Human t cell receptors and uses thereof

Abstract

The present disclosure provides T cell receptors (TCRs) that are capable of binding to Wilms Tumour 1 (WT-1), (e.g., antigenic peptides derived from WT-1). Also provided are multispecific antigen-binding molecules comprising such TCRs, nucleic acids encoding such TCRs, vectors comprising such nucleic acids, cells comprising such TCRs / nucleic acids / vectors, and compositions comprising such TCRs / nucleic acids / vectors / cells. Further provided are methods for treating / preventing diseases / conditions using such TCRs, multispecific antigen-binding molecules, nucleic acids, vectors, cells and / or compositions.

Description

[0001] Human T Cell Receptors and Uses Thereof

[0002] This application claims priority from US 63 / 733736 filed 13 December 2024, the contents and elements of which are herein incorporated by reference for all purposes.

[0003] Technical Field

[0004] The present invention relates to TCR sequences, and their use in TCR-based immunotherapies and in the treatment of WT-1 -expressing neoplasms and cancers.

[0005] Background

[0006] Immunotherapy has revolutionized cancer treatment. Starting with early observations that a patient’s own T cells can kill tumor cells (Rosenberg & Restifo, 2015), ongoing research has led to the development of immune checkpoint inhibitors, adoptive transfer of ex vivo expanded tumor infiltrating T cells, CAR-T cell therapy and TCR-T therapy.

[0007] The binding domain of Chimeric antigen receptors (CARs) are based on antibody fragments that bind to unprocessed antigens, such as tumor antigens on the surface of tumor cells. Upon binding to a certain threshold number of target antigens displayed on the target cell, the CAR-expressing T cell can become activated and can kill the tumor cell.

[0008] The binding mechanism of a T cell receptor (TCR) is entirely different from that of a CAR. A TCR is a natural component of T cells. Upon binding of potentially a single TCR with a single MHC class I molecule displaying the target peptide of the TCR, the first step of the T cell activation cascade is initiated. This exquisite sensitivity of TCR-mediated killing by T cells provides an opportunity to use these cells in adoptive T cell therapy for cancer, particularly in cases where the level of expression of the target antigen may be low. T cells can be genetically modified to equip them with a TCR specifically targeting a peptide that is processed from a tumor antigen and presented on the surface of a patient’s tumor in the context of a Human Leukocyte Antigen (HLA) molecule. Alternatively, the TCRs can be used in other immunotherapeutic modalities, such as bi-specific molecules. An example of this modality is the marketed drug KIMMTRAK® that comprises a TCR specific for a gp100-derived peptide when presented in the context of HLA-A*02:01 , linked to a T cell engaging anti-CD3 binding domain.

[0009] TCR-based therapies are based on the binding of TCRs to peptides presented in the context of MHC molecules (also referred to herein as ‘HLA molecules’). For a TCR-based therapy against cancer, the target antigens of the TCR must be expressed in tumor cells, and the patient must express the HLA allele to which the TCR is restricted. As part of immune presentation of antigens to CD8+ T cells, antigens are broken down into peptides by the proteasome or immunoproteasome, leading to the presentation of these peptides on MHC molecules. CD8+ T cells, via their TCR, bind to peptides presented on a given allele of MHC class I molecules. The specific binding of T cells to peptides presented by a given MHC class I molecule is called HLA restriction of the T cell response (Murphy & Weaver (2017) Janeway’s Immunobiology 9). Once a T cell has bound to the cancer cell via the TCR:MHC+peptide interaction and the cell is activated by additional stimuli, the T cell can kill the tumor cell. There are different types of tumor antigens that can be targeted for TCR-based therapy. Ideally, the antigen should be expressed exclusively or predominantly on the tumor compared to normal healthy tissue. A key success factor for TCR-based therapies is the selection of the tumor antigen, such that as many patients as possible can benefit from a given therapy. Tumor antigens that are expressed in several tumor types and in a large proportion of patients (so-called shared tumor antigens) are therefore of high priority.

[0010] Few shared tumor antigens have been tested in TCR-T cell therapy clinical trials so far (Baulu et al., 2023, Science Advances, 9(7)). Results from ongoing clinical trials with these few antigens have been very encouraging and such TCR-T therapies are likely to be approved for clinical use in the near future. The FDA has already granted accelerated approval of TECELRA® (afamitresgene autoleucel), a MAGE- A4-targeted T cell therapy for unresectable or metastatic synovial sarcoma. Hence, there is a huge opportunity to develop novel TCR-based therapies against new cancer targets, or against previously described targets restricted to diverse HLA alleles. In particular, tumor antigens that are shared across several tumor indications will be most promising for clinical application since more patients will be eligible for treatment with a given TCR therapy.

[0011] Wilms’ Tumor Antigen 1 (WT-1) is over-expressed across several major solid tumor indications in protein levels up to 10- to 1000- fold higher compared to healthy tissues. HLA-A*02 is by far the most common HLA allele, followed by HLA-A*11 and HLA-A*24. Targeting WT-1 has been associated with a favorable safety profile. In addition, WT-1 over-expression across multiple cancer indications also means that any successful therapeutic can benefit a substantial patient population.

[0012] The present invention has been devised in light of the above considerations.

[0013] Summary

[0014] The present invention relates to T cell receptors (TCRs) that are capable of binding to WT-1 , (e.g., antigenic peptides derived from WT-1).

[0015] Provided is a T cell receptor (TCR) comprising a TCRa chain variable domain comprising a CDR3a having an amino acid sequence of SEQ ID NO:34, 10, 4, 16, 22, 28, 40 or 46, or a variant thereof in which 1 or 2 or 3 amino acids are substituted with another amino acid; in combination with a TCRp chain variable domain comprising a CDR3p having an amino acid sequence of SEQ ID NO:37, 13, 7, 19, 25, 31 , 43 or 49, or a variant thereof in which 1 or 2 or 3 amino acids are substituted with another amino acid.

[0016] In some embodiments, the TCR comprises:

[0017] (a)

[0018] (i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NO:34, and / or

[0019] (ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:37;

[0020] (b) (i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ

[0021] ID NO:10, and / or

[0022] (ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:13;

[0023] (c)

[0024] (i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ

[0025] ID NO:4, and / or

[0026] (ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:7;

[0027] (d)

[0028] (i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NO:16, and / or

[0029] (ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:19;

[0030] (e)

[0031] (i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ

[0032] ID NO:22, and / or

[0033] (ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:25;

[0034] (f)

[0035] (i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ

[0036] ID NO:28, and / or

[0037] (ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:31 ;

[0038] (g)

[0039] (i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NQ:40, and / or

[0040] (ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ

[0041] ID NO:43; or

[0042] (h)

[0043] (i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ

[0044] ID NO:46, and / or

[0045] (ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:49.

[0046] In some embodiments, the TCR comprises:

[0047] (a)

[0048] (i) a TCRa chain variable domain incorporating the following CDRs:

[0049] CDR1a having the amino acid sequence of SEQ ID NO:32

[0050] CDR2a having the amino acid sequence of SEQ ID NO:33

[0051] CDR3a having the amino acid sequence of SEQ ID NO:34, and / or

[0052] (ii) a TCRp chain variable domain incorporating the following CDRs: CDR1 p having the amino acid sequence of SEQ ID NO:35 CDR2p having the amino acid sequence of SEQ ID NO:36 CDR3p having the amino acid sequence of SEQ ID NO:37;

[0053] (b)

[0054] (i) a TCRa chain variable domain incorporating the following CDRs:

[0055] CDR1a having the amino acid sequence of SEQ ID NO:8 CDR2a having the amino acid sequence of SEQ ID NO:9 CDR3a having the amino acid sequence of SEQ ID NQ:10, and / or

[0056] (ii) a TCRp chain variable domain incorporating the following CDRs:

[0057] CDR1 p having the amino acid sequence of SEQ ID NO:11 CDR2p having the amino acid sequence of SEQ ID NO:12 CDR3p having the amino acid sequence of SEQ ID NO:13;

[0058] (c)

[0059] (i) a TCRa chain variable domain incorporating the following CDRs:

[0060] CDR1a having the amino acid sequence of SEQ ID NO:2 CDR2a having the amino acid sequence of SEQ ID NO:3 CDR3a having the amino acid sequence of SEQ ID NO:4, and / or

[0061] (ii) a TCRp chain variable domain incorporating the following CDRs:

[0062] CDR1 p having the amino acid sequence of SEQ ID NO:5 CDR2p having the amino acid sequence of SEQ ID NO:6 CDR3p having the amino acid sequence of SEQ ID NO:7;

[0063] (d)

[0064] (i) a TCRa chain variable domain incorporating the following CDRs:

[0065] CDR1a having the amino acid sequence of SEQ ID NO:14 CDR2a having the amino acid sequence of SEQ ID NO:15 CDR3a having the amino acid sequence of SEQ ID NO:16, and / or

[0066] (ii) a TCRp chain variable domain incorporating the following CDRs:

[0067] CDR1 p having the amino acid sequence of SEQ ID NO:17 CDR2p having the amino acid sequence of SEQ ID NO:18 CDR3p having the amino acid sequence of SEQ ID NO:19;

[0068] (e)

[0069] (i) a TCRa chain variable domain incorporating the following CDRs:

[0070] CDR1a having the amino acid sequence of SEQ ID NQ:20 CDR2a having the amino acid sequence of SEQ ID NO:21 CDR3a having the amino acid sequence of SEQ ID NO:22, and / or

[0071] (ii) a TCRp chain variable domain incorporating the following CDRs:

[0072] CDR1 p having the amino acid sequence of SEQ ID NO:23 CDR2p having the amino acid sequence of SEQ ID NO:24 CDR3p having the amino acid sequence of SEQ ID NO:25;

[0073] (f)

[0074] (i) a TCRa chain variable domain incorporating the following CDRs: CDR1a having the amino acid sequence of SEQ ID NO:26 CDR2a having the amino acid sequence of SEQ ID NO:27

[0075] CDR3a having the amino acid sequence of SEQ ID NO:28, and / or

[0076] (ii) a TCRp chain variable domain incorporating the following CDRs:

[0077] CDR1 p having the amino acid sequence of SEQ ID NO:29 CDR2p having the amino acid sequence of SEQ ID NQ:30 CDR3p having the amino acid sequence of SEQ ID NO:31 ;

[0078] (g)

[0079] (i) a TCRa chain variable domain incorporating the following CDRs:

[0080] CDR1a having the amino acid sequence of SEQ ID NO:38 CDR2a having the amino acid sequence of SEQ ID NO:39 CDR3a having the amino acid sequence of SEQ ID NQ:40, and / or

[0081] (ii) a TCRp chain variable domain incorporating the following CDRs:

[0082] CDR1 p having the amino acid sequence of SEQ ID NO:41 CDR2p having the amino acid sequence of SEQ ID NO:42 CDR3p having the amino acid sequence of SEQ ID NO:43; or

[0083] (h)

[0084] (i) a TCRa chain variable domain incorporating the following CDRs:

[0085] CDR1a having the amino acid sequence of SEQ ID NO:44 CDR2a having the amino acid sequence of SEQ ID NO:45 CDR3a having the amino acid sequence of SEQ ID NO:46, and / or

[0086] (ii) a TCRp chain variable domain incorporating the following CDRs:

[0087] CDR1 p having the amino acid sequence of SEQ ID NO:47 CDR2p having the amino acid sequence of SEQ ID NO:48 CDR3p having the amino acid sequence of SEQ ID NO:49.

[0088] In some embodiments, the TCR comprises a TCRa chain variable domain comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NQ:60, 52, 50,

[0089] 54, 56, 58, 62 or 64.

[0090] In some embodiments, the TCR comprises a TCRp chain variable domain comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:61 , 53, 51 ,

[0091] 55, 57, 59, 63 or 65.

[0092] In some embodiments, the TCR is capable of binding to VX1DFX2X3PX4X5 (SEQ ID NO:181), VLDFX1PPX2X3 (SEQ ID NO:179), VLDFX1PX2GX3 (SEQ ID NO:182), and / or VLDFX1X2PX3X4 (SEQ ID NQ:180) when presented by an MHC class I molecule.

[0093] In some embodiments, the TCR is capable of binding to VX1DFX2X3PX4X5 (SEQ ID NQ:181)-HLA-A*02 complex, VLDFX1PPX2X3 (SEQ ID NQ:179)-HLA-A*02 complex, VLDFX1PX2GX3 (SEQ ID NO:182)-HLA- A*02 complex, and / or VLDFX1X2PX3X4 (SEQ ID NQ:180)-HLA-A*02 complex. In some embodiments, the TCR is capable of binding to VX1DFX2X3PX4X5 (SEQ ID NO:181)-HLA- A*02:01 complex, VLDFX1PPX2X3 (SEQ ID NQ:179)-HLA-A*02:01 complex, VLDFX1PX2GX3 (SEQ ID NQ:182)-HLA-A*02:01 complex, and / or VLDFX1X2PX3X4 (SEQ ID NQ:180)-HLA-A*02:01 complex.

[0094] In some embodiments, the TCR is capable of binding to VLDFAPPGA (SEQ ID NO:1) when presented by an MHC class I molecule.

[0095] In some embodiments, the TCR is capable of binding to a VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02 complex.

[0096] In some embodiments, the TCR is capable of binding to a VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02:01 complex, a VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02:07 complex, and / or a VLDFAPPGA (SEQ ID NO:1)- HLA-A*02:02 complex.

[0097] Also provided is a T cell receptor (TCR) comprising a TCRa chain variable domain and a TCRp chain variable domain, wherein the TCR is capable of binding to VX1DFX2X3PX4X5 (SEQ ID NO:181), VLDFX1PPX2X3 (SEQ ID NO:179), VLDFX1PX2GX3 (SEQ ID NO:182), and / or VLDFX1X2PX3X4 (SEQ ID NQ:180), when presented by an MHC class I molecule.

[0098] In some embodiments, the TCR is capable of binding to VX1DFX2X3PX4X5 (SEQ ID NQ:181)-HLA-A*02 complex, VLDFX1PPX2X3 (SEQ ID NQ:179)-HLA-A*02 complex, VLDFX1PX2GX3 (SEQ ID NO:182)-HLA- A*02 complex, and / or VLDFX1X2PX3X4 (SEQ ID NQ:180)-HLA-A*02 complex.

[0099] In some embodiments, the TCR is capable of binding to VX1DFX2X3PX4X5 (SEQ ID NO:181)-HLA- A*02:01 complex, VLDFX1PPX2X3 (SEQ ID NQ:179)-HLA-A*02:01 complex, VLDFX1PX2GX3 (SEQ ID NQ:182)-HLA-A*02:01 complex, and / or VLDFX1X2PX3X4 (SEQ ID NQ:180)-HLA-A*02:01 complex.

[0100] In some embodiments, the TCR is capable of binding to VLDFAPPGA (SEQ ID NO:1) when presented by an MHC class I molecule.

[0101] In some embodiments, the TCR is capable of binding to VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02 complex.

[0102] In some embodiments, the TCR is capable of binding to VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02:01 complex, a VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02:07 complex, and / or a VLDFAPPGA (SEQ ID NO:1)- HLA-A*02:02 complex.

[0103] In some embodiments, the TCR comprises a TCRa chain variable domain comprising a CDR3a having an amino acid sequence of SEQ ID NO:34, 10, 4, 16, 22, 28, 40 or 46, or a variant thereof in which 1 or 2 or 3 amino acids are substituted with another amino acid; in combination with a TCRp chain variable domain comprising a CDR3p having an amino acid sequence of SEQ ID NO:37, 13, 7, 19, 25, 31 , 43 or 49, or a variant thereof in which 1 or 2 or 3 amino acids are substituted with another amino acid, respectively.

[0104] In some embodiments, the TCR comprises:

[0105] (a)

[0106] (i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ

[0107] ID NO:34, and / or

[0108] (ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:37;

[0109] (b)

[0110] (i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ

[0111] ID NQ:10, and / or

[0112] (ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:13;

[0113] (c)

[0114] (i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ

[0115] ID NO:4, and / or

[0116] (ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NOT;

[0117] (d)

[0118] (i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ

[0119] ID NO:16, and / or

[0120] (ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:19;

[0121] (e)

[0122] (i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NO:22, and / or

[0123] (ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:25;

[0124] (f)

[0125] (i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ

[0126] ID NO:28, and / or

[0127] (ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:31 ;

[0128] (g)

[0129] (i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ

[0130] ID NQ:40, and / or

[0131] (ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ

[0132] ID NO:43; or

[0133] (h)

[0134] (i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ

[0135] ID NO:46, and / or (ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:49.

