Modified T cell receptors and related materials and methods
View Patent ↗The invention is directed to a modified T cell receptor (TCR) comprising an amino acid sequence of a wild-type (WT) TCR with no more than three amino acid substitutions, wherein the modified TCR, as compared to the WT TCR, (i) has an enhanced ability to recognize target cells when expressed by CD4 + T cells and (ii) does not exhibit a decrease in antigen specificity when expressed by CD8 + T cells. Polypeptides, proteins, nucleic acids, recombinant expression vectors, host cells, populations of cells, antibodies, and pharmaceutical compositions related to the modified TCR also are part of the invention. Further, the invention is directed to methods of detecting a diseased cell in a host, methods of treating or preventing a disease in a host, and methods of identifying a candidate adoptive immunotherapy TCR.
1. A modified T cell receptor (TCR) comprising an amino acid sequence of a wild-type (WT) TCR with no more than three amino acid substitutions located in a complementarity determining region (CDR) 2 of a beta chain of the TCR, wherein the modified TCR retains the antigen specificity of the WT TCR and, as compared to the WT TCR, (i) has an enhanced ability to recognize target cells when expressed by CD4 + T cells and (ii) does not exhibit a decrease in antigen specificity when expressed by CD8 + T cells, and wherein
a) the modified TCR has antigen specificity for cancer antigen MART-1, or
b) the modified TCR has antigen specificity for cancer antigen NY-ESO-1 and comprises the amino acid sequence of SEQ ID NO: 8 with no more than three amino acid substitutions located in the CDR2 of SEQ ID NO: 8.
2. The modified TCR of claim 1 , wherein the amino acid substitutions are conservative amino acid substitutions.
3. The modified TCR of claim 2 , wherein the conservative amino acid substitutions are selected from the group consisting of T→A, G→A, A→I, T→V, A→M, T→I, A→V, T→G, V→A, and T→S.
4. The modified TCR of claim 1 , wherein the modified TCR comprises the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.
5. The modified TCR of claim 4 , wherein the modified TCR comprises the amino acid sequence of SEQ ID NO: 14, SEQ ID NO: 15, or SEQ ID NO: 16.
6. The modified TCR of claim 4 , wherein the modified TCR additionally comprises the amino acid sequence of SEQ ID NO: 7.
7. The modified TCR of claim 1 , wherein the TCR has antigen specificity for MART-1.
8. The modified TCR of claim 7 , wherein the TCR comprises the amino acid sequence of SEQ ID NO: 10 or SEQ ID NO: 12 and the TCR has no more than three amino acid substitutions.
9. The modified TCR of claim 8 , wherein the modified TCR comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 4 to 6, 13, and 95 to 103.
10. The modified TCR of claim 9 , wherein the modified TCR comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 17 to 20, 37 to 39, and 104 to 109.
11. The modified TCR of claim 9 , wherein the modified TCR additionally comprises an amino acid sequence of SEQ ID NO: 9 or SEQ ID NO. 11.
12. The modified TCR of claim 11 , wherein the modified TCR comprises the amino acid sequence of:
(i) any of SEQ ID NOs: 17, 20, 37 to 39, and 104 to 109 in combination with SEQ ID NO. 9; or
(ii) SEQ ID NO: 18 or 19 in combination with SEQ ID NO: 11.
13. The modified TCR of claim 1 comprising a leader sequence selected from the group consisting of SEQ ID NOs: 110 to 115.
14. The modified TCR of claim 1 , wherein the modified TCR comprises the amino acid sequence of SEQ ID NO: 118.
15. The modified TCR of claim 14 , wherein the modified TCR comprises the amino acid sequence of SEQ ID NO: 118, wherein
Xaa at position 51 is G or A; and
Xaa at position 52 is V or A,
with the proviso that Xaa at position 51 is not G when at position 52 is V.
16. The modified TCR of claim 1 , wherein the modified TCR comprises the amino acid sequence of SEQ ID NO: 119, SEQ ID NO: 120, or SEQ ID NO: 121.
17. The modified TCR of claim 14 , wherein the modified TCR comprises the amino acid sequence of SEQ ID NO: 119, wherein
Xaa at position 51 is selected from the group consisting of T, I, A, and V;
Xaa at position 52 is selected from the group consisting of A, V, I, and M; and
Xaa at position 54 is selected from the group consisting of T, G, S, and A,
with the proviso that Xaa at position 51 is not T when Xaa at position 52 is A and Xaa at position 54 is T.
18. The modified TCR of claim 14 , wherein the modified TCR comprises the amino acid sequence of SEQ ID NO: 120, wherein
Xaa at position 51 is G or A;
Xaa at position 52 is A or I,
with the proviso that Xaa at position 51 is not G when Xaa at position 52 is A.
19. The modified TCR of claim 14 , wherein the modified TCR comprises the amino acid sequence of SEQ ID NO: 121, wherein
Xaa at position 51 is G or A; and
Xaa at position 52 is V or A,
with the proviso that Xaa at position 51 is not G when Xaa at position 52 is V.
20. An isolated polypeptide comprising a functional portion of the modified TCR of claim 1 , wherein the functional portion comprises variable regions of alpha and beta chains of the TCR and wherein the functional portion comprises the amino acid substitutions.
21. The isolated polypeptide of claim 20 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 6, 13, and 95 to 103.
22. The isolated polypeptide of claim 21 comprising an amino acid sequence selected from the group consisting of SEQ NOs: 14 to 20, 37 to 39, and 104 to 109.
23. The isolated polypeptide of claim 20 , wherein the polypeptide additionally comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 7, 9, and 11.
24. An isolated protein comprising the polypeptide of claim 20 .
25. The isolated protein of claim 24 , comprising (i) a first polypeptide chain comprising the amino acid sequence of SEQ ID NO: 1, 2, or 3 and a second polypeptide chain comprising the amino acid sequence of SEQ ID NO: 7, (ii) a first polypeptide chain comprising the amino acid sequence of any of SEQ ID NOs: 4, 13, and 95 to 103 and a second polypeptide chain comprising the amino acid sequence of SEQ ID NO: 9, or (iii) a first polypeptide chain comprising the amino acid sequence of SEQ ID NO: 5 or 6 and a second polypeptide chain comprising the amino acid sequence of SEQ ID NO: 11.
26. A pharmaceutical composition comprising the modified TCR of claim 1 .
27. A method of detecting a diseased cell in a host, wherein the diseased cell expresses an antigen characteristic of a disease, comprising:
(a) contacting a sample comprising cells of the host with the modified TCR of claim 1 , thereby forming a complex between the modified TCR and the antigen, and
(b) detecting the complex,
wherein detection of the complex is indicative of a diseased cell in the host.
28. The method of claim 27 , wherein the diseased cell is a cancer cell or an infected cell.
29. The method of claim 28 , wherein the cancer cell is a melanoma cell.
30. The method of claim 27 , wherein the host is a human.
31. The method of claim 27 , wherein the method is an in vitro method.