T CELL RECEPTORS TARGETING TP53 HOTSPOT MUTATIONS AND USES THEREOF

1 . A recombinant T cell receptor (TCR) comprising an extracellular domain that binds to a TP53 peptide, wherein the TP53 peptide comprises a R175 mutation,

wherein the extracellular domain comprises an α chain comprising an α chain variable region and α chain constant region, and a β chain comprising a β chain variable region and a β chain constant region, wherein

a) the α chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 1, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 3; and the β chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 4, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 5, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 6;

b) the α chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 11, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 12, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 13; and the β chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 14, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 15, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 16;

c) the α chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 21, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 22, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 23; and the β chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 24, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 25, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 26;

d) the α chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 31, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 32, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 33; and the β chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 34, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 35, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 36;

e) the α chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the β chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46;

f) the α chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 51, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 52, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 53; and the β chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 54, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 55, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 56;

g) the α chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 61, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 62, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 63; and the β chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 64, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 65, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 66;

h) the α chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 71, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 72, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 73; and the β chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 75, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 76;

i) the α chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 81, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 82, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 83; and the β chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 84, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 85, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 86;

j) the α chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 91, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 92, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 93; and the β chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 94, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 95, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 96;

k) the α chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 101, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 102, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 103; and the β chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 104, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 105, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 106; or

l) the α chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 111, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 112, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 113; and the β chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 114, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 115, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 116.

2 . The recombinant TCR of claim 1 , wherein the α chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 81, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 82, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 83; and the β chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 84, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 85, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 86.

3 . The recombinant TCR of claim 1 , wherein the TP53 peptide comprises a R175H mutation.

4 . The recombinant TCR of claim 1 , wherein the TP53 peptide comprises or consists of the amino acid sequence set forth in SEQ ID NO: 146 or SEQ ID NO: 147.

5 . The recombinant TCR of claim 1 , wherein the TP53 peptide is associated with an HLA class I complex.

6 . The recombinant TCR of claim 1 , wherein the HLA class I complex is an HLA-A.

7 . The recombinant TCR of claim 1 , wherein the HLA-A is an HLA-A*02 superfamily member selected from the group comprising HLA-A*02:02, HLA-A*02:03, HLA-A*02:04, HLA-A*02:05, HLA-A*02:06, HLA-A*02:07, HLA-A*02:09, HLA-A*02:11, HLA-A*02:12, HLA-A*02:13, HLA-A*02:0215HLA-A*02:14, HLA-A*02:17, HLA-A*02:71, HLA-A*02:77, HLA-A*68:02, and HLA-A*69:01.

8 . The recombinant TCR of claim 1 , wherein:

a) the α chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 7, and the β chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 8;

b) the α chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 17, and the β chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 18;

c) the α chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 27, and the β chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 28;

d) the α chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 37, and the β chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 38;

e) the α chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 47, and the β chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 48;

f) the α chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 57, and the β chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 58;

g) the α chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 67, and the β chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 68;

h) the α chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 77, and the β chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 78;

i) the α chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 87, and the β chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 88;

j) the α chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 97, and the β chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 98;

k) the α chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 107, and the β chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 108; or

l) the α chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 117, and the β chain variable region comprises an amino acid sequence that is at least about 80% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 118.

9 . The recombinant TCR of claim 1 , wherein:

a) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 7, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 8;

b) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 17, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 18;

c) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 27, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 28;

d) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 37, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 38;

e) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 47, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 48;

f) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 57, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 58;

g) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 67, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 68;

h) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 77, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 78;

i) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 87, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 88;

j) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 97, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 98;

k) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 107, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 108; or

l) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 117, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 118.