[0136] In some embodiments, the TCR comprises:

[0137] (a)

[0138] (i) a TCRa chain variable domain incorporating the following CDRs:

[0139] CDR1a having the amino acid sequence of SEQ ID NO:32 CDR2a having the amino acid sequence of SEQ ID NO:33

[0140] CDR3a having the amino acid sequence of SEQ ID NO:34, and / or

[0141] (ii) a TCRp chain variable domain incorporating the following CDRs:

[0142] CDR1 p having the amino acid sequence of SEQ ID NO:35 CDR2p having the amino acid sequence of SEQ ID NO:36 CDR3p having the amino acid sequence of SEQ ID NO:37;

[0143] (b)

[0144] (i) a TCRa chain variable domain incorporating the following CDRs:

[0145] CDR1a having the amino acid sequence of SEQ ID NO:8 CDR2a having the amino acid sequence of SEQ ID NO:9 CDR3a having the amino acid sequence of SEQ ID NQ:10, and / or

[0146] (ii) a TCRp chain variable domain incorporating the following CDRs:

[0147] CDR1 p having the amino acid sequence of SEQ ID NO:11 CDR2p having the amino acid sequence of SEQ ID NO:12 CDR3p having the amino acid sequence of SEQ ID NO:13;

[0148] (c)

[0149] (i) a TCRa chain variable domain incorporating the following CDRs:

[0150] CDR1a having the amino acid sequence of SEQ ID NO:2 CDR2a having the amino acid sequence of SEQ ID NO:3 CDR3a having the amino acid sequence of SEQ ID NO:4, and / or

[0151] (ii) a TCRp chain variable domain incorporating the following CDRs:

[0152] CDR1 p having the amino acid sequence of SEQ ID NO:5 CDR2p having the amino acid sequence of SEQ ID NO:6

[0153] CDR3p having the amino acid sequence of SEQ ID NO:7;

[0154] (d)

[0155] (i) a TCRa chain variable domain incorporating the following CDRs:

[0156] CDR1a having the amino acid sequence of SEQ ID NO:14 CDR2a having the amino acid sequence of SEQ ID NO:15 CDR3a having the amino acid sequence of SEQ ID NO:16, and / or

[0157] (ii) a TCRp chain variable domain incorporating the following CDRs:

[0158] CDR1 p having the amino acid sequence of SEQ ID NO:17 CDR2p having the amino acid sequence of SEQ ID NO:18 CDR3p having the amino acid sequence of SEQ ID NO:19;

[0159] (e)

[0160] (i) a TCRa chain variable domain incorporating the following CDRs: CDR1a having the amino acid sequence of SEQ ID NO:20

[0161] CDR2a having the amino acid sequence of SEQ ID NO:21

[0162] CDR3a having the amino acid sequence of SEQ ID NO:22, and / or

[0163] (ii) a TCRp chain variable domain incorporating the following CDRs:

[0164] CDR1 p having the amino acid sequence of SEQ ID NO:23 CDR2p having the amino acid sequence of SEQ ID NO:24 CDR3p having the amino acid sequence of SEQ ID NO:25;

[0165] (f)

[0166] (i) a TCRa chain variable domain incorporating the following CDRs:

[0167] CDR1a having the amino acid sequence of SEQ ID NO:26 CDR2a having the amino acid sequence of SEQ ID NO:27

[0168] CDR3a having the amino acid sequence of SEQ ID NO:28, and / or

[0169] (ii) a TCRp chain variable domain incorporating the following CDRs:

[0170] CDR1 p having the amino acid sequence of SEQ ID NO:29 CDR2p having the amino acid sequence of SEQ ID NQ:30 CDR3p having the amino acid sequence of SEQ ID NO:31 ;

[0171] (g)

[0172] (i) a TCRa chain variable domain incorporating the following CDRs:

[0173] CDR1a having the amino acid sequence of SEQ ID NO:38 CDR2a having the amino acid sequence of SEQ ID NO:39

[0174] CDR3a having the amino acid sequence of SEQ ID NQ:40, and / or

[0175] (ii) a TCRp chain variable domain incorporating the following CDRs:

[0176] CDR1 p having the amino acid sequence of SEQ ID NO:41 CDR2p having the amino acid sequence of SEQ ID NO:42 CDR3p having the amino acid sequence of SEQ ID NO:43; or

[0177] (h)

[0178] (i) a TCRa chain variable domain incorporating the following CDRs:

[0179] CDR1a having the amino acid sequence of SEQ ID NO:44 CDR2a having the amino acid sequence of SEQ ID NO:45 CDR3a having the amino acid sequence of SEQ ID NO:46, and / or

[0180] (ii) a TCRp chain variable domain incorporating the following CDRs:

[0181] CDR1 p having the amino acid sequence of SEQ ID NO:47 CDR2p having the amino acid sequence of SEQ ID NO:48 CDR3p having the amino acid sequence of SEQ ID NO:49.

[0182] In some embodiments, the TCR comprises a TCRa chain variable domain comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NQ:60, 52, 50,

[0183] 54, 56, 58, 62 or 64.

[0184] In some embodiments, the TCR comprises a TCRp chain variable domain comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:61 , 53, 51 ,

[0185] 55, 57, 59, 63 or 65. In some embodiments, the TCR comprises a TCRa chain variable domain encoded by a nucleotide sequence having at least 80% sequence identity to SEQ ID NO:76, 92, 68, 84, 66, 70, 72, 74, 78, 80, 82,

[0186] 86, 88, 90, 94 or 96.

[0187] In some embodiments, the TCR comprises a TCRp chain variable domain encoded by a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 77, 93, 69, 85, 67, 71 , 73, 75, 79, 81 , 83,

[0188] 87, 89, 91 , 95 or 97.

[0189] In some embodiments, the TCR comprises a TCRa chain variable domain encoded by a nucleotide sequence having at least 80% sequence identity to SEQ ID NO:76, 92, 68, 84, 66, 70, 72, 74, 78, 80, 82,

[0190] 86, 88, 90, 94 or 96; in combination with a TCRp chain variable domain encoded by a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 77, 93, 69, 85, 67, 71 , 73, 75, 79, 81 , 83,

[0191] 87, 89, 91 , 95 or 97.

[0192] In some embodiments, the TCR comprises:

[0193] (a)

[0194] (i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:76 in combination with

[0195] (ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:77;

[0196] (b)

[0197] (i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:92 in combination with

[0198] (ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:93;

[0199] (c)

[0200] (i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:68 in combination with

[0201] (ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:69;

[0202] (d)

[0203] (i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:84 in combination with

[0204] (ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:85;

[0205] (e)

[0206] (i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:66 in combination with

[0207] (ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:67;

[0208] (f)

[0209] (i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NQ:70 in combination with

[0210] (ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:71 ;

[0211] (g) (i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:72 in combination with

[0212] (ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:73;

[0213] (h)

[0214] (i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:74 in combination with

[0215] (ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:75;

[0216] (i)

[0217] (i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:78 in combination with

[0218] (ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:79;

[0219] 0)

[0220] (i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NQ:80 in combination with

[0221] (ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:81 ;

[0222] (k)

[0223] (i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:86 in combination with

[0224] (ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:87;

[0225] (l)

[0226] (i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:88 in combination with

[0227] (ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:89;

[0228] (m)

[0229] (i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NQ:90 in combination with

[0230] (ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:91 ;

[0231] (n)

[0232] (i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:94 in combination with

[0233] (ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:95;

[0234] (o)

[0235] (i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:96 in combination with

[0236] (ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:97.

[0237] In some embodiments, the TCR comprises a TCRa chain constant domain comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:162 or 165.

[0238] In some embodiments, the TCR comprises a TCRp chain constant domain comprising an amino acid sequence having at least 80% sequence identity to an amino acid sequence selected from SEQ ID NO:163, 164 or 166. Also provided is a multispecific antigen-binding molecule comprising a TCR or an antigen-binding fragment of a TCR according to the present disclosure.

[0239] In some embodiments, the multispecific antigen-binding molecule further comprises an antigen-binding moiety capable of binding to a molecule expressed by an immune cell, optionally wherein the molecule expressed by an immune cell is CD3.

[0240] Also provided is a nucleic acid comprising a nucleotide sequence encoding all or part of a TCR according to the present disclosure.

[0241] Also provided is an isolated nucleic acid comprising a nucleotide sequence having at least 80% sequence identity to:

[0242] (a) SEQ ID NO:76, 77, 92 and / or 93;

[0243] (b) SEQ ID NO:68, 69, 84 and / or 85;

[0244] (c) SEQ ID NO:66, 67, 82 and / or 83;

[0245] (d) SEQ ID NQ:70, 71 , 86 and / or 87;

[0246] (e) SEQ ID NO:72, 73, 88 and / or 89;

[0247] (f) SEQ ID NO:74, 75, 90 and / or 91 ;

[0248] (g) SEQ ID NO:78, 79, 94 and / or 95; or

[0249] (h) SEQ ID NQ:80, 81 , 96 and / or 97.

[0250] Also provided is an isolated nucleic acid comprising a nucleotide sequence having at least 80% sequence identity to:

[0251] (a) SEQ ID NO:92 and / or 93;

[0252] (b) SEQ ID NO:84 and / or 85;

[0253] (c) SEQ ID NO:82 and / or 83;

[0254] (d) SEQ ID NO:86 and / or 87;

[0255] (e) SEQ ID NO:88 and / or 89;

[0256] (f) SEQ ID NQ:90 and / or 91 ;

[0257] (g) SEQ ID NO:94 and / or 95; or

[0258] (h) SEQ ID NO:96 and / or 97.

[0259] Also provided is a nucleic acid comprising a nucleotide sequence encoding all or part of a multispecific antigen-binding molecule according to the present disclosure.

[0260] Also provided is a vector comprising a nucleic acid or nucleotide sequence according to the present disclosure.

[0261] In some embodiments, the vector further comprises a nucleotide sequence of SEQ ID NO:171 . Also provided is a host cell comprising and / or expressing: a TCR, a multispecific antigen-binding molecule, a nucleic acid, and / or a vector according to the present disclosure.

[0262] In some embodiments, the host cell is an immune cell.

[0263] In some embodiments, the host cell is a T lymphocyte.

[0264] Also provided is a composition comprising a TCR, a multispecific antigen-binding molecule, a nucleic acid, a vector, or a host cell according to the present disclosure.

[0265] In some embodiments, the composition further comprises a pharmaceutical carrier or excipient.

[0266] Also provided is a TCR, a multispecific antigen-binding molecule, a nucleic acid, a vector, a host cell, or a composition according to the present disclosure, for use as a medicament.

[0267] Also provided is a method of treating a disease or condition associated with expression of WT-1 , comprising administering a TCR, a multispecific antigen-binding molecule, a nucleic acid, a vector, a host cell, or a composition according to the present disclosure to a subject.

[0268] Also provided is a TCR, a multispecific antigen-binding molecule, a nucleic acid, a vector, a host cell, or a composition according to the present disclosure, for use in a method of treating a disease or condition associated with expression of WT-1.

[0269] Also provided is a TCR, a multispecific antigen-binding molecule, a nucleic acid, a vector, a host cell, or a composition according to the present disclosure, in the manufacture of a medicament for the treatment of a disease or condition associated with expression of WT-1 .

[0270] In some embodiments, the disease or condition is cancer or a neoplasm.

[0271] In some embodiments, the disease or condition is selected from: a WT-1 -associated cancer, a cancer comprising cells comprising / presenting a peptide of a WT-1 antigen, a cancer comprising cells comprising / presenting a peptide of SEQ ID NO:1 , 179, 180, 181 or 182, a cancer comprising cells comprising / presenting a peptide of WT-1 and a major histocompatibility complex (MHC) molecule comprising an MHC class I molecule encoded by a HLA-A*02:01 allele, a cancer comprising cells comprising / presenting a peptide of SEQ ID NO:1 , 179, 180, 181 or 182, and a major histocompatibility complex (MHC) molecule comprising an MHC class I molecule encoded by a HLA-A*02:01 allele, a cancer comprising cells comprising / presenting a peptide of WT-1 and a major histocompatibility complex (MHC) molecule comprising an MHC class I molecule encoded by a HLA-A*02:07 allele, a cancer comprising cells comprising / presenting a peptide of SEQ ID NO:1 , 179, 180, 181 or 182, and a major histocompatibility complex (MHC) molecule comprising an MHC class I molecule encoded by a HLA- A*02:07 allele, a cancer comprising cells comprising / presenting a peptide of WT-1 and a major histocompatibility complex (MHC) molecule comprising an MHC class I molecule encoded by a HLA- A*02:02 allele, a cancer comprising cells comprising / presenting a peptide of SEQ ID NO:1 , 179, 180, 181 or 182, and a major histocompatibility complex (MHC) molecule comprising an MHC class I molecule encoded by a HLA-A*02:02 allele.

[0272] In some embodiments, the disease or condition is selected from: a solid cancer, a hematological cancer, gastric cancer (e.g., gastric carcinoma, gastric adenocarcinoma, gastrointestinal cancer, gastrointestinal adenocarcinoma), liver cancer (hepatocellular carcinoma, cholangiocarcinoma), head and neck cancer (e.g., head and neck squamous cell carcinoma), oral cavity cancer (e.g., oropharyngeal cancer (e.g., oropharyngeal carcinoma), oral cancer, oral squamous cell carcinoma (OSCC), laryngeal cancer, nasopharyngeal carcinoma, oesophageal cancer), colorectal cancer (e.g., colorectal carcinoma), colon cancer, colon carcinoma, cervical carcinoma, prostate cancer, lung cancer (e.g., NSCLC, small cell lung cancer, lung adenocarcinoma, squamous lung cell carcinoma), bladder cancer, urothelial carcinoma, urogenital cancer, skin cancer (e.g., melanoma, advanced melanoma), renal cell cancer (e.g., renal cell carcinoma), ovarian cancer (e.g., ovarian carcinoma), mesothelioma, breast cancer (e.g., triple-negative, Luminal A, Luminal B, and / or HER2+ breast cancer), brain cancer (e.g., glioblastoma), prostate cancer, pancreatic cancer, endometrial cancer, a myeloid hematologic malignancy, a lymphoblastic hematologic malignancy, myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute lymphoblastic leukemia (ALL), lymphoma, non-Hodgkin’s lymphoma (NHL), thymoma and multiple myeloma (MM).

[0273] In some embodiments, the treatment further comprises administering one or more immune modulating agents to the subject, wherein the one or more immune modulating agents are selected from the group consisting of: immune cell-depleting agents, cytokines, TLR agonists, RIG-I like receptor (RLR) agonists, immune checkpoint inhibitors, chemotherapeutic agents, antibodies, radiotherapy, and a combination thereof.

[0274] In some embodiments, the treatment comprises:

[0275] (a) modifying a cell to express or comprise the TCR or a nucleic acid or vector encoding the TCR, and / or

[0276] (b) administering a / the cell modified to express or comprise a TCR or nucleic acid or vector encoding the TCR to a subject.

[0277] In some embodiments, the treatment comprises administering at least one TCR, multispecific antigenbinding molecule, nucleic acid, vector, cell and / or composition according to the present disclosure to a subject, in combination with at least one non-identical TCR, multispecific antigen-binding molecule, nucleic acid, vector, cell and / or composition.

[0278] In some embodiments, the subject comprises a cell that expresses WT-1 , and / or that expresses a WT-1 peptide of SEQ ID NO:1 , 179, 180, 181 or 182. Description

[0279] The present invention relates to TCR sequences for use in TCR-based immunotherapies against cells expressing WT-1 .

[0280] T cells are the most actively studied cell type in the growing field of adoptive cellular therapeutics. T cells interact specifically with the target of their T cell receptor (TCR), enabling highly specific responses with minimal side effects. These potentially highly effective and specific responses can be engineered towards novel antigens and targets by inserting a new receptor with the desired specificity into a T cell. However, development of entirely new types of receptors is time consuming, expensive, and fails to take advantage of the fact that, through development of the endogenous T cell repertoire, the body naturally produces TCRs that bind almost any possible antigenic target. The ability to obtain human T cells and replace their endogenous TCR with a TCR having a desired antigen specificity could be transformative in the development and application of adoptive T cell therapies.

[0281] In various embodiments, the present invention provides human T cell receptors (TCRs) that are capable of binding to WT-1 antigenic peptides which are associated with diseases such as cancer. Also provided is the use of the TCRs of the invention in fusion constructs, nucleic acids encoding the TCRs of the invention, vectors comprising said nucleic acids, and host cells comprising or expressing TCRs of the invention. The present invention also provides TCRs of the invention for use as a medicament and / or in treating a disease or condition associated with expression of WT-1 , such as cancer.

[0282] Effector cells transduced with the presently claimed TCRs show dose-dependent T cell activation and dose-dependent cytotoxic activity. Primary T cells transduced with TCRs of the present invention demonstrate cell killing activity against WT-1 -expressing cell lines, such chronic myelogenous leukemia (CML) cell line K562, ovarian cancer cell line COV434, glioblastoma cell line SF539, and pancreatic cancer cell line Panel . Accordingly, the T cells transduced with TCRs of the present invention exhibit cell killing activity towards cancer cell lines displaying the WT-1 antigen of interest on the corresponding HLA molecule.

