Disrupting tumor tissues by targeting fibroblast activation protein (FAP)

1. A nucleic acid comprising a polynucleotide sequence encoding a chimeric antigen receptor (CAR) capable of binding FAP, comprising an antigen-binding domain, a transmembrane domain, and an intracellular domain, wherein the antigen-binding domain comprises:

a heavy chain variable region that comprises three heavy chain complementarity determining regions (HCDRs), wherein HCDR1 comprises the amino acid sequence YTITSYSLH (SEQ ID NO: 1), HCDR2 comprises the amino acid sequence EINPANGDHNFSEKFEIK (SEQ ID NO: 2), and HCDR3 comprises the amino acid sequence LDDSRFHWYFDV (SEQ ID NO: 3); and

a light chain variable region that comprises three light chain complementarity determining regions (LCDRs), wherein LCDR1 comprises the amino acid sequence TASSSVSYMY (SEQ ID NO: 4), LCDR2 comprises the amino acid sequence LTSNLA (SEQ ID NO: 5), and LCDR3 comprises the amino acid sequence QQWSGYPPIT (SEQ ID NO: 6).

2. The nucleic acid of claim 1 , wherein the CAR comprises a polypeptide selected from the group consisting of:

(a) a heavy chain variable region encoded by a polynucleotide sequence at least 80% identical to SEQ ID NO: 8;

(b) a light chain variable region encoded by a polynucleotide sequence at least 80% identical to SEQ ID NO: 10;

(c) a heavy chain variable region encoded by a polynucleotide sequence at least 80% identical to SEQ ID NO: 8; and a light chain variable region encoded by a polynucleotide sequence at least 80% identical to SEQ ID NO: 10;

(d) a single-chain variable fragment (scFv) encoded by a polynucleotide sequence at least 80% identical to SEQ ID NO: 12; and

(e) a single-chain variable fragment (scFv) encoded by a polynucleotide sequence at least 80% identical to SEQ ID NO: 14.

3. The nucleic acid of claim 1 , wherein:

(a) the transmembrane domain comprises a transmembrane domain of CD8 alpha; and/or

(b) the intracellular domain comprises one or more domains selected from the group consisting of:

(i) a costimulatory signaling domain and an intracellular signaling domain;

(ii) a costimulatory signaling domain comprising a costimulatory domain of 4-1BB;

(iii) a costimulatory domain of DAP12;

(iv) a costimulatory domain of CD28;

(v) a costimulatory domain of 4-1BB and a costimulatory domain of CD28;

(vi) a costimulatory domain of DAP12 and a costimulatory domain of CD28; and

(vii) an intracellular domain of CD34.

4. A nucleic acid comprising a polynucleotide sequence at least 80% identical to the sequence selected from the group consisting of SEQ ID NO: 22 and 24.

5. A vector comprising the nucleic acid of claim 1 .

6. The vector of claim 5 , wherein the vector is an expression vector and/or the vector is selected from the group consisting of a DNA vector, an RNA vector, a plasmid, a lentiviral vector, an adenoviral vector, an adeno-associated viral vector, and a retroviral vector.

7. A modified cell comprising the nucleic acid of claim 1 , wherein the modified cell is selected from the group consisting of an immune cell and a precursor cell thereof.

8. A modified cell comprising a chimeric antigen receptor (CAR) capable of binding fibroblast activation protein (FAP), wherein the modified cell is selected from the group consisting of an immune cell and a precursor cell thereof, and wherein the CAR comprises:

a heavy chain variable region that comprises three heavy chain complementarity determining regions (HCDRs), wherein HCDR1 comprises the amino acid sequence YTITSYSLH (SEQ ID NO: 1), HCDR2 comprises the amino acid sequence EINPANGDHNFSEKFEIK (SEQ ID NO: 2), and HCDR3 comprises the amino acid sequence LDDSRFHWYFDV (SEQ ID NO: 3); and

a light chain variable region that comprises three light chain complementarity determining regions (LCDRs), wherein LCDR1 comprises the amino acid sequence TASSSVSYMY (SEQ ID NO: 4), a LCDR2 comprises the amino acid sequence LTSNLA (SEQ ID NO: 5), and LCDR3 comprises the amino acid sequence QQWSGYPPIT (SEQ ID NO: 6).

9. The modified cell of claim 8 , wherein the CAR is selected from the group consisting of:

(a) a CAR comprising a heavy chain variable region comprising an amino acid sequence at least 80% identical to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence at least 80% identical to SEQ ID NO: 9;

(b) a CAR comprising a single-chain variable fragment (scFv) comprising an amino acid sequence at least 80% identical to SEQ ID NO: 11;

(c) a CAR comprising a single-chain variable fragment (scFv) comprising an amino acid sequence at least 80% identical to SEQ ID NO: 13;

(d) a CAR comprising an amino acid sequence at least 80%, identical to SEQ ID NO: 23; and

(e) a CAR comprising an amino acid sequence at least 80% identical to SEQ ID NO: 25.

10. The modified cell of claim 8 , wherein the CAR is capable of binding human FAP.

11. The modified cell of claim 8 , wherein the modified cell is selected from the group consisting of:

(a) a modified T cell;

(b) a modified NK cell;

(c) an autologous cell;

(d) an autologous cell obtained from a human subject;

(e) an autologous cell obtained from a canine subject; and

(f) an allogeneic cell.

12. A pharmaceutical composition comprising a therapeutically effective amount of the modified cell of claim 8 , and a pharmaceutically acceptable excipient.