary cytosolic signaling sequence and a secondary cytosolic signaling sequence; a genetically modified cell capable of expressing the chimeric antigen receptor; and a cell preparation containing the cell.
1 . A method for treatment and/or prevention of a solid tumor expressing GPC-1, comprising administering a CAR-T cell comprising a nucleic acid to an individual in need thereof, wherein the nucleic acid comprises an extracellular domain capable of binding to glypican-1 (GPC-1), a transmembrane domain and one or more intracellular domains, wherein at least one of the intracellular domains is an intracellular domain comprising a primary cytosolic signaling sequence; wherein the extracellular domain capable of binding to GPC-1 comprises the heavy chain variable region (VH) and light chain variable region (VL) of anti-GPC-1 antibody and wherein the nucleotide sequence encoding the heavy chain variable region of anti-GPC-1 antibody comprises the nucleotide sequence listed as SEQ ID NO: I or a nucleotide sequence with at least 95% identity therewith and having the same function, and the nucleotide sequence encoding the light chain variable region comprises the nucleotide sequence listed as SEQ ID NO: 2 or a nucleotide sequence with at least 95% identity therewith and having the same function.
2 . The method according to claim 1 , wherein the solid tumor is squamous cell carcinoma, esophageal cancer or colon cancer.
3 . A vector containing a nucleic acid, wherein the nucleic acid comprises an extracellular domain capable of binding to glypican-1 (GPC-1), a transmembrane domain and one or more intracellular domains, wherein at least one of the intracellular domains is an intracellular domain comprising a primary cytosolic signaling sequence; wherein the extracellular domain capable of binding to GPC-1 comprises the heavy chain variable region (VH) and light chain variable region (VL) of anti-GPC-1 antibody and wherein the nucleotide sequence encoding the heavy chain variable region of anti-GPC-1 antibody comprises the nucleotide sequence listed as SEQ ID NO: I or a nucleotide sequence with at least 95% identity therewith and having the same function, and the nucleotide sequence encoding the light chain variable region comprises the nucleotide sequence listed as SEQ ID NO: 2 or a nucleotide sequence with at least 95% identity therewith and having the same function.
4 . Cells comprising the vector of claim 3 , wherein the cells express a chimeric antigen receptor.
5 . The cells according to claim 4 , wherein the cells are T cells or a cell population comprising T cells.
6 . A cell preparation containing cells according to claim 5 , for treatment of a solid tumor expressing GPC-1.
7 . The cell preparation according to claim 6 , wherein the solid tumor is squamous cell carcinoma, esophageal cancer or colon cancer.
8 . A medical composition comprising a nucleic acid and a medically acceptable excipient, wherein the nucleic acid comprises an extracellular domain capable of binding to glypican-1 (GPC-1), a transmembrane domain and one or more intracellular domains, wherein at least one of the intracellular domains is an intracellular domain comprising a primary cytosolic signaling sequence; wherein the extracellular domain capable of binding to GPC-1 comprises the heavy chain variable region (VH) and light chain variable region (VL) of anti-GPC-1 antibody and wherein the nucleotide sequence encoding the heavy chain variable region of anti-GPC-1 antibody comprises the nucleotide sequence listed as SEQ ID NO: I or a nucleotide sequence with at least 95% identity therewith and having the same function, and the nucleotide sequence encoding the light chain variable region comprises the nucleotide sequence listed as SEQ ID NO: 2 or a nucleotide sequence with at least 95% identity therewith and having the same function.
9 . A medical composition according to claim 8 , for treatment of a solid tumor expressing GPC-1.
10 . A medical composition according to claim 9 , wherein the solid tumor is squamous cell carcinoma, esophageal cancer or colon cancer.
11 . A method for treatment of a solid tumor expressing GPC-1 in a subject, wherein the method comprising administering to the subject a nucleic acid that encodes a polypeptide comprising an extracellular domain capable of binding to glypican-1 (GPC-1), a transmembrane domain and one or more intracellular domains, wherein at least one of the intracellular domains is an intracellular domain comprising a primary cytosolic signaling sequence; wherein the extracellular domain capable of binding to GPC-1 comprises the heavy chain variable region (VH) and light chain variable region (VL) of anti-GPC-1 antibody and wherein the nucleotide sequence encoding the heavy chain variable region of anti-GPC-1 antibody comprises the nucleotide sequence listed as SEQ ID NO: I or a nucleotide sequence with at least 95% identity therewith and having the same function, and the nucleotide sequence encoding the light chain variable region comprises the nucleotide sequence listed as SEQ ID NO: 2 or a nucleotide sequence with at least 95% identity therewith and having the same function.
12 . The method according to claim 11 , wherein the solid tumor is squamous cell carcinoma, esophageal cancer or colon cancer.