[0283] WT-1

[0284] WT-1 (Wilms’ Tumor Antigen 1) is the protein identified by UniProtKB: P19544. The canonical isoform of human WT-1 has the amino acid sequence of P19544-1 (v2, 1991-08-01 ; SEQ ID NO:169). Alternative initiation sites and alternative splicing of mRNA encoded by the human WT1 gene yields eight main isoforms: isoform 1 (SEQ ID NO:169), isoform 2 (SEQ ID NO:184), isoform 3 (SEQ ID NO:185), isoform 4 (SEQ ID NO:186), isoform 6 (SEQ ID NO:187), isoform 7 (SEQ ID NO:188), isoform 8 (SEQ ID NO:189), isoform 9 (SEQ ID NQ:190). Isoform 1 has the amino acid sequence of SEQ ID NO:169. Isoform 2 differs from isoform 1 in that positions 250-266 and 408-410 are absent. Isoform 3 differs from isoform 1 in that positions 250-266 are missing. Isoform 4 differs from isoform 1 in that positions 408 to 410 are missing. Isoform 6 differs from isoform 1 in that positions 1 to 144 and 408 to 410 are absent, and positions 145 to 147 are replaced with a different sequence of amino acids. Isoform 7 differs from isoform 1 in that it has an extended N-terminus with 73 additional amino acids. Isoform 8 differs from isoform 1 in that positions 250 to 266 are absent and in that it has an extended N-terminus with 73 additional amino acids. Isoform 9 differs from isoform 1 in that positions 1 to 144 and 250 to 266 are absent, and in that positions 145 to 147 are replaced with a different sequence of amino acids.

[0285] WT-1 is a zinc finger transcription factor whose expression in normal adult tissue is rare but is overexpressed in leukemias of multiple lineages and a wide range of solid tumors. WT-1 comprises four zinc finger domains at the C-terminus and a glutamine and proline-rich domain at the N-terminus. The WT1 proteins seem to perform two main functions. They regulate the transcription of a variety of target genes and may be involved in post-transcriptional processing of RNA. WT-1 plays an important role in cell proliferation, differentiation, apoptosis, organ development and the maintenance of several adult tissues. In disease, WT-1 is thought to inhibit cell apoptosis by transcriptional activation and / or upregulation of proto-oncogenes. It is also thought to promote invasion, migration and metastasis, facilitate angiogenesis and confer drug resistance to cancer cells. In cancers WT1 expression may be a biomarker and a prognostic indicator.

[0286] The structure and function of WT-1 is described in e.g., Scharnhorst et al., Gene (2001) 273:141-161 , which is hereby incorporated by reference in its entirety.

[0287] Reference herein to ‘WT-1 ’ generally refers to the canonical isoform of the human WT-1 ( / .e., isoform 1), but also contemplates isoforms, fragments, variants (including mutants) and homologues thereof ( / .e., from other species, e.g., non-human mammalian species {e.g., a non-human primate, e.g., rhesus, cynomolgous; e.g., a rodent, e.g., rat or mouse).

[0288] As used herein, a ‘fragment’, ‘variant’ or ‘homologue’ of a protein may optionally be characterised as having at least 60% amino acid sequence identity, e.g., one of >70%, >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98% or >99% amino acid sequence identity to the amino acid sequence of the reference protein {e.g., the canonical isoform of the human protein). In some embodiments fragments / variants / isoforms / homologues may be characterised by ability to perform a function performed by the reference protein.

[0289] A ‘fragment’ generally refers to a fraction of the reference protein. A ‘variant’ generally refers to a protein having an amino acid sequence comprising one or more amino acid substitutions, insertions, deletions or other modifications relative to the amino acid sequence of the reference protein, but retaining a considerable degree of sequence identity {e.g., at least 60%) to the amino acid sequence of the reference protein. An ‘isoform’ generally refers to a variant of the reference protein expressed by the same species as the species of the reference protein {e.g., human WT-1 isoforms 1 to 4 and 6 to 9 are all isoforms of one another). A ‘homologue’ generally refers to a variant of the reference protein produced by a different species as compared to the species of the reference protein. For example, human WT-1 isoform 1 (P19544-1 v2; SEQ ID NO:169) and mouse WT-1 (UniProt: P22561-1 , v2 1991-08-01) are homologues of one another. Homologues include orthologues.

[0290] A ‘fragment’ may be of any length (by number of amino acids), although may optionally be at least 20% of the length of the reference protein (that is, the protein from which the fragment is derived) and may have a maximum length of one of 50%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the length of the reference protein. A fragment of WT-1 may have a minimum length of one of 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 amino acids, and may have a maximum length of one of 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 amino acids.

[0291] Isoforms, fragments, variants or homologues may optionally be functional isoforms, fragments, variants or homologues, e.g., having a functional property / activity of the reference WT-1 {e.g., human WT-1 isoform 1), as determined by analysis by a suitable assay for the functional property / activity. For example, an isoform, fragment, variant or homologue of WT-1 may display association with an interaction partner of WT-1.

[0292] In some embodiments, the WT-1 is WT-1 from a mammal {e.g., a primate (rhesus, cynomolgous, nonhuman primate or human) and / or a rodent {e.g., rat or murine) WT-1). Isoforms, fragments, variants or homologues of WT-1 may optionally be characterized as having at least 60% amino acid sequence identity, e.g., one of >70%, >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity to the amino acid sequence of a WT-1 isoform from a given species, e.g., human.

[0293] In some embodiments, the WT-1 comprises, or consists of, an amino acid sequence having at least 60% amino acid sequence identity, e.g., one of >70%, >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity to one of SEQ ID NOs:169, 184, 185, 186, 187, 188, 189 or 190. In some embodiments, the WT-1 comprises, or consists of, an amino acid sequence having at least 60% amino acid sequence identity, e.g., one of >70%, >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity to one of SEQ ID NOs:169, 184, 185, 186, 188, or 189.

[0294] In some embodiments, a fragment of WT-1 comprises, or consists of, an amino acid sequence having at least 60% amino acid sequence identity, e.g., one of >70%, >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity to SEQ ID NO:1 , 179, 180, 181 or 182. In some embodiments, a fragment of WT-1 comprises, or consists of, an amino acid sequence having at least 60% amino acid sequence identity, e.g., one of >70%, >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity to SEQ ID NO:1.

[0295] T Cell Receptors

[0296] The present disclosure provides antigen-binding molecules and fragments, chains, polypeptides and complexes thereof, e.g., T cell receptors.

[0297] The term ‘T cell receptor’ (TCR), as used herein, refers to a heteromeric cell-surface receptor capable of specifically interacting with a target antigen. Herein, a ‘TCR’ or an antigen-binding fragment thereof may also be referred to as an ‘antigen-binding molecule’. As used herein, ‘TCR’ includes but is not limited to: naturally occurring and non-naturally occurring TCRs; full-length TCRs and antigen-binding portions thereof, chimeric TCRs; TCR fusion constructs; and synthetic TCRs. In humans, TCRs are expressed on the surface of T cells, and they are responsible for T cell recognition and targeting of antigen presenting cells. Antigen presenting cells (APC) display fragments of foreign or self-proteins (antigens) complexed with the major histocompatibility complex (MHC; also referred to herein as complexed with a HLA molecule, e.g., a HLA class I or class II molecule). A TCR recognizes and binds to the antigemHLA complex and recruits CD3 (expressed by T cells), activating the TCR. The activated TCR initiates downstream signaling and an immune response, including the destruction of the APC.

[0298] In general, a TCR can comprise two chains, an alpha chain and a beta chain (or less commonly a gamma chain and a delta chain), interconnected by disulfide bonds to form a heterodimeric receptor. Each chain comprises a variable domain (e.g., alpha chain variable domain or beta chain variable domain) and a constant region (e.g., alpha chain constant region or beta chain constant region). The variable domain is located distal to the cell membrane, and interacts with an antigen. A variable domain may also be referred to herein as a ‘variable region’. The constant region is located proximal to the cell membrane, acts to anchor the protein in the cell membrane, and associates with invariant subunits of the CD3 signaling apparatus. A TCR can further comprise a transmembrane region and a short cytoplasmic tail. As used herein, the term ‘constant region’ encompasses the transmembrane region and the cytoplasmic tail, when present, as well as the traditional ‘constant region.’

[0299] The variable domains can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FR). Each alpha chain variable domain and beta chain variable domain comprises three CDRs and four FRs: FR1 , CDR1 , FR2, CDR2, FR3, CDR3, FR4.

[0300] Each variable domain contains a binding domain that interacts with an antigen. This may be called an antigen-binding domain or antigen-binding fragment of a TCR. Together, the 6 CDRs form the majority of the antigen binding site of the TCR, thus conferring onto each TCR its specificity (Schroeder and Cavacini, J Allergy Clin Immunol 125(202):S41-S52 (2010); Bhati et al., Protein Science 23:260-272 (2014), which are hereby incorporated by reference in their entirety). The CDRs interact with a complex between an antigenic peptide bound to a protein encoded by the major histocompatibility complex (pepMHC) (Davis and Bjorkman (1988) Nature, 334, 395-402; Davis et al. (1998) Annu Rev Immunol, 16, 523-544; Murphy (2012), xix, 868 p., which are hereby incorporated by reference in their entirety). Although all three CDRs on each chain are involved in antigen binding, CDR3 is believed to be the primary antigen binding region, interacting with the peptide in the HLA complex groove during TCR binding. CDR1 and CDR2 are believed to primarily recognize the HLA complex.

[0301] The term CDR3 used herein describes the CDR3 region optionally including the fixed C’-terminal amino acid C (cysteine) and N’-terminal amino acid F (phenylalanine) or W (Tryptophan), or the respective nucleotide sequences coding for these amino acids. The CDR3 including C’-terminal C and N’-terminal F / W, or the respective codons, is also termed ‘Junction’ in the field.

[0302] Thus, provided herein are novel antigen-binding molecules, e.g., TCRs, TCR polypeptides, and fragments / chains thereof, comprising one or more of: the TCRa and TCRp CDR sequences listed in Table A, the FR sequences listed in Table B, the TCRa and TCRp variable domain amino acid sequences listed in Table C, and / or the TCRa and TCRp variable domains encoded by the nucleotide sequences listed in Table D. Where not expressly stated, and unless the context indicates otherwise, the term ‘TCR’ also includes an antigen-binding fragment or an antigen-binding portion of any TCR disclosed herein, and includes a monovalent and a divalent fragment or portion, and a single chain TCR or TCR fragment. An ‘antigenbinding molecule,’ ‘portion of a TCR,’ or ‘TCR fragment’ may refer to a portion of an TCR less than the whole. An ‘antigen-binding molecule’, ‘portion of a TCR,’ or ‘TCR fragment’ can include the antigenic complementarity determining regions (CDRs).

[0303] The term ‘TCR’ is not limited to naturally occurring TCRs bound to the surface of a T cell. As used herein, the term ‘TCR’ further refers to a TCR described herein that is expressed on the surface of a cell other than a T cell (e.g., a cell that naturally expresses or that is modified to express CD3, as described herein), or a TCR described herein that is free from a cell membrane (e.g., an isolated TCR or a soluble TCR).

[0304] Thus, in some embodiments, an antigen-binding molecule, e.g., TCR or TCR chain / polypeptide, described herein is provided in isolated or purified form. For example, articles according to the present disclosure may be isolated / purified from naturally-occurring biological material. In some embodiments, an antigen-binding molecule described herein, e.g., TCR or TCR chain / polypeptide, is soluble. In some embodiments, an antigen-binding molecule described herein, e.g., TCR or TCR chain / polypeptide, (or a nucleotide sequence encoding the antigen-binding molecule) is comprised within and / or expressed by a cell, e.g., a cell as described herein. In some embodiments, the TCR chain sequences are listed together with the leader sequence preceding FR1. The leader sequence regulates gene expression of the protein but is not present in the mature TCR protein. Example leader sequences for TCRa and TCRp chains are provided in SEQ ID NO 170.

[0305] The present invention provides an antigen-binding molecule, e.g., a TCR or fragment / chain thereof, that binds to a WT-1 -derived antigenic peptide, such as when presented by a major histocompatibility complex (MHC) molecule.

[0306] In some embodiments, the antigen-binding molecule comprises the CDRs of a TCR that binds to a WT-1- derived peptide (e.g., such as when presented by a major histocompatibility complex (MHC) molecule). In some embodiments, the antigen-binding molecule comprises the FRs of a TCR that binds to a WT-1- derived peptide (e.g., such as when presented by a major histocompatibility complex (MHC) molecule). In some embodiments, the antigen-binding molecule comprises the CDRs and the FRs of a TCR that binds to a WT-1 -derived peptide (e.g., such as when presented by a major histocompatibility complex (MHC) molecule). That is, in some embodiments, the antigen-binding molecule comprises the TCRa variable domain and the TCRp variable domain of a TCR that binds to a WT-1 -derived peptide (e.g., such as when presented by a major histocompatibility complex (MHC) molecule). The WT-1 -derived peptide may be SEQ ID NO:1 , 179, 180, 181 or 182. The WT-1 -derived peptide may be SEQ ID NO:1 , 179, 180, 181 or 182 and may be presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide, e.g., encoded by HLA-A*02, e.g., encoded by HLA-A*02:01.

[0307] In some embodiments, the antigen-binding molecule comprises the CDRs, FRs, and / or the alpha and / or beta chain variable domains of a TCR as described herein, or CDRs, FRs, and / or the alpha and / or beta chain variable domains which are derived from those of a TCR described herein. In some embodiments, a TCR is selected from TCR_A0431 , TCR_A0427, TCR_A0426, TCR_A0428, TCR_A0429, TCR_A0430, TCR_A0432, and TCR_A0464. In some embodiments, a TCR is selected from TCR_A0431 and TCR_A0427.

[0308] In some embodiments, an antigen-binding molecule is selected from TCR_A0431 , TCR_A0427, TCR_A0426, TCR_A0428, TCR_A0429, TCR_A0430, TCR_A0432, and TCR_A0464. In some embodiments, an antigen-binding molecule is selected from TCR_A0431 and TCR_A0427.

[0309] In some embodiments the antigen-binding molecule comprises a TCRa chain, or a fragment thereof. In some embodiments, the TCRa chain comprises a TCRa variable domain.

[0310] In some embodiments, the TCRa variable domain, comprises a CDR3a having an amino acid sequence of SEQ ID NO:34, 10, 4, 16, 22, 28, 40 or 46, or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:34, 10, 4, 16, 22, 28, 40 or 46 are substituted with another amino acid. In some embodiments, the TCRa variable domain, comprises a CDR3a having an amino acid sequence of SEQ ID NO:34, or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:34 are substituted with another amino acid. In some embodiments, the TCRa chain, e.g., TCRa chain variable domain, comprises a CDR3a having an amino acid sequence of SEQ ID NQ:10, or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NQ:10.

[0311] In some embodiments the antigen-binding molecule comprises a TCRp chain, or a fragment thereof. In some embodiments, the TCRp chain comprises a TCRp variable domain.

[0312] In some embodiments, the TCRp variable domain, comprises a CDR3p having an amino acid sequence of SEQ ID NO:37, 13, 7, 19, 25, 31 , 43 or 49, or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 37, 13, 7, 19, 25, 31 , 43 or 49 are substituted with another amino acid. In some embodiments, the CDR3p has an amino acid sequence of SEQ ID NO:37, or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:37 are substituted with another amino acid. In some embodiments, the CDR3p has an amino acid sequence of SEQ ID NO:13, or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 37 are substituted with another amino acid.

[0313] In some embodiments, the antigen-binding molecule comprises: (i) a TCRa variable domain comprising a CDR3a having an amino acid sequence of SEQ ID NO:34, 10, 4, 16, 22, 28, 40 or 46, or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:34, 10, 4, 16, 22, 28, 40 or 46 are substituted with another amino acid; in combination with (ii) a TCRp variable domain comprising a CDR3p having an amino acid sequence of SEQ ID NO:37, 13, 7, 19, 25, 31 , 43 or 49, or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 37, 13, 7, 19, 25, 31 , 43 or 49 are substituted with another amino acid.

[0314] In some embodiments, the antigen-binding molecule comprises: (i) a TCRa variable domain comprising a CDR3a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR3a are substituted with another amino acid) as indicated in Column A of row 1 , 2, 3, 4, 5, 6, 7, or 8 of Table A; in combination with (ii) a TCRp variable domain comprising a CDR3p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR3p are substituted with another amino acid) as indicated in Column B of row 1 , 2, 3, 4, 5, 6, 7, or 8 of Table A; wherein the CDR3a amino acid sequence from Column A and the CDR3p amino acid sequence from Column B are selected from the same row. By way of illustration, in some embodiments, the antigen-binding molecule comprises: (i) a TCRa variable domain comprising a CDR3a having an amino acid sequence of SEQ ID NO:34 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 34 are substituted with another amino acid) (row 6); in combination with (ii) a TCRp variable domain comprising a CDR3p having an amino acid sequence of SEQ ID NO:37 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:37 are substituted with another amino acid) (row 6).

[0315] By way of further illustration, in some embodiments, the antigen-binding molecule comprises: (i) a TCRa variable domain comprising a CDR3a having an amino acid sequence of SEQ ID NO:10 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NQ:10 are substituted with another amino acid) (row 2); in combination with (ii) a TCRp variable domain comprising a CDR3p having an amino acid sequence of SEQ ID NO:13 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:13 are substituted with another amino acid) (row 2).

[0316] In some embodiments, the antigen-binding molecule comprises a TCRa variable domain comprising: a CDR1a having an amino acid sequence of SEQ ID NO:32, 8, 2, 14, 20, 26, 38 or 44 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:32, 8, 2, 14, 20, 26, 38 or 44 are substituted with another amino acid); a CDR2a having an amino acid sequence of SEQ ID NO:33, 9, 3, 15, 21 , 27, 39 or 45 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:33, 9, 3, 15, 21 , 27, 39 or 45 are substituted with another amino acid); and / or a CDR3a having an amino acid sequence of SEQ ID NO:34, 10, 4, 16, 22, 28, 40 or 46 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 34, 10, 4, 16, 22, 28, 40 or 46 are substituted with another amino acid).

[0317] In some embodiments, the antigen-binding molecule comprises a TCRa variable domain comprising CDR1a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR1a are substituted with another amino acid), CDR2a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR2a are substituted with another amino acid) and / or CDR3a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR3a are substituted with another amino acid), as indicated in Column A of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table A, wherein the CDR1a, CDR2a and / or CDR3a sequences of Column A are selected from the same row of Table A.

[0318] By way of illustration, in some embodiments, the antigen-binding molecule comprises a TCRa variable domain comprising:

[0319] CDR1a having an amino acid sequence of SEQ ID NO:32 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:32 are substituted with another amino acid) (row 6);

[0320] CDR2a having an amino acid sequence of SEQ ID NO:33 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:33 are substituted with another amino acid) (row 6); and / or

[0321] CDR3a having an amino acid sequence of SEQ ID NO:34 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:34 are substituted with another amino acid) (row 6). By way of further illustration, in some embodiments, the antigen-binding molecule comprises a TCRa variable domain comprising:

[0322] CDR1a having an amino acid sequence of SEQ ID NO:8 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:8 are substituted with another amino acid) (row 2);

[0323] CDR2a having an amino acid sequence of SEQ ID NO:9 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:9 are substituted with another amino acid) (row 2); and / or

[0324] CDR3a having an amino acid sequence of SEQ ID NQ:10 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NQ:10 are substituted with another amino acid) (row 2).

[0325] In some embodiments, the antigen-binding molecule comprises a TCRa variable domain comprising: a FR1 having an amino acid sequence of SEQ ID NO:138, 106, 98, 114, 122, 130, 146 or 154 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 138, 106, 98, 114, 122, 130, 146 or 154 are substituted with another amino acid); a FR2 having an amino acid sequence of SEQ ID NO:139, 107, 99, 115, 123, 131 , 147 or 155 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 139, 107, 99, 115, 123, 131 , 147 or 155 are substituted with another amino acid); a FR3 having an amino acid sequence of SEQ ID NO:140, 108, 100, 116, 124, 132, 148 or 156 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 140, 108, 100, 116, 124, 132, 148 or

[0326] 156 are substituted with another amino acid); and / or, a FR4 having an amino acid sequence of SEQ ID NO:141 , 109, 101 , 117, 125, 133, 149 or 157 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 141 , 109, 101 , 117, 125, 133, 149 or

[0327] 157 are substituted with another amino acid).

[0328] In some embodiments, the antigen-binding molecule comprises a TCRa variable domain comprising FR1 (or a variant thereof in which 1 or 2 or 3 amino acids in FR1 are substituted with another amino acid), FR2 (or a variant thereof in which 1 or 2 or 3 amino acids in FR2 are substituted with another amino acid), FR3 (or a variant thereof in which 1 or 2 or 3 amino acids in FR3 are substituted with another amino acid) and / or FR4 (or a variant thereof in which 1 or 2 or 3 amino acids in FR4 are substituted with another amino acid) as indicated in Column A of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table B, wherein the FR1 , FR2, FR3 and / or FR4 sequences of Column A are selected from the same row of Table B.

[0329] By way of illustration, in some embodiments, the antigen-binding molecule comprises a TCRa variable domain comprising FR1 , FR2, FR3 and / or FR4 having an amino acid sequence of SEQ ID NO: 138, 139,

[0330] 140 and 141 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 138, 139, 140 and / or

[0331] 141 are substituted with another amino acid), respectively (row 6).

[0332] By way of further illustration, in some embodiments, the antigen-binding molecule comprises a TCRa variable domain comprising FR1 , FR2, FR3 and / or FR4 having an amino acid sequence of SEQ ID NO: 106, 107, 108 and 109 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 106, 107, 108 and / or 109 are substituted with another amino acid), respectively (row 2).

[0333] In some embodiments, the antigen-binding molecule comprises a TCRa variable domain comprising CDR1a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR1a are substituted with another amino acid), CDR2a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR2a are substituted with another amino acid) and / or CDR3a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR3a are substituted with another amino acid), as indicated in Column A of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table A, wherein the CDR1a, CDR2a and / or CDR3a sequences of Column A are selected from the same row of Table A; and FR1 (or a variant thereof in which 1 or 2 or 3 amino acids in FR1 are substituted with another amino acid), FR2 (or a variant thereof in which 1 or 2 or 3 amino acids in FR2 are substituted with another amino acid), FR3 (or a variant thereof in which 1 or 2 or 3 amino acids in FR3 are substituted with another amino acid) and / or FR4 (or a variant thereof in which 1 or 2 or 3 amino acids in FR4 are substituted with another amino acid) as indicated in Column A of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table B, wherein the FR1 , FR2, FR3 and / or FR4 sequences of Column A are selected from the same row of Table B.

[0334] In some embodiments, the antigen-binding molecule comprises a TCRa variable domain comprising CDR1a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR1a are substituted with another amino acid), CDR2a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR2a are substituted with another amino acid) and / or CDR3a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR3a are substituted with another amino acid), as indicated in Column A of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table A, wherein the CDR1a, CDR2a and / or CDR3a sequences of Column A are selected from the same row of Table A; and FR1 (or a variant thereof in which 1 or 2 or 3 amino acids in FR1 are substituted with another amino acid), FR2 (or a variant thereof in which 1 or 2 or 3 amino acids in FR2 are substituted with another amino acid), FR3 (or a variant thereof in which 1 or 2 or 3 amino acids in FR3 are substituted with another amino acid) and / or FR4 (or a variant thereof in which 1 or 2 or 3 amino acids in FR4 are substituted with another amino acid) as indicated in Column A of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table B, wherein the FR1 , FR2, FR3 and / or FR4 sequences of Column A are selected from the same row of Table B; wherein the CDR1a, CDR2a and / or CDR3a sequences of Column A of Table A and the FR1 , FR2, FR3 and / or FR4 of Column A of Table B are selected from rows having the same number.

[0335] By way of illustration, in some embodiments, the antigen-binding molecule comprises a TCRa variable domain comprising: CDR1a, CDR2a and / or CDR3a having an amino acid sequence of SEQ ID NO:32, 33 and 34 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:32, 33 and / or 34 are substituted with another amino acid), respectively (row 6); and FR1 , FR2, FR3 and / or FR4 having an amino acid sequence of SEQ ID NO: 138, 139, 140 and 141 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 138, 139, 140 and / or 141 are substituted with another amino acid), respectively (row 6).

[0336] By way of further illustration, in some embodiments, the antigen-binding molecule comprises a TCRa variable domain comprising: CDR1a, CDR2a and / or CDR3a having an amino acid sequence of SEQ ID NO:8, 9 and 10 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:8, 9 and / or 10 are substituted with another amino acid), respectively (row 2); and FR1 , FR2, FR3 and / or FR4 having an amino acid sequence of SEQ ID NQ:106, 107, 108 and 109 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NQ:106, 107, 108 and / or 109 are substituted with another amino acid), respectively (row 2). In some embodiments, the antigen-binding molecule comprises a TCRa variable domain having an amino acid sequence having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:60, 52, 50, 54, 56, 58, 62 or 64. In some embodiments, the antigen-binding molecule comprises a TCRa variable domain having an amino acid sequence having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:60. In some embodiments, the antigen-binding molecule comprises a TCRa variable domain having an amino acid sequence having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:52.

[0337] In some embodiments, the antigen-binding molecule comprises a TCRp variable domain comprising: a CDR1 p having an amino acid sequence of SEQ ID NO:35, 11 , 5, 17, 23, 29, 41 or 47 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 35, 11 , 5, 17, 23, 29, 41 or 47 are substituted with another amino acid); a CDR2p having an amino acid sequence of SEQ ID NO:36, 12, 6, 18, 24, 30, 42 or 48 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:36, 12, 6, 18, 24, 30, 42 or 48 are substituted with another amino acid); and / or a CDR3p having an amino acid sequence of SEQ ID NO:37, 13, 7, 19, 25, 31 , 43 or 49 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:37, 13, 7, 19, 25, 31 , 43 or 49 are substituted with another amino acid).

[0338] In some embodiments, the antigen-binding molecule comprises a TCRp variable domain comprising CDR1 p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR1 p are substituted with another amino acid), CDR2p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR2p are substituted with another amino acid) and / or CDR3p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR3p are substituted with another amino acid) as indicated in Column B of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table A, wherein the CDR1 p, CDR2p and / or CDR3p sequences of Column B are selected from the same row of Table A.

[0339] By way of illustration, in some embodiments, the antigen-binding molecule comprises a TCRp variable domain comprising:

[0340] CDR1 p having an amino acid sequence of SEQ ID NO:35 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:35 are substituted with another amino acid) (row 6);

[0341] CDR2p having an amino acid sequence of SEQ ID NO:36 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:36 are substituted with another amino acid) (row 6); and / or

[0342] CDR3p having an amino acid sequence of SEQ ID NO:37 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:37 are substituted with another amino acid) (row 6). By way of further illustration, in some embodiments, the antigen-binding molecule comprises a TCRp variable domain comprising:

[0343] CDR1 p having an amino acid sequence of SEQ ID NO:11 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:11 are substituted with another amino acid) (row 2);

[0344] CDR2p having an amino acid sequence of SEQ ID NO:12 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:12 are substituted with another amino acid) (row 2); and / or

[0345] CDR3p having an amino acid sequence of SEQ ID NO:13 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:13 are substituted with another amino acid) (row 2).

[0346] In some embodiments, the antigen-binding molecule comprises a TCRp variable domain comprising: a FR1 having an amino acid sequence of SEQ ID NO:142, 110, 102, 118, 126, 134, 150 or 158 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 142, 110, 102, 118, 126, 134, 150 or

[0347] 158 are substituted with another amino acid); a FR2 having an amino acid sequence of SEQ ID NO:143, 111 , 103, 119, 127, 135, 151 or 159 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 143, 111 , 103, 119, 127, 135, 151 or

[0348] 159 are substituted with another amino acid); a FR3 having an amino acid sequence of SEQ ID NO:144, 112, 104, 120, 128, 136, 152 or 160 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 144, 112, 104, 120, 128, 136, 152 or

[0349] 160 are substituted with another amino acid); and / or, a FR4 having an amino acid sequence of SEQ ID NO:145, 113, 105, 121 , 129, 137, 153 or 161 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 145, 113, 105, 121 , 129, 137, 153 or

[0350] 161 are substituted with another amino acid).

[0351] In some embodiments, the antigen-binding molecule comprises a TCRp variable domain comprising FR1 (or a variant thereof in which 1 or 2 or 3 amino acids in FR1 are substituted with another amino acid), FR2 (or a variant thereof in which 1 or 2 or 3 amino acids in FR2 are substituted with another amino acid), FR3 (or a variant thereof in which 1 or 2 or 3 amino acids in FR3 are substituted with another amino acid) and / or FR4 (or a variant thereof in which 1 or 2 or 3 amino acids in FR4 are substituted with another amino acid) as indicated in Column B of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table B, wherein the FR1 , FR2, FR3 and FR4 sequences of Column B are selected from the same row of Table B.

[0352] By way of illustration, in some embodiments, the antigen-binding molecule comprises a TCRp variable domain comprising FR1 , FR2, FR3 and / or FR4 having an amino acid sequence of SEQ ID NO:142, 143, 144, and 145 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:142, 143, 144, and / or 145 are substituted with another amino acid), respectively (row 6).

[0353] By way of further illustration, in some embodiments, the antigen-binding molecule comprises a TCRp variable domain comprising FR1 , FR2, FR3 and / or FR4 having an amino acid sequence of SEQ ID NQ:110, 111 , 112 and 113 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:110, 111 , 112 and / or 113 are substituted with another amino acid), respectively (row 2).

[0354] In some embodiments, the antigen-binding molecule comprises a TCRp variable domain comprising: CDR1 p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR1 p are substituted with another amino acid), CDR2p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR2p are substituted with another amino acid) and / or CDR3p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR3p are substituted with another amino acid) as indicated in Column B of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table A, wherein CDR1 p, CDR2p and / or CDR3p sequences of Column B are selected from the same row of Table A; and FR1 (or a variant thereof in which 1 or 2 or 3 amino acids in FR1 are substituted with another amino acid), FR2 (or a variant thereof in which 1 or 2 or 3 amino acids in FR2 are substituted with another amino acid), FR3 (or a variant thereof in which 1 or 2 or 3 amino acids in FR3 are substituted with another amino acid) and / or FR4 (or a variant thereof in which 1 or 2 or 3 amino acids in FR4 are substituted with another amino acid) as indicated in Column B of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table B, wherein FR1 , FR2, FR3 and / or FR4 sequences of Column B are selected from the same row of Table B.

[0355] In some embodiments, the antigen-binding molecule comprises a TCRp variable domain comprising: CDR1 p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR1 p are substituted with another amino acid), CDR2p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR2p are substituted with another amino acid) and / or CDR3p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR3p are substituted with another amino acid) as indicated in Column B of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table A, wherein CDR1 p, CDR2p and / or CDR3p sequences of Column B are selected from the same row of Table A; and FR1 (or a variant thereof in which 1 or 2 or 3 amino acids in FR1 are substituted with another amino acid), FR2 (or a variant thereof in which 1 or 2 or 3 amino acids in FR2 are substituted with another amino acid), FR3 (or a variant thereof in which 1 or 2 or 3 amino acids in FR3 are substituted with another amino acid) and / or FR4 (or a variant thereof in which 1 or 2 or 3 amino acids in FR4 are substituted with another amino acid) as indicated in Column B of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table B, wherein FR1 , FR2, FR3 and / or FR4 sequences of Column B are selected from the same row of Table B; wherein the CDR1 p, CDR2p and / or CDR3p sequences of Column B of Table A and the FR1 , FR2, FR3 and / or FR4 of Column B of Table B are selected from rows having the same number.

[0356] By way of illustration, in some embodiments, the antigen-binding molecule comprises a TCRp variable domain comprising: CDR1 p, CDR2p and / or CDR3p having an amino acid sequence of SEQ ID NO:35, 36 and 37 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:35, 36 and / or 37 are substituted with another amino acid), respectively (row 6); and FR1 , FR2, FR3 and / or FR4 having an amino acid sequence of SEQ ID NO:142, 143, 144 and 145 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:142, 143, 144 and / or 145 are substituted with another amino acid), respectively (row 6).

[0357] By way of further illustration, in some embodiments, the antigen-binding molecule comprises a TCRp variable domain comprising: CDR1 p, CDR2p and / or CDR3p having an amino acid sequence of SEQ ID NO:11 , 12 and 13 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:11 , 12 and / or 13 are substituted with another amino acid), respectively (row 2); and FR1 , FR2, FR3 and / or FR4 having an amino acid sequence of SEQ ID NO:110, 111 , 112 and 113 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:110, 111 , 112 and / or 113 are substituted with another amino acid), respectively (row 2). In some embodiments, the antigen-binding molecule comprises a TCRp variable domain having an amino acid sequence having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:61 , 53, 51 , 55, 57, 59, 63 or 65. In some embodiments, the antigen-binding molecule comprises a TCRp variable domain having an amino acid sequence having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:61. In some embodiments, the antigen-binding molecule comprises a TCRp variable domain having an amino acid sequence having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:53.

[0358] In some embodiments, the antigen-binding molecule comprises:

[0359] (i) a TCRa variable domain comprising: a CDR1a having an amino acid sequence of SEQ ID NO:32, 8, 2, 14, 20, 26, 38 or 44 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:32, 8, 2, 14, 20, 26, 38 or 44 are substituted with another amino acid); a CDR2a having an amino acid sequence of SEQ ID NO:33, 9, 3, 15, 21 , 27, 39 or 45 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:33, 9, 3, 15, 21 , 27, 39 or 45 are substituted with another amino acid); and / or a CDR3a having an amino acid sequence of SEQ ID NO:34, 10, 4, 16, 22, 28, 40 or 46 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 34, 10, 4, 16, 22, 28, 40 or 46 are substituted with another amino acid); in combination with,

[0360] (ii) a TCRp variable domain comprising: a CDR1 p having an amino acid sequence of SEQ ID NO:35, 11 , 5, 17, 23, 29, 41 or 47 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 35, 11 , 5, 17, 23, 29, 41 or 47 are substituted with another amino acid); a CDR2p having an amino acid sequence of SEQ ID NO:36, 12, 6, 18, 24, 30, 42 or 48 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:36, 12, 6, 18, 24, 30, 42 or 48 are substituted with another amino acid); and / or a CDR3p having an amino acid sequence of SEQ ID NO:37, 13, 7, 19, 25, 31 , 43 or 49 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:37, 13, 7, 19, 25, 31 , 43 or 49 are substituted with another amino acid).