10 . The recombinant TCR of claim 1 , wherein:

a) the α chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 9, and the β chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 10;

b) the α chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 19, and the β chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 20;

c) the α chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 29, and the β chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 30;

d) the α chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 39, and the β chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 40;

e) the α chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 49, and the β chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 50;

f) the α chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 59, and the β chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 60;

g) the α chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 69, and the β chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 70;

h) the α chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 79, and the β chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 80;

i) the α chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 89, and the β chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 90;

j) the α chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 99, and the β chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 100;

k) the α chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 109, and the β chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 110; or

l) the α chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 119, and the β chain comprises an amino acid sequence that is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 120.

11 . The recombinant TCR of claim 1 , wherein:

a) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 9, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 10;

b) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 19, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 20;

c) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 29, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 30;

d) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 39, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 40;

e) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 49, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 50;

f) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 59, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 60;

g) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 69, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 70;

h) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 79, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 80;

i) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 89, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 90;

j) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 99, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 100;

k) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 109, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 110; or

l) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 119, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 120.

12 . An immunoresponsive cell comprising the TCR of claim 1 .

13 . The cell of claim 12 , wherein the cell is selected from the group consisting of a T cell, a Natural Killer (NK) cell, and a pluripotent stem cell from which a lymphoid cell may be differentiated.

14 . The cell of claim 13 , wherein the cell is a T cell or an NK cell.

15 . The cell of claim 14 , wherein the T cell is selected from the group consisting of a cytotoxic T lymphocyte (CTL), a regulatory T cell, a γδ T cell, a Natural Killer-T cell (NK-T), a stem cell memory T cell, a central memory T cell, and an effector memory T cell.

16 . A composition comprising the cell of claim 12 .

17 . The composition of claim 16 , which is a pharmaceutical composition further comprising a pharmaceutically acceptable carrier.

18 . A nucleic acid encoding the T cell receptor (TCR) of claim 1 .

19 . A vector comprising the nucleic acid of claim 18 .

20 . The vector of claim 19 , wherein (a) the vector is a γ-retroviral vector, or (b) the vector is a plasmid comprising a backbone comprising fewer than about 500 nucleotides.

21 . A lipid nanoparticle comprising the nucleic acid of claim 18 .

22 . A method for producing a cell that binds to a TP53 peptide that comprises a R175 mutation, comprising introducing into the cell the nucleic acid of claim 18 .

23 . A method of treating and/or preventing a tumor or a premalignant condition associated with TP53 in a subject, comprising administering to the subject an effective amount of the cell of claim 12 or a composition thereof.

24 . The method of claim 23 , wherein the TP53 mutation is a R175H mutation.

25 . The method of claim 23 , wherein the tumor is selected from the group consisting of brain tumors, soft tissue and bone sarcomas, adrenocortical carcinoma, rhabdomyosarcoma, Wilms tumor, pancreatic cancer, breast cancer, endometrial cancer, cervical cancer, anal cancer, bladder cancer, colorectal cancer, cholangiocarcinoma/bile duct cancer, lung cancer, ovarian cancer, esophageal cancer, gastric cancer, head and neck squamous cell carcinoma, nonmelanoma skin cancer, salivary gland cancer, melanoma, multiple myeloma, prostate cancer, T-lymphoblastic leukemia/lymphoma, acute myeloid leukemia, B-cell leukemia, myeloproliferative neoplasm, and histiocytosis.

26 . The method of claim 23 , wherein the premalignant condition is clonal hematopoiesis.

27 . The method of claim 23 , wherein the subject is a human.

28 . The method of claim 27 , wherein the subject comprises an HLA-A.

29 . The method of claim 28 , wherein the HLA-A is an HLA-A*02 superfamily member selected from the group comprising HLA-A*02, HLA-A*02:01, HLA-A*02:02, HLA-A*02:03, HLA-A*02:04, HLA-A*02:05, HLA-A*02:06, HLA-A*02:07, HLA-A*02:09, HLA-A*02:11, HLA-A*02:12, HLA-A*02:13, HLA-A*02:0215HLA-A*02:14, HLA-A*02:17, HLA-A*02:71, HLA-A*02:77, HLA-A*68:02, and HLA-A*69:01.

30 . The method of claim 29 , wherein the HLA-A*02 superfamily member is HLA-A*02:01.