[0361] In some embodiments, the antigen-binding molecule comprises:

[0362] (i) a TCRa variable domain comprising CDR1a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR1a are substituted with another amino acid), CDR2a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR2a are substituted with another amino acid) and / or CDR3a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR3a are substituted with another amino acid), as indicated in Column A of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table A, wherein the CDR1a, CDR2a and / or CDR3a sequences of Column A are selected from the same row of Table A; in combination with

[0363] (ii) a TCRp variable domain comprising CDR1 p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR1 p are substituted with another amino acid), CDR2p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR2p are substituted with another amino acid) and / or CDR3p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR3p are substituted with another amino acid) as indicated in Column B of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table A, wherein the CDR1 p, CDR2p and / or CDR3p sequences of Column B are selected from the same row of Table A.

[0364] In some embodiments, the antigen-binding molecule comprises:

[0365] (i) a TCRa variable domain comprising CDR1a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR1a are substituted with another amino acid), CDR2a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR2a are substituted with another amino acid) and / or CDR3a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR3a are substituted with another amino acid), as indicated in Column A of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table A, wherein the CDR1a, CDR2a and / or CDR3a sequences of Column A are selected from the same row of Table A; in combination with

[0366] (ii) a TCRp variable domain comprising CDR1 p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR1 p are substituted with another amino acid), CDR2p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR2p are substituted with another amino acid) and / or CDR3p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR3p are substituted with another amino acid) as indicated in Column B of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table A, wherein the CDR1 p, CDR2p and / or CDR3p sequences of Column B are selected from the same row of Table A; wherein the CDR1a, CDR2a and / or CDR3a amino acid sequences from Column A and the CDR1 p, CDR2p and / or CDR3p amino acid sequences from Column B are selected from the same row.

[0367] By way of illustration, in some embodiments, the antigen-binding molecule comprises:

[0368] (i) a TCRa variable domain comprising: CDR1a having an amino acid sequence of SEQ ID NO:32 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:32 are substituted with another amino acid) (row 6); CDR2a having an amino acid sequence of SEQ ID NO:33 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:33 are substituted with another amino acid) (row 6); and / or CDR3a having an amino acid sequence of SEQ ID NO:34 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:34 are substituted with another amino acid) (row 6); in combination with

[0369] (ii) a TCRp variable domain comprising: CDR1 p having an amino acid sequence of SEQ ID NO:35 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:35 are substituted with another amino acid) (row 6); CDR2p having an amino acid sequence of SEQ ID NO:36 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:36 are substituted with another amino acid) (row 6); and / or CDR3p having an amino acid sequence of SEQ ID NO:37 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:37 are substituted with another amino acid) (row 6).

[0370] By way of further illustration, in some embodiments, the antigen-binding molecule comprises:

[0371] (i) a TCRa variable domain comprising: CDR1a having an amino acid sequence of SEQ ID NO:8 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:8 are substituted with another amino acid) (row 2); CDR2a having an amino acid sequence of SEQ ID NO:9 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:9 are substituted with another amino acid) (row 2); and / or CDR3a having an amino acid sequence of SEQ ID NQ:10 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NQ:10 are substituted with another amino acid) (row 2); in combination with (ii) a TCRp variable domain comprising: CDR1 p having an amino acid sequence of SEQ ID NO:11 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:11 are substituted with another amino acid) (row 2); CDR2p having an amino acid sequence of SEQ ID NO:12 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:12 are substituted with another amino acid) (row 2); and / or CDR3p having an amino acid sequence of SEQ ID NO:13 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:13 are substituted with another amino acid) (row 2).

[0372] In some embodiments, the antigen-binding molecule comprises:

[0373] (i) a TCRa variable domain comprising: a FR1 having an amino acid sequence of SEQ ID

[0374] NO: 138, 106, 98, 1 14, 122, 130, 146 or 154 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:138, 106, 98, 114, 122, 130, 146 or 154 are substituted with another amino acid); a FR2 having an amino acid sequence of SEQ ID NO:139, 107, 99, 115, 123, 131 , 147 or 155 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:139, 107, 99, 1 15, 123, 131 , 147 or 155 are substituted with another amino acid); a FR3 having an amino acid sequence of SEQ ID NQ:140, 108, 100, 116, 124, 132, 148 or 156 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 140, 108, 100, 116, 124, 132, 148 or 156 are substituted with another amino acid); and / or, a FR4 having an amino acid sequence of SEQ ID NO: 141 , 109, 101 , 117, 125, 133, 149 or 157 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 141 , 109, 101 , 117, 125, 133, 149 or 157 are substituted with another amino acid); in combination with,

[0375] (ii) a TCRp variable domain comprising: a FR1 having an amino acid sequence of SEQ ID

[0376] NO: 142, 110, 102, 118, 126, 134, 150 or 158 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:142, 1 10, 102, 118, 126, 134, 150 or 158 are substituted with another amino acid); a FR2 having an amino acid sequence of SEQ ID NO: 143, 111 , 103, 119, 127, 135, 151 or 159 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:143, 111 , 103, 119, 127, 135, 151 or 159 are substituted with another amino acid); a FR3 having an amino acid sequence of SEQ ID NO:144, 1 12, 104, 120, 128, 136, 152 or 160 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 144, 112, 104, 120, 128, 136, 152 or 160 are substituted with another amino acid); and / or, a FR4 having an amino acid sequence of SEQ ID NO: 145, 1 13, 105, 121 , 129, 137, 153 or 161 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:145, 113, 105, 121 , 129, 137, 153 or 161 are substituted with another amino acid).

[0377] In some embodiments, the antigen-binding molecule comprises:

[0378] (i) a TCRa variable domain comprising FR1 (or a variant thereof in which 1 or 2 or 3 amino acids in FR1 are substituted with another amino acid), FR2 (or a variant thereof in which 1 or 2 or 3 amino acids in FR2 are substituted with another amino acid), FR3 (or a variant thereof in which 1 or 2 or 3 amino acids in FR3 are substituted with another amino acid) and / or FR4 (or a variant thereof in which 1 or 2 or 3 amino acids in FR4 are substituted with another amino acid) as indicated in Column A of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table B, wherein the FR1 , FR2, FR3 and / or FR4 sequences of Column A are selected from the same row of Table B; in combination with

[0379] (ii) a TCRp variable domain comprising FR1 (or a variant thereof in which 1 or 2 or 3 amino acids in FR1 are substituted with another amino acid), FR2 (or a variant thereof in which 1 or 2 or 3 amino acids in FR2 are substituted with another amino acid), FR3 (or a variant thereof in which 1 or 2 or 3 amino acids in FR3 are substituted with another amino acid) and / or FR4 (or a variant thereof in which 1 or 2 or 3 amino acids in FR4 are substituted with another amino acid) as indicated in Column B of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table B, wherein the FR1 , FR2, FR3 and FR4 sequences of Column B are selected from the same row of Table B.

[0380] In some embodiments, the antigen-binding molecule comprises:

[0381] (i) a TCRa variable domain comprising FR1 (or a variant thereof in which 1 or 2 or 3 amino acids in FR1 are substituted with another amino acid), FR2 (or a variant thereof in which 1 or 2 or 3 amino acids in FR2 are substituted with another amino acid), FR3 (or a variant thereof in which 1 or 2 or 3 amino acids in FR3 are substituted with another amino acid) and / or FR4 (or a variant thereof in which 1 or 2 or 3 amino acids in FR4 are substituted with another amino acid) as indicated in Column A of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table B, wherein the FR1 , FR2, FR3 and / or FR4 sequences of Column A are selected from the same row of Table B; in combination with,

[0382] (ii) a TCRp variable domain comprising FR1 (or a variant thereof in which 1 or 2 or 3 amino acids in FR1 are substituted with another amino acid), FR2 (or a variant thereof in which 1 or 2 or 3 amino acids in FR2 are substituted with another amino acid), FR3 (or a variant thereof in which 1 or 2 or 3 amino acids in FR3 are substituted with another amino acid) and / or FR4 (or a variant thereof in which 1 or 2 or 3 amino acids in FR4 are substituted with another amino acid) as indicated in Column B of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table B, wherein the FR1 , FR2, FR3 and FR4 sequences of Column B are selected from the same row of Table B; wherein the FR1 , FR2, FR3 and / or FR4 amino acid sequences from Column A and the FR1 , FR2, FR3 and / or FR4 amino acid sequences from Column B are selected from the same row.

[0383] By way of illustration, in some embodiments, the antigen-binding molecule comprises:

[0384] (i) a TCRa variable domain comprising FR1 , FR2, FR3 and / or FR4 having an amino acid sequence of SEQ ID NO: 138, 139, 140 and 141 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 138, 139, 140 and / or 141 are substituted with another amino acid), respectively (row 6); in combination with,

[0385] (ii) a TCRp variable domain comprising FR1 , FR2, FR3 and / or FR4 having an amino acid sequence of SEQ ID NO:142, 143, 144, and 145 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:142, 143, 144, and / or 145 are substituted with another amino acid), respectively (row 6).

[0386] By way of further illustration, in some embodiments, the antigen-binding molecule comprises:

[0387] (i) a TCRa variable domain comprising FR1 , FR2, FR3 and / or FR4 having an amino acid sequence of SEQ ID NO: 106, 107, 108 and 109 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 106, 107, 108 and / or 109 are substituted with another amino acid), respectively (row 2); in combination with,

[0388] (ii) a TCRp variable domain comprising FR1 , FR2, FR3 and / or FR4 having an amino acid sequence of SEQ ID NO:110, 111 , 112 and 113 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:110, 111 , 112 and / or 113 are substituted with another amino acid), respectively (row 2).

[0389] In some embodiments, the antigen-binding molecule comprises: (i) a TCRa variable domain comprising:

[0390] CDR1a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR1a are substituted with another amino acid), CDR2a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR2a are substituted with another amino acid) and / or CDR3a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR3a are substituted with another amino acid), as indicated in Column A of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table A; and

[0391] FR1 (or a variant thereof in which 1 or 2 or 3 amino acids in FR1 are substituted with another amino acid), FR2 (or a variant thereof in which 1 or 2 or 3 amino acids in FR2 are substituted with another amino acid), FR3 (or a variant thereof in which 1 or 2 or 3 amino acids in FR3 are substituted with another amino acid) and / or FR4 (or a variant thereof in which 1 or 2 or 3 amino acids in FR4 are substituted with another amino acid) as indicated in Column A of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table B; wherein the CDR1a, CDR2a and / or CDR3a sequences of Column A of Table A, and the FR1 , FR2, FR3 and / or FR4 of Column A of Table B are selected from rows having the same number; in combination with

[0392] (ii) a TCRp variable domain comprising:

[0393] CDR1 p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR1 p are substituted with another amino acid), CDR2p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR2p are substituted with another amino acid) and / or CDR3p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR3p are substituted with another amino acid) as indicated in Column B of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table A; and

[0394] FR1 (or a variant thereof in which 1 or 2 or 3 amino acids in FR1 are substituted with another amino acid), FR2 (or a variant thereof in which 1 or 2 or 3 amino acids in FR2 are substituted with another amino acid), FR3 (or a variant thereof in which 1 or 2 or 3 amino acids in FR3 are substituted with another amino acid) and / or FR4 (or a variant thereof in which 1 or 2 or 3 amino acids in FR4 are substituted with another amino acid) as indicated in Column B of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table B; wherein the CDR1 p, CDR2p and / or CDR3p sequences of Column B of Table A, and the FR1 , FR2, FR3 and / or FR4 of Column B of Table B are selected from rows having the same number.

[0395] In some embodiments, the antigen-binding molecule comprises:

[0396] (i) a TCRa variable domain comprising:

[0397] CDR1a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR1a are substituted with another amino acid), CDR2a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR2a are substituted with another amino acid) and / or CDR3a (or a variant thereof in which 1 or 2 or 3 amino acids in CDR3a are substituted with another amino acid), as indicated in Column A of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table A; and

[0398] FR1 (or a variant thereof in which 1 or 2 or 3 amino acids in FR1 are substituted with another amino acid), FR2 (or a variant thereof in which 1 or 2 or 3 amino acids in FR2 are substituted with another amino acid), FR3 (or a variant thereof in which 1 or 2 or 3 amino acids in FR3 are substituted with another amino acid) and / or FR4 (or a variant thereof in which 1 or 2 or 3 amino acids in FR4 are substituted with another amino acid) as indicated in Column A of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table B; in combination with

[0399] (ii) a TCRp variable domain comprising: CDR1 p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR1 p are substituted with another amino acid), CDR2p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR2p are substituted with another amino acid) and / or CDR3p (or a variant thereof in which 1 or 2 or 3 amino acids in CDR3p are substituted with another amino acid) as indicated in Column B of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table A; and

[0400] FR1 (or a variant thereof in which 1 or 2 or 3 amino acids in FR1 are substituted with another amino acid), FR2 (or a variant thereof in which 1 or 2 or 3 amino acids in FR2 are substituted with another amino acid), FR3 (or a variant thereof in which 1 or 2 or 3 amino acids in FR3 are substituted with another amino acid) and / or FR4 (or a variant thereof in which 1 or 2 or 3 amino acids in FR4 are substituted with another amino acid) as indicated in Column B of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table B; wherein the CDR1a, CDR2a and / or CDR3a sequences of Column A of Table A; the FR1 , FR2, FR3 and / or FR4 of Column A of Table B; the CDR1 p, CDR2p and / or CDR3p sequences of Column B of Table A; and the FR1 , FR2, FR3 and / or FR4 of Column B of Table B are selected from rows having the same number.

[0401] By way of illustration, in some embodiments, the antigen-binding molecule comprises:

[0402] (i) a TCRa variable domain comprising: CDR1a, CDR2a and / or CDR3a having an amino acid sequence of SEQ ID NO:32, 33 and 34 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:32, 33 and / or 34 are substituted with another amino acid), respectively (row 6); and FR1 , FR2, FR3 and / or FR4 having an amino acid sequence of SEQ ID NO: 138, 139, 140 and 141 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO: 138, 139, 140 and / or 141 are substituted with another amino acid), respectively (row 6); in combination with,

[0403] (ii) a TCRp variable domain comprising: CDR1 p, CDR2p and / or CDR3p having an amino acid sequence of SEQ ID NO:35, 36 and 37 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:35, 36 and / or 37 are substituted with another amino acid), respectively (row 6); and FR1 , FR2, FR3 and / or FR4 having an amino acid sequence of SEQ ID NO:142, 143, 144 and 145 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:142, 143, 144 and / or 145 are substituted with another amino acid), respectively (row 6).

[0404] By way of further illustration, in some embodiments, the antigen-binding molecule comprises:

[0405] (i) a TCRa variable domain comprising: CDR1a, CDR2a and / or CDR3a having an amino acid sequence of SEQ ID NO:8, 9 and 10 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:8, 9 and / or 10 are substituted with another amino acid), respectively (row 2); and FR1 , FR2, FR3 and / or FR4 having an amino acid sequence of SEQ ID NO:106, 107, 108 and 109 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NQ:106, 107, 108 and / or 109 are substituted with another amino acid), respectively (row 2); in combination with,

[0406] (ii) a TCRp variable domain comprising: CDR1 p, CDR2p and / or CDR3p having an amino acid sequence of SEQ ID NO:11 , 12 and 13 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:11 , 12 and / or 13 are substituted with another amino acid), respectively (row 2); and FR1 , FR2, FR3 and / or FR4 having an amino acid sequence of SEQ ID NO:110, 111 , 112 and 113 (or a variant thereof in which 1 or 2 or 3 amino acids in SEQ ID NO:110, 111 , 112 and / or 113 are substituted with another amino acid), respectively (row 2). In some embodiments, the antigen-binding molecule comprises: (i) a TCRa variable domain having an amino acid sequence having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:60, 52, 50, 54, 56, 58, 62 or 64; in combination with (ii) a TCRp variable domain having an amino acid sequence having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:61 , 53, 51 , 55, 57, 59, 63 or 65.

[0407] In some embodiments, the antigen-binding molecule comprises: (i) a TCRa variable domain having an amino acid sequence having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the TCRa variable domain amino acid sequence as indicated in Column A of row 1 , 2, 3, 4, 5, 6, 7 or 8 of Table C; in combination with (ii) a TCRp variable domain having an amino acid sequence having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the TCRp variable domain amino acid sequence as indicated in row 1 , 2, 3, 4, 5, 6, 7 or 8 of Column B of Table C; wherein the TCRa variable domain amino acid sequence from Column A and the TCRp variable domain amino acid sequence from Column B are selected from the same row.

[0408] By way of illustration, in some embodiments, the antigen-binding molecule comprises: (i) a TCRa variable domain having an amino acid sequence having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NQ:60 (row 6); in combination with (ii) a TCRp variable domain having an amino acid sequence having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:61 (row 6).

[0409] By way of further illustration, in some embodiments, the antigen-binding molecule comprises: (i) a TCRa variable domain having an amino acid sequence having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:52 (row 2); in combination with (ii) a TCRp variable domain having an amino acid sequence having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:53 (row 2).

[0410] In any of the embodiments disclosed herein, an antigen-binding molecule / TCR or fragment thereof comprising an amino acid sequence described by reference to a given SEQ ID NO or variant thereof in which 1 or 2 or 3 amino acids are substituted with another amino acid, may instead comprise an amino acid sequence having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of the given SEQ ID NO.

[0411] In some embodiments described herein, one or more amino acids of an amino acid sequence referred to herein (e.g., an amino acid sequence of an antigen-binding molecule, e.g., an amino acid sequence of a CDR or variable domain) are substituted with another amino acid. A substitution comprises substitution of an amino acid residue with a non-identical ‘replacement’ amino acid residue. A replacement amino acid residue of a substitution according to the present disclosure may be a naturally-occurring amino acid residue ( / .e., encoded by the genetic code) which is non-identical to the amino acid residue at the relevant position of the equivalent, unsubstituted amino acid sequence, selected from: alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine (Cys), glutamine (Gin), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (lie): leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), and valine (Vai). In some embodiments, a replacement amino acid may be a non-naturally occurring amino acid residue - i.e., an amino acid residue other than those recited in the preceding sentence. Examples of non-naturally occurring amino acid residues include norleucine, ornithine, norvaline, homoserine, aib, and other amino acid residue analogues such as those described in Ellman, etal., Meth. Enzym. 202 (1991) 301-336.

[0412] In some embodiments, a substitution may be biochemically conservative. In some embodiments, where an amino acid to be substituted is provided in one of rows 1 to 5 of the table below, the replacement amino acid of the substitution is another, non-identical amino acid provided in the same row:

[0413] By way of illustration, in some embodiments wherein substitution is of a Met residue, the replacement amino acid may be selected from Ala, Vai, Leu, lie, Trp, Tyr, Phe and Norleucine.

[0414] In some embodiments, a replacement amino acid in a substitution may have the same side chain polarity as the amino acid residue it replaces. In some embodiments, a replacement amino acid in a substitution may have the same side chain charge (at pH 7.4) as the amino acid residue it replaces:

[0415] That is, in some embodiments, a nonpolar amino acid is substituted with another, non-identical nonpolar amino acid. In some embodiments, a polar amino acid is substituted with another, non-identical polar amino acid. In some embodiments, an acidic polar amino acid is substituted with another, non-identical acidic polar amino acid. In some embodiments, a basic polar amino acid is substituted with another, non- identical basic polar amino acid. In some embodiments, a neutral amino acid is substituted with another, non-identical neutral amino acid. In some embodiments, a positive amino acid is substituted with another, non-identical positive amino acid. In some embodiments, a negative amino acid is substituted with another, non-identical negative amino acid.

[0416] In some embodiments, substitution(s) may be functionally conservative. That is, in some embodiments, the substitution may not affect (or may not substantially affect) one or more functional properties (e.g., target binding) of the antigen-binding molecule comprising the substitution as compared to the equivalent unsubstituted molecule.

[0417] The TCRs according to the present disclosure may be synthetic / artificial TCRs comprising TCRa and TCRp human variable regions and mouse constant regions, for example to improve the expression of the TCR, and in order to use the mouse constant region for the tracking of transfected human T cells with an anti-mouse antibody.

[0418] Provided are human / mouse hybrid TCRs comprising mouse TCRa and TCRp constant region amino acid sequences of SEQ ID NO:167 and SEQ ID NO:168, respectively. Provided herein are also TCRs, e.g., synthetic or artificial TCRs, comprising TCRa and TCRp human variable regions and human constant regions. In some embodiments the human TCRa constant region is shown in SEQ ID NO:162. In some embodiments the human TCRp constant region is shown in SEQ ID NO:163 or 164.

[0419] In some embodiments, the TCR is a hybrid TCR construct. In some embodiments, the TCR comprises a human TCRa variable domain amino acid sequence and a mouse TCRa constant region. In some embodiments, the TCR comprises a human TCRp variable domain amino acid sequence and a mouse TCRp constant region. In some embodiments, the TCR comprises: (i) a human TCRa variable domain amino acid sequence and a mouse TCRa constant region; in combination with (ii) a human TCRp variable domain amino acid sequence and a mouse TCRp constant region.

[0420] In some embodiments, the TCRa constant region comprises an amino acid having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:167. In some embodiments, the TCRp constant region comprises an amino acid having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:168. It will be appreciated that such TCR constructs may further comprise any of the TCR variable domains described herein, e.g., human variable domains.

[0421] In some embodiments, the TCR is a fully human TCR construct. In some embodiments, the TCR comprises a human TCRa variable domain amino acid sequence and a human TCRa constant region. In some embodiments, the TCR comprises a human TCRp variable domain amino acid sequence and a human TCRp constant region. In some embodiments, the TCR comprises: (i) a human TCRa variable domain amino acid sequence and a human TCRa constant region; in combination with (ii) a human TCRp variable domain amino acid sequence and a human TCRp constant region.

[0422] In some embodiments, the TCRa constant region comprises an amino acid having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:162 or 165. In some embodiments, the TCRp constant region comprises an amino acid having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:163, 164, or 166. It will be appreciated that such TCR constructs may further comprise any of the TCR variable domains described herein, e.g., human variable domains.

[0423] In some embodiments, the TCRa constant region comprises an amino acid having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:162. In some embodiments, the TCRp constant region comprises an amino acid having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:163 or 164. It will be appreciated that such TCR constructs may further comprise any of the TCR variable domains described herein, e.g., human variable domains.

[0424] In some embodiments, any TCRa polypeptide / domain / chain and / or TCRp polypeptide / domain / chain described herein may comprise modifications to promote stable expression and / or reduce mismatching with endogenous TCR chains.

[0425] In some embodiments, any TCRa chain described herein may comprise a constant region comprising a cysteine residue at the amino acid position corresponding to position 48 of SEQ ID NO:162. In some embodiments, any TCRp chain described herein may comprise a constant region comprising a cysteine residue at the amino acid position corresponding to position 57 of SEQ ID NO:163 or 164. In some embodiments, any TCRa chain described herein may comprise a TCRa constant region comprising a cysteine residue at the amino acid position corresponding to position 48 of SEQ ID NO:162 and a TCRp constant region comprising a cysteine residue at the amino acid position corresponding to position 57 of SEQ ID NO:163 or 164.

[0426] In some embodiments, any TCRa chain described herein may comprise a constant region comprising a mutated version of the human TCRa constant region as in SEQ ID NO:165. In some embodiments, any TCRp chain described herein may comprise a constant region comprising a mutated version of the human TCRp constant region as in SEQ ID NO:166. In some embodiments, any TCR described herein may comprise a human TCR constant region comprising a mutated version of the human TCRa constant region as in SEQ ID NO:165 and a mutated version of human TCRp constant region as in SEQ ID NO:166. The mutation comprises the introduction of a Cys in both the alpha and beta chains of the TCR to create a stabilizing disulfide bridge between the two chains. TCR chains were modified by mutagenesis of residue 48 in the Ca region from Thr to Cys and residue 57 of the Cp region from Ser to Cys. The method has been described previously in Kuball et al, Blood. 2007 Mar 15; 109(6): 2331-2338., which is hereby incorporated by reference in its entirety. The mutation promotes stable expression and pairing of the transduced TCR in human T cells in which the endogenous TCR is not knocked out.

[0427] In some embodiments, an antigen-binding molecule according to the present disclosure, e.g., TCR or fragment thereof, comprises a TCRa chain comprising a TCRa constant region having at least 80%, 85%, 90%, or 95% sequence identity to an amino acid sequence selected from: SEQ ID NO:165, 162 and 167; and a TCRp chain comprising a TCRp constant region having at least 80%, 85%, 90%, or 95% sequence identity to an amino acid sequence selected from: SEQ ID NO:166, 163, 164 and 168.

[0428] In some embodiments, an antigen-binding molecule according to the present disclosure, e.g., TCR or fragment thereof, comprises a TCRa chain comprising a TCRa constant region having at least 80%, 85%, 90%, or 95% sequence identity to SEQ ID NO:165, and a TCRp chain comprising a TCRp constant region having at least 80%, 85%, 90%, or 95% sequence identity to SEQ ID NO:166.

[0429] In some embodiments, an antigen-binding molecule according to the present disclosure, e.g., TCR or fragment thereof, comprises a TCRa chain comprising a TCRa constant region having at least 80%, 85%, 90%, or 95% sequence identity to SEQ ID NO:162, and a TCRp chain comprising a TCRp constant region having at least 80%, 85%, 90%, or 95% sequence identity to SEQ ID NO:163 or 164. In some embodiments, any antigen-binding molecule, e.g., TCR or fragment thereof, described herein may be capable of recognising and / or binding to a peptide or antigen derived from WT-1 protein, i.e., a WT-1 -derived peptide or antigen, such as when presented by a major histocompatibility complex (MHC) molecule. In some embodiments, WT-1 comprises or consists of the amino acid sequence of SEQ ID NO:169, 184, 185, 186, 187, 188, 189 or 190. In some embodiments, WT-1 comprises or consists of the amino acid sequence of SEQ ID NO:169, 184, 185, 186, 188, or 189.

[0430] An ‘antigen’ refers to any molecule, e.g., a peptide, that provokes an immune response or is capable of being bound by a TCR. An ‘epitope,’ as used herein, refers to a portion of a polypeptide that provokes an immune response or is capable of being bound by a TCR. The immune response may involve either antibody production, or the activation of specific immunologically competent cells, or both. A person of skill in the art would readily understand that any macromolecule, including virtually all proteins or peptides, can serve as an antigen. An antigen and / or epitope can be endogenously expressed, i.e., expressed by genomic DNA, or can be recombinantly expressed. An antigen and / or epitope can be of exogenous origin. If of polypeptide origin, an antigen and / or epitope can possess modifications to the amino acids comprising the antigen and / or epitope {e.g., phosphorylation, glycosylation, cysteinylation, deamidation, and / or other post-translational modifications to the amino acids within the antigen and / or epitope). An antigen and / or epitope can be specific to a certain tissue or cell, such as a cancer cell, or it can be broadly expressed. In addition, fragments of larger molecules can act as antigens. In some embodiments herein, antigens are tumor antigens. An epitope can be present in a longer polypeptide {e.g., in a protein), or an epitope can be present as a fragment of a longer polypeptide. In some embodiments, an epitope is complexed with a major histocompatibility complex (MHC; also referred to herein as a HLA molecule, e.g., a HLA class I or class II molecule).

[0431] TCRs bind, via their CDR loops, to peptides presented by molecules of the major histocompatibility complex (MHC). This TCR-MHC interaction is crucially important in cell mediated immunity, with the specificity in the cellular immune response being attributable to MHC polymorphism, an extensive TCR repertoire, and a variable peptide cargo. The conventional T-cell response is mediated by TCR recognition of short peptide fragments bound to MHC class I or MHC class II molecules. Generally, MHC- I present peptides derived from endogenous proteins that are recognized by cytotoxic T-cells, whereas MHC-II present exogenously-derived peptides to T helper cells (see e.g., Bhati et al., Protein Science, 23:260-272 (2014), which is hereby incorporated by reference in its entirety).

[0432] The term ‘HLA,’ as used herein, refers to the human leukocyte antigen. In humans, MHCs are encoded by the human leukocyte antigen (HLA) locus on chromosome 6. There are three major HLA gene loci (HLA-A, HLA-B and HLA-C) and three minor loci (HLA-E, HLA-F and HLA-G). This locus is highly polymorphic, spans over 5 mega bases and covers over 200 genes. Individual subjects normally express 6 different classical MHC-I and 6 MHC-II molecules that can differ from each other by a single amino acid, or by more than 30 amino acids. These polymorphisms mostly affect the MHC binding cleft, and thus dictate the diversity of peptides presented by each MHC molecule (Bhati et al., Protein Science, 23:260- 272 (2014)). The frequency distribution of most common HLA alleles is publicly available (http: / / www.allelefrequencies.net / top10dist.asp). MHC proteins are expressed on the surface of cells and are involved in activation of the immune response. HLA class I genes encode MHC class I molecules, which are expressed on the surface of cells in complex with peptide fragments (antigens) of self or non-self proteins. T cells expressing TCR and CD3 recognize the antigemMHC class I complex and initiate an immune response to target and destroy antigen presenting cells displaying non-self proteins.

[0433] As used herein, an ‘HLA class I molecule’ or ‘MHC class I molecule’ refers to a protein product of a wildtype or variant HLA class I gene encoding an MHC class I molecule. Accordingly, ‘HLA class I molecule’ and ‘MHC class I molecule’ are used interchangeably herein.

[0434] MHC class I a-chains are polymorphic, and different a-chains are capable of binding and presenting different peptides. Genes encoding MHC class I a polypeptides are highly variable, with the result that cells from different subjects often express different MHC class I molecules.

[0435] The MHC Class I molecule comprises two protein chains: the alpha chain (a-chain) and the p2- microglobulin (p 2m) chain. Human p 2m is encoded by the B2M gene. The amino acid sequence of p 2m is set forth in SEQ ID NO:183. The alpha chain of the MHC Class I molecule is encoded by the HLA gene complex. The HLA complex is located within the 6p21 .3 region on the short arm of human chromosome 6 and contains more than 220 genes of diverse function. The HLA genes are highly variant, with over 36,000 HLA alleles and related alleles, including over 25,000 HLA Class I alleles, known in the art, encoding thousands of HLA proteins, including over 14,000 HLA Class I proteins (see, e.g., hla.alleles.org). There are at least three genes in the HLA complex that encode an MHC Class I alpha chain protein: HLA-A, HLA-B, and HLA-C. In addition, HLA-E, HLA-F, and HLA-G encode proteins that associate with the p 2m chain to form the MHC Class I molecule.

[0436] The term ‘antigen-presenting cell’, as used herein, designates cells having the capability to present processed antigenic moiety fragments via MHC class I or MHC class II molecules. Most cell types including cancer cells can express MHC class I molecules and present fragments via MHC class I molecules, while MHC class II molecules are expressed on professional antigen presenting cells. Professional antigen-presenting cells may be a B-cell, a monocyte, or a dendritic cell. The antigen presenting cells may be synthetic, or be isolated from peripheral blood mononuclear cells (PBMCs). Artificial APCs are a type of cell line that expresses a HLA molecule of interest for testing of TCR binding. The HLA protein can be endogenously expressed, or the artificial APCs can be engineered to express the HLA molecule of interest. Artificial APCs expressing the HLA allele of interest can be loaded with peptides such that the binding of a TCR to a peptide:HLA class I complex can be tested.

[0437] ‘Antigen-derived’, for example ‘WT-1 -derived’, refers to an immunogenic peptide / epitope being a portion of the antigen / polypeptide (e.g., WT-1) from which it has been processed. For example, an antigen is processed in the cell by the proteasome or immunoproteasome and the resulting antigen-derived peptides are presented on the MHC class I or MHC class II complex.

[0438] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, recognizes and / or binds to an antigen-derived peptide (e.g., a WT-1 -derived peptide).

[0439] In some embodiments, the WT-1-derived peptide is SEQ ID NO:1 , 179, 180, 181 , or 182. In some embodiments, the WT-1-derived peptide is SEQ ID NO:1. This peptide is located at positions 37 to 45 of SEQ ID NO:31 and may also be described as WT-I 37-45.

[0440] In some embodiments, the WT-1-derived peptide is SEQ ID NO:179, 180, 181 or 182.

[0441] In some embodiments, the WT-1-derived peptide is SEQ ID NO:179, wherein Xi is A or any amino acid other than A, X2 is G or any amino acid other than G, and / or X3 is A or any amino acid other than A.

[0442] In some embodiments, the WT-1-derived peptide is SEQ ID NQ:180, wherein Xi is A or any amino acid other than A, X2 is P or any amino acid other than P, X3 is G or any amino acid other than G, and / or X4 is A or any amino acid other than A.

[0443] In some embodiments, the WT-1-derived peptide is SEQ ID NO:181 , wherein Xi is L or any amino acid other than L, X2 is A or any amino acid other than A, X3 is P or any amino acid other than P, X4 is G or any amino acid other than G, and / or X5 is A or any amino acid other than A.

[0444] In some embodiments, the WT-1-derived peptide is SEQ ID NO:182, wherein Xi is A or any amino acid other than A, X2 is P or any amino acid other than P, and / or X3 is A or any amino acid other than A.

[0445] For conciseness, hereinbelow a peptide that is presented by an MHC class I molecule comprising an MHC class I a chain polypeptide encoded by a given HLA allele or a HLA allele within a given genus of HLA alleles may be referred to simply as being presented ‘through’ or ‘on’ the relevant allele. For example, a TCR that binds to a WT-1 -derived antigenic peptide presented by an MHC class I molecule comprising an MHC class I a chain polypeptide encoded by a HLA-A*02 allele may be described as a TCR that binds to a WT-1 -derived antigenic peptide presented through / on a HLA-A*02 allele.

[0446] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a WT-1 peptide, e.g., when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide encoded by HLA-A*02 (also known as HLA-A2, HLA-A02, and HLA-A*2). That is, when presented on HLA-A*02.

[0447] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a WT-1 peptide, e.g., when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide encoded by a HLA-A*02:01 allele. That is, when presented on HLA-A*02:01 .

[0448] In some embodiments, the antigen-binding molecule is capable of recognizing / binding, e.g., specifically binds, to a WT-1 -derived antigenic peptide presented through another HLA-A*02 allele, including but not restricted to: 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*02:11 , HLA-A*02:12, HLA-A*02:19, HLA-A*02:24, HLA-A*02:264, or HLA-A*02:52.

[0449] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a WT-1 peptide, e.g., when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide encoded by a HLA-A*02:07 allele. That is, when presented on HLA-A*02:07. In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a WT-1 peptide, e.g., when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide encoded by a HLA-A*02:02 allele. That is, when presented on HLA-A*02:02.

[0450] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to SEQ ID NO:1 , 179, 180, 181 or 182, e.g., when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide encoded by HLA-A*02 (also known as HLA-A2, HLA-A02, and HLA-A*2). That is, when presented on HLA-A*02.

[0451] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to SEQ ID NO: 1 , 179, 180, 181 or 182, e.g., when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide encoded by a HLA-A*02:01 allele, a HLA-A*02:07 allele, and / or a HLA-A*02:02 allele. That is, when presented on HLA-A*02:01 .

[0452] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to SEQ ID NO: 1 , 179, 180, 181 or 182, e.g., when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide encoded by a HLA-A*02:01 allele. That is, when presented on HLA-A*02:01.

[0453] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to SEQ ID NO: 1 , 179, 180, 181 or 182, e.g., when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide encoded by a HLA-A*02:07 allele. That is, when presented on HLA-A*02:07.

[0454] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to SEQ ID NO: 1 , 179, 180, 181 or 182, e.g., when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide encoded by a HLA-A*02:02 allele. That is, when presented on HLA-A*02:02.

[0455] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a VLDFAPPGA (SEQ ID NO:1)-HLA-A*02 complex. In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02:01 complex, a VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02:07 complex and / or a VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02:02 complex. In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02:01 complex. In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02:07 complex. In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02:02 complex.

[0456] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a VLDFXIPPX2X3 (SEQ ID NQ:179)-HLA-A*02 complex. In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a VLDFX1PPX2X3 (SEQ ID N0:179)-HLA-A*02:01 complex. In some embodiments, the antigen-binding molecule capable of recognizing / binding to a VLDFX1PPX2X3 (SEQ ID NO:179)-HLA-A*02 complex or a VLDFX1PPX2X3 (SEQ ID NQ:179)-HLA-A*02:01 complex is TCR_A0427.

[0457] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a VLDFX1X2PX3X4 (SEQ ID NQ:180)-HLA-A*02 complex. In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a VLDFX1X2PX3X4 (SEQ ID NQ:180)-HLA-A*02:01 complex. In some embodiments, the antigen-binding molecule capable of recognizing / binding to a VLDFX1X2PX3X4 (SEQ ID NQ:180)-HLA-A*02 complex or a VLDFX1X2PX3X4 (SEQ ID NQ:180)-HLA-A*02:01 complex is TCR_A0429.

[0458] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a VX1DFX2X3PX4X5 (SEQ ID NQ:181)-HLA-A*02 complex. In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a VXIDFX2X3PX4X5 (SEQ ID NQ:181)-HLA-A*02:01 complex. In some embodiments, the antigen-binding molecule capable of recognizing / binding to a VXI DFX2X3PX4X5 (SEQ ID NQ:181)-HLA-A*02 complex or a VX1DFX2X3PX4X5 (SEQ ID NQ:181)-HLA-A*02:01 complex is TCR_A0431.

[0459] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a VLDFX1PX2GX3 (SEQ ID NQ:182)-HLA-A*02 complex. In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a VLDFX1PX2GX3 (SEQ ID NQ:182)-HLA-A*02:01 complex. In some embodiments, the antigen-binding molecule capable of recognizing / binding to a VLDFXI PX2GX3 (SEQ ID NQ:182)-HLA-A*02 complex or a VLDFX1PX2GX3 (SEQ ID NQ:182)-HLA-A*02:01 complex is TCR_A0432.

[0460] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a peptide :MHC complex comprising an MHC class I a chain polypeptide encoded by a HLA-A*02 allele (e.g., HLA-A*02:01 , HLA-A*02:07, HLA-A*02:02), and a peptide comprising or consisting of SEQ ID NO:1 , 179, 180, 181 or 182.

[0461] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a peptide :MHC complex comprising an MHC class I a chain polypeptide encoded by a HLA-A*02 allele (e.g., HLA-A*02:01 , HLA-A*02:07, HLA-A*02:02), and a peptide comprising or consisting of VLDFAPPGA (SEQ ID NO:1).

[0462] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a peptide:MHC complex comprising an MHC class I a chain polypeptide encoded by a HLA-A*02 allele (e.g., HLA-A*02:01), and a peptide comprising or consisting of VLDFX1PPX2X3 (SEQ ID NO:179). In some embodiments, the peptide comprises or consists of SEQ ID NO:179, wherein Xi is A or any amino acid other than A, X2 is G or any amino acid other than G, and / or X3 is A or any amino acid other than A.

[0463] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a peptide:MHC complex comprising an MHC class I a chain polypeptide encoded by a HLA-A*02 allele {e.g., HLA-A*02:01), and a peptide comprising or consisting of VLDFX1X2PX3X4 (SEQ ID NO:180). In some embodiments, the peptide comprises or consists of SEQ ID NO:180, wherein Xi is A or any amino acid other than A, X2 is P or any amino acid other than P, X3 is G or any amino acid other than G, and / or X4 is A or any amino acid other than A.

[0464] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a peptide :MHC complex comprising an MHC class I a chain polypeptide encoded by a HLA-A*02 allele {e.g., HLA-A*02:01), and a peptide comprising or consisting of VX1DFX2X3PX4X5 (SEQ ID NO:181). In some embodiments, the peptide comprises or consists of SEQ ID NO:181 , wherein Xi is L or any amino acid other than L, X2 is A or any amino acid other than A, X3 is P or any amino acid other than P, X4 is G or any amino acid other than G, and / or X5 is A or any amino acid other than A.

[0465] In some embodiments, an antigen-binding molecule as described herein, e.g., a TCR or fragment thereof, is capable of recognizing / binding to a peptide:MHC complex comprising an MHC class I a chain polypeptide encoded by a HLA-A*02 allele {e.g., HLA-A*02:01), and a peptide comprising or consisting of VLDFX1PX2GX3 (SEQ ID NO:182). In some embodiments, the peptide comprises or consists of SEQ ID NO:182, wherein Xi is A or any amino acid other than A, X2 is P or any amino acid other than P, and / or X3 is A or any amino acid other than A.

[0466] WT-1 peptide VLDFAPPGA (SEQ ID NO:1) has been reported as an immunogenic epitope presented on HLA-A*02:01 and TCRs against this peptide have been discovered previously, see for example US11597755B2, which is hereby incorporated by reference in its entirety.

[0467] In some embodiments, an antigen-binding molecule, e.g., a TCR, described herein comprises a TCRa variable domain having less than 95% amino acid sequence identity, e.g., one of <94%, <93%, <92%, <91 %, <90%, <85%, <80%, <75%, <70%, <65%, <50%, <55%, <40%, <35%, <30% or <25% amino acid sequence identity, with a TCRa variable domain amino acid sequence disclosed in US11597755B2 {e.g., a TCRa variable domain sequence having the amino acid sequence of SEQ ID NO:4, 37, 93, 267, 104, 109, 114, 278 or 215 from US11597755B2). In some embodiments, an antigen-binding molecule, e.g., a TCR, described herein does not comprise a TCRa variable domain having at least at least 25% amino acid sequence identity, e.g., one of >30%, >35%, >40%, >45%, >50%, >55%, >60%, >65%, >70%, >75%, >80%, >85%, >90%, >91 %, >92%, >93%, >94% or 95% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:4, 37, 93, 267, 104, 109, 114, 278 or 215 from US11597755B2.

[0468] In some embodiments, an antigen-binding molecule, e.g., a TCR, described herein comprises a TCRp variable domain having less than 95% amino acid sequence identity, e.g., one of <94%, <93%, <92%, <91 %, <90%, <85%, <80%, <75%, <70%, <65%, <50%, <55%, <40%, <35%, <30% or <25% amino acid sequence identity, with a TCRa variable domain amino acid sequence disclosed in US11597755B2 {e.g., a TCRp variable domain sequence having the amino acid sequence of SEQ ID NO:9, 42, 98, 272, 164, 170, 283, 289 or 220 from US11597755B2). In some embodiments, an antigen-binding molecule, e.g., a TCR, described herein does not comprise a TCRp variable domain having at least at least 25% amino acid sequence identity, e.g., one of >30%, >35%, >40%, >45%, >50%, >55%, >60%, >65%, >70%, >75%, >80%, >85%, >90%, >91%, >92%, >93%, >94% or 95% amino acid sequence identity, with the amino acid sequence of SEQ ID NO: 9, 42, 98, 272, 164, 170, 283, 289 or 220 from US11597755B2.

[0469] The TCRs / antigen-binding molecules described herein may be comprised within multispecific molecules. That is, the present invention provides a multispecific antigen-binding molecule comprising an antigenbinding molecule e.g., a TCR or an antigen-binding fragment thereof described herein. A multispecific antigen-binding molecule may be e.g., bispecific, trispecific, etc. Also provided is the use of a TCR / antigen-binding molecule described herein as part of a fusion construct, wherein said fusion construct comprises a TCR / antigen-binding molecule described herein and an antigen-binding molecule that is capable of binding to a molecule expressed {e.g., specifically expressed) by immune cells, e.g., T cells, including but not limited to CD3.

[0470] In some embodiments, a multispecific antigen-binding molecule according to the present disclosure comprises an antigen-binding molecule e.g., a TCR or an antigen-binding fragment thereof described herein and at least one further antigen-binding molecule that is capable of binding to a molecule expressed by an immune cell. The two or more antigen-binding molecules may be expressed / presented as a fusion construct or fusion protein.

[0471] In some embodiments, a multispecific antigen-binding molecule according to the present disclosure comprises an antigen-binding molecule e.g., a TCR or an antigen-binding fragment thereof described herein and at least one further antigen-binding molecule that is capable of binding to a molecule expressed by a T cell. The molecule expressed by an immune cell / a T cell may be CD3. The molecule expressed by an immune cell / a T cell may be CD4 or CD8. That is, the TCRs / antigen-binding molecules described herein may be used in bispecific T cell engagers (BiTEs) or T cell Engaging Receptors (TCER®). Such multispecific molecules can target immune cells, e.g., via a CD3-binding arm, to diseased cells that are recognised via the TCR moiety.

[0472] In some embodiments, the TCR / antigen-binding molecule described herein and / or the at least one further antigen-binding molecule that is capable of binding to a molecule expressed by an immune cell are single-chain molecules, e.g., in scFv form. The at least two antigen-binding molecules may be connected via a linker.

[0473] Functional properties of the TCRs

[0474] The antigen-binding molecules / TCRs / TCR chains / TCR fragments described herein may be characterised by reference to certain functional properties. In some embodiments, an article described herein may possess one or more of the following properties: recognizes and / or binds {e.g., specifically binds) to WT-1 ; recognizes and / or binds {e.g., specifically binds) to a WT-1 -derived peptide {e.g., an antigenic peptide); recognizes and / or binds {e.g., specifically binds) to VLDFAPPGA (SEQ ID NO:1); recognizes and / or binds {e.g., specifically binds) to VLDFX1PPX2X3 (SEQ ID NO:179); recognizes and / or binds (e.g., specifically binds) to VLDFX1X2PX3X4 (SEQ ID NO:180); recognizes and / or binds (e.g., specifically binds) to VX1DFX2X3PX4X5 (SEQ ID NO:181); recognizes and / or binds e.g., specifically binds) to VLDFX1PX2GX3 (SEQ ID NO:182); recognizes and / or binds (e.g., specifically binds) to a WT-1 -derived peptide (e.g., an antigenic peptide) when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide, e.g., encoded by HLA-A*02, e.g., encoded by HLA-A*02:01 ; recognizes and / or binds (e.g., specifically binds) to VLDFAPPGA (SEQ ID NO:1) when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide, e.g., encoded by HLA-A*02, e.g., encoded by HLA-A*02:01 ; recognizes and / or binds (e.g., specifically binds) to VLDFX1PPX2X3 (SEQ ID NO:179) when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide, e.g., encoded by HLA-A*02, e.g., encoded by HLA-A*02:01 ; recognizes and / or binds (e.g., specifically binds) to VLDFX1X2PX3X4 (SEQ ID NQ:180) when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide, e.g., encoded by HLA-A*02, e.g., encoded by HLA-A*02:01 ; recognizes and / or binds (e.g., specifically binds) to VX1DFX2X3PX4X5 (SEQ ID NO:181) when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide, e.g., encoded by HLA-A*02, e.g., encoded by HLA-A*02:01 ; recognizes and / or binds (e.g., specifically binds) to VLDFX1PX2GX3 (SEQ ID NO:182) when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide, e.g., encoded by HLA-A*02, e.g., encoded by HLA-A*02:01 ; recognizes and / or binds (e.g., specifically binds) to a cell expressing WT-1 ; recognizes and / or binds (e.g., specifically binds) to a cell expressing a WT-1 -derived peptide (e.g., an antigenic peptide); recognizes and / or binds (e.g., specifically binds) to a cell presenting VLDFAPPGA (SEQ ID NO:1), e.g., when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide, e.g., encoded by HLA-A*02, e.g., encoded by HLA-A*02:01 ; recognizes and / or binds (e.g., specifically binds) to a cell presenting VLDFX1PPX2X3 (SEQ ID NO:179), e.g., when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide, e.g., encoded by HLA-A*02, e.g., encoded by HLA-A*02:01 ; recognizes and / or binds (e.g., specifically binds) to a cell presenting VLDFX1X2PX3X4 (SEQ ID NQ:180), e.g., when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide, e.g., encoded by HLA-A*02, e.g., encoded by HLA-A*02:01 ; recognizes and / or binds (e.g., specifically binds) to a cell presenting VX1DFX2X3PX4X5 (SEQ ID NO:181), e.g., when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide, e.g., encoded by HLA-A*02, e.g., encoded by HLA-A*02:01 ; recognizes and / or binds (e.g., specifically binds) to a cell presenting VLDFX1PX2GX3 (SEQ ID NO:182), e.g., when presented by a major histocompatibility complex (MHC) molecule comprising an MHC class I a chain polypeptide, e.g., encoded by HLA-A*02, e.g., encoded by HLA-A*02:01 ; activates a host cell, e.g., T cell, in which the article is comprised / expressed; promotes cell killing activity of a cell in which the article is comprised / expressed; has an ‘anti-cancer’ effect when comprised / expressed in a host cell, e.g., T cell; has an ‘anti-tumor’ effect when comprised / expressed in a host cell, e.g., T cell; confers cytotoxic activity when comprised / expressed in a host cell, e.g., T cell; and / or promotes IFNy secretion from a host cell, e.g., T cell, in which it is comprised / expressed.

[0475] It will be appreciated that a given antigen-binding molecule may display more than one of the properties recited in the preceding paragraph. A given antigen-binding molecule may be evaluated for the properties recited in the preceding paragraph using suitable assays. For example, the assays may be e.g., in vitro assays, optionally cell-based assays or cell-free assays. In some embodiments, the assays may be e.g., in vivo assays, i.e., performed in non-human animals. In some embodiments, the assays may be e.g., ex vivo assays, i.e., performed using cells / tissue / an organ obtained from a subject.

[0476] Where assays are cell-based assays, they may comprise treating cells with a given host cell comprising / expressing the antigen-binding molecule / TCR in order to determine whether the host cells / antigen-binding molecule / TCR displays one or more of the recited properties. Assays may employ species labelled with detectable entities in order to facilitate their detection. Assays may comprise evaluating the recited properties following treatment of cells separately with a range of quantities / concentrations of a given host cell / antigen-binding molecule / TCR (e.g., a dilution series). It will be appreciated that the cells preferably express the target antigen for the antigen-binding molecule / TCR (i.e., a WT-1 -derived peptide, such as those described herein).

[0477] The ability of an antigen-binding molecule to bind specifically to a given molecule / complex can be determined by analysis according to methods known in the art, such as by ELISA, Surface Plasmon Resonance (SPR; see e.g., Hearty et al., Methods Mol Biol (2012) 907:411-442), Bio-Layer Interferometry (BLI; see e.g., Lad et al., (2015) J Biomol Screen 20(4): 498-507), flow cytometry, or by a radiolabelled antigen-binding assay (RIA) enzyme-linked immunosorbent assay. Through such analysis, binding to a given molecule can be measured and quantified. In some embodiments, the binding may be the response detected in a given assay.

[0478] The term ‘specifically binds,’ as used herein with respect to a T cell receptor, refers to a T cell receptor which recognizes a specific antigen complexed with an MHC molecule, but does not substantially recognize or bind other antigemMHC complexes in a sample.

[0479] Cell killing can be investigated, for example, using any of the methods reviewed in Zaritskaya et al., Expert Rev Vaccines (2011), 9(6):601-616, hereby incorporated by reference in its entirety. Examples of in vitro assays of cytotoxicity / cell killing assays include release assays such as the51Cr release assay, the lactate dehydrogenase (LDH) release assay, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) release assay, ATP release assay using Cell Titre Gio, and the calcein-acetoxymethyl (calcein-AM) release assay. These assays measure cell killing based on the detection of factors released from lysed cells.

[0480] An ‘anti-cancer effect’ as used herein, refers to a biological effect that can present as an increase in killing of cancer cells, a decrease in the number of cancer cells, a decrease in cancer cell proliferation, a decrease in the number / volume of metastases, an increase in overall or progression-free survival, an increase in life expectancy or amelioration of various physiological symptoms associated with the cancer. An anti-cancer effect can also refer to the prevention of the occurrence of a cancer, e.g., a vaccine. An ‘anti-tumor effect’ as used herein, refers to a biological effect that can present as an increase in killing of tumor cells, a decrease in tumor volume, a decrease in the number of tumor cells, a decrease in tumor cell proliferation, a decrease in the number of metastases, an increase in overall or progression-free survival, an increase in life expectancy, or amelioration of various physiological symptoms associated with the tumor. An anti-tumor effect can also refer to the prevention of the occurrence of a tumor, e.g., a vaccine.

[0481] An antigen-binding molecule of the present disclosure may be analysed for the properties described in the preceding paragraph in appropriate assays. Such assays include e.g., in vivo models.

[0482] In some embodiments, the functionality of TCRs can be assessed by quantification of cytokine secretion in the cell culture media.

[0483] It is well known that cytokines and their signaling pathways exert potent effects on T cell activation, differentiation, and function. Interferon gamma (IFNy) is crucial for Th1 differentiation and induction of IFNy release. In other subsets, IFNy inhibits the differentiation of Th2 and Th17 cells but has been shown to promote Regulatory T cells (Tregs) and antigen-specific memory T cell generation (Bishop et al. Front Immunol. 2021 Apr 13). IFNy is a key moderator of cell-mediated immunity with diverse, mainly pro- inflammatory actions on immunocytes and target tissue. Recent studies have shown it may enhance antitumor and antiviral effects of CD8 T cells. IFNy is released in large amounts by macrophages, activated CD8 T cells, natural killer T cells, and Th1 CD4 T cells (Bhat et al. Cell Death Dis. 2017 Jun

[0484] 1 ;8(6):e2836). IFNy secreted by T cells into the culture medium during the cytotoxicity assay can be quantified by ELISA methodology.

[0485] The functionality of TCRs of the present disclosure can be assessed using primary human T cells transduced with said TCRs.

[0486] Peripheral blood mononuclear cells (PBMCs) from healthy donors can be used as a source of primary T cells for the preparation of effector cells, e.g., as described herein. Transduction efficiency can be verified by flow cytometry, e.g., using an anti-mouse TCR antibody to verify that at least 50% of T cells express the transduced TCR.

[0487] The functionality of TCRs can be assessed by quantification of cytotoxicity using a flow cytometry assay, e.g., as described herein. For example, TCR-expressing T cells can be used as effector cells and can be mixed with cells expressing the target peptide (target cells).

[0488] The functionality of TCRs can also be assessed by quantification of cytotoxicity, e.g., using xCelligence impedance readout as described herein.

[0489] Nucleic acids and vectors

[0490] Any antigen-binding molecule, e.g., TCR, multispecific antigen-binding molecule or fragment thereof, according to the present disclosure may be encoded by any nucleotide sequence that encodes for the required amino acid sequence(s), taking into account codon degeneracy. The present disclosure provides nucleic acids, or a plurality of nucleic acids, encoding the TCRs, antigenbinding molecules, polypeptides and polypeptide complexes according to the present disclosure. In some embodiments, the nucleic acid(s) comprise or consist of DNA and / or RNA. In some embodiments, the nucleic acid is a polynucleotide, e.g., a polydeoxyribonucleotide or a polyribonucleotide.

[0491] An antigen-binding molecule or polypeptide according to the present disclosure may be produced within a cell by translation of RNA encoding the polypeptide(s). An antigen-binding molecule or polypeptide according to the present disclosure may be produced within a cell by transcription from nucleic acid encoding the polypeptide(s), and subsequent translation of the transcribed RNA.

[0492] In some embodiments, the nucleic acid(s) may be, or may be comprised / contained in, a vector, or a plurality of vectors.

[0493] Accordingly, the present disclosure also provides vectors, plasmids, and a plurality of vectors and / or plasmids containing nucleic acids, or a plurality of nucleic acids, according to the present disclosure. Nucleic acids, nucleotide sequences, plasmids and vectors according to the present disclosure may be provided in purified or isolated form, i.e., from other nucleic acids, plasmids, vectors or naturally-occurring biological material.

[0494] Provided herein is a nucleotide sequence encoding a TCRa variable domain according to the present disclosure. In some embodiments, the nucleotide sequence encoding a TCRa variable domain has at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:76, 92, 68, 84, 66, 82, 70, 86, 72, 88, 74, 90, 78, 94,

[0495] 80 or 96. In some embodiments, the nucleotide sequence encoding a TCRa variable domain has at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO: 76 or 92. In some embodiments, the nucleotide sequence encoding a TCRa variable domain has at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:68 or 84.

[0496] Provided herein is a nucleotide sequence encoding a TCRp variable domain according to the present disclosure. In some embodiments, the nucleotide sequence encoding a TCRp variable domain has at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:77, 93, 69, 85, 67, 83, 71 , 87, 73, 89, 75, 91 , 79, 95,

[0497] 81 or 97. In some embodiments, the nucleotide sequence encoding a TCRp variable domain has at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:77 or 93. In some embodiments, the nucleotide sequence encoding a TCRp variable domain has at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:69 or 85.

[0498] In some embodiments, the antigen-binding molecule, e.g., TCR, of the present disclosure comprises a polypeptide or polypeptides encoded by a nucleic acid / nucleotide sequence described herein.

[0499] In some embodiments, the antigen-binding molecule, e.g., TCR, of the present disclosure comprises a TCRa variable domain encoded by a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:76, 92, 68, 84, 66, 82, 70, 86, 72, 88, 74, 90, 78, 94, 80 or 96. In some embodiments, the antigen-binding molecule, e.g., TCR, of the present disclosure comprises a TCRa variable domain encoded by a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:76 or 92. In some embodiments, the antigen-binding molecule, e.g., TCR, of the present disclosure comprises a TCRa variable domain encoded by a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:68 or 84.

[0500] In some embodiments, the antigen-binding molecule, e.g., TCR, of the present disclosure comprises a TCRp variable domain encoded by a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:77, 93, 69, 85, 67, 83, 71 , 87, 73, 89, 75, 91 , 79, 95, 81 or 97. In some embodiments, the antigen-binding molecule, e.g., TCR, of the present disclosure comprises a TCRp variable domain encoded by a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:77 or 93. In some embodiments, the antigen-binding molecule, e.g., TCR, of the present disclosure comprises a TCRp variable domain encoded by a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:69 or 85.

[0501] In some embodiments, the antigen-binding molecule, e.g., TCR, of the present disclosure comprises: (i) a TCRa variable domain encoded by a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:76, 92, 68, 84, 66, 82, 70, 86, 72, 88, 74, 90, 78, 94, 80 or 96; in combination with (ii) a TCRp variable domain encoded by a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:77, 93, 69, 85, 67, 83, 71 , 87, 73, 89, 75, 91 , 79, 95, 81 or 97.

[0502] In some embodiments, the antigen-binding molecule, e.g., TCR, of the present disclosure comprises: (i) a TCRa variable domain encoded by a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the TCRa variable domain encoded by a nucleotide sequence as indicated in Column A of row 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15 or 16 of Table D; in combination with (ii) a TCRp variable domain encoded by a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the TCRp variable domain encoded by a nucleotide sequence as indicated in Column B of row 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15 or 16 of Table D; wherein the nucleotide sequence encoding the TCRa variable domain from Column A and the nucleotide sequence encoding the TCRp variable domain from Column B are selected from the same row.

[0503] By way of illustration, in some embodiments, the antigen-binding molecule, e.g., TCR, of the present disclosure comprises: (i) a TCRa variable domain encoded by a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:92 (row 12); and (ii) a TCRp variable domain encoded by a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:93 (row 12).

[0504] By way of further illustration, in some embodiments, the antigen-binding molecule, e.g., TCR, of the present disclosure comprises: (i) a TCRa variable domain encoded by a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO: 84 (row 4); and (ii) a TCRp variable domain encoded by a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:85 (row 4).

[0505] In some embodiments, the antigen-binding molecule, e.g., TCR, of the present disclosure comprises: (i) a TCRa variable domain encoded by a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of as indicated in Column A of row 1 , 3, 5, 7, 9, 1 1 , 13 or 15 of Table D; in combination with (ii) a TCRp variable domain encoded by a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the TCRp variable domain encoded by a nucleotide sequence as indicated in Column B of row 2, 4, 6, 8, 10, 12, 14 or 16 of Table D, respectively. In some embodiments, the antigen-binding molecule, e.g., TCR, of the present disclosure comprises: (i) a TCRa variable domain encoded by a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of as indicated in Column A of row 2, 4, 6, 8, 10, 12, 14 or 16 of Table D; in combination with (ii) a TCRp variable domain encoded by a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the TCRp variable domain encoded by a nucleotide sequence as indicated in Column B of row 1 , 3, 5, 7, 9, 11 , 13 or 15 of Table D, respectively.

[0506] Any of the nucleotide sequences described herein may be replaced by a codon degenerate nucleotide sequence thereof encoding the amino acid sequence encoded by the reference sequence.

[0507] As used herein, the term ‘nucleic acid’ refers to a polymer comprising multiple nucleotide monomers (e.g., ribonucleotide monomers or deoxyribonucleotide monomers). ‘Nucleic acid’ includes, for example, genomic DNA, cDNA, RNA, and DNA-RNA hybrid molecules. Nucleic acid molecules can be naturally occurring, recombinant, or synthetic. In addition, nucleic acid molecules can be single- stranded, doublestranded or triple- stranded. In some embodiments, nucleic acid molecules can be modified. In the case of a double-stranded polymer, ‘nucleic acid’ can refer to either or both strands of the molecule.

[0508] The term ‘nucleotide sequence,’ in reference to a nucleic acid, refers to a contiguous series of nucleotides that are joined by covalent linkages, such as phosphorus linkages (e.g., phosphodiester, alkyl and aryl- phosphonate, phosphorothioate, phosphotriester bonds), and / or non-phosphorus linkages (e.g., peptide and / or sulfamate bonds).

[0509] The terms ‘nucleotide’ and ‘nucleotide monomer’ refer to naturally occurring ribonucleotide or deoxyribonucleotide monomers, as well as non-naturally occurring derivatives and analogs thereof. Accordingly, nucleotides can include, for example, nucleotides comprising naturally occurring bases (e.g., adenosine, thymidine, guanosine, cytidine, uridine, inosine, deoxyadenosine, deoxythymidine, deoxyguanosine, or deoxycytidine) and nucleotides comprising modified bases known in the art.

[0510] Where a nucleotide sequence is disclosed herein, the reverse complement thereof is also expressly contemplated. Moreover, in each instance wherein a nucleotide sequence is disclosed herein, codon degenerate nucleotide sequences thereof encoding the same amino acid sequence are also expressly contemplated. A ‘codon degenerate nucleotide sequence’ of a reference nucleotide sequence refers to a nucleotide sequence having a non-identical nucleotide sequence to the nucleotide sequence of the reference nucleotide sequence, but encoding the same amino acid sequence as the amino acid sequence encoded by the reference nucleotide sequence, as a consequence of degeneracy of the genetic code.

[0511] ‘Encoding’ refers to the inherent property of specific sequences of nucleotides in a polynucleotide, such as a gene, a cDNA, or an mRNA, to serve as templates for synthesis of other polymers and macromolecules in biological processes having either a defined sequence of nucleotides (i.e., rRNA, tRNA and mRNA) or a defined sequence of amino acids and the biological properties resulting therefrom. Thus, a gene encodes a protein if transcription and translation of mRNA corresponding to that gene produces the protein in a cell or other biological system. Both the coding strand, the nucleotide sequence of which is identical to the mRNA sequence and is usually provided in sequence listings, and the noncoding strand, used as the template for transcription of a gene or cDNA, can be referred to as encoding the protein or other product of that gene or cDNA.

[0512] Unless otherwise specified, a ‘nucleotide sequence encoding an amino acid sequence’ includes all nucleotide sequences that are degenerate versions of each other and that encode the same amino acid sequence. The phrase nucleotide sequence that encodes a protein or an RNA may also include introns to the extent that the nucleotide sequence encoding the protein may in some version contain an intron(s).

[0513] In various embodiments, a nucleotide sequence according to the present disclosure encoding one or more antigen-binding molecules or polypeptides of the TCR is codon optimized, e.g., for expression in a chosen cell, such as a mammalian cell. The mammalian cell may be an immune cell, such as a T cell, e.g., a human T cell.

[0514] Codon optimization is a common method used to increase the expression of recombinant proteins, especially in the field of biotherapeutics. Its basis lies in the use of synonymous codon mutations in messenger RNA (mRNA) coding regions. Codon optimization is known to maximize protein expression by overcoming expression limitations associated with codon usage. This routine method has been reported to increase protein expression by up to >1 OOO-fold. This method is often applied in order to fine-tune the expression of one of two light chain genes of a bispecific antibody (Mauro, BioDrugs 32;69-81 (2018)).

[0515] Altering codon usage is possible since the 20 amino acids are encoded by 61 codons. Except for methionine and tryptophan, which are encoded by a single codon each, all other amino acids are specified by two to six redundant codons. Synonymous codon usage is not random, as it varies between different organisms, between different tissues of the same organism, and even between different parts of the same gene (Mauro, BioDrugs 32;69-81 (2018)).

[0516] Provided herein is a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:92, 84, 82, 86, 88, 90, 94 or 96. Said nucleotide sequences encode TCRa variable domains and are codon- optimised sequences derived from SEQ ID NO:76, 68, 66, 70, 72, 74, 78 or 80, respectively. That is, SEQ ID NO:76, 68, 66, 70, 72, 74, 78 and 80 are codon-optimised to produce SEQ ID NO: 92, 84, 82, 86, 88, 90, 94 or 96, respectively.

[0517] Provided herein is a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:92. Said nucleotide sequence encodes a TCRa variable domain and is a codon-optimised sequence derived from SEQ ID NO:76. That is, SEQ ID NO:76 is codon-optimised to produce SEQ ID NO:92.

[0518] Provided herein is a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:84. Said nucleotide sequence encodes a TCRa variable domain and is a codon-optimised sequence derived from SEQ ID NO:68. That is, SEQ ID NO:68 is codon-optimised to produce SEQ ID NO:84.

[0519] Provided herein is a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:93, 85, 83, 87, 89, 91 , 95 or 97. Said nucleotide sequences encode TCRp variable domains and are codon- optimised sequences derived from SEQ ID NO:77, 69, 67, 71 , 73, 89, 91 , 79 or 81 , respectively. That is, SEQ ID NO: 77, 69, 67, 71 , 73, 89, 91 , 79 and 81 are codon-optimised to produce SEQ ID NO: 93, 85, 83, 87, 89, 91 , 95 and 97, respectively.

[0520] Provided herein is a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:93. Said nucleotide sequence encodes a TCRp variable domain and is a codon-optimised sequence derived from SEQ ID NO:77. That is, SEQ ID NO:77 is codon-optimised to produce SEQ ID NO:93.

[0521] Provided herein is a nucleotide sequence having at least 70% nucleotide sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% nucleotide sequence identity, with the nucleotide sequence of SEQ ID NO:85. Said nucleotide sequence encodes a TCRp variable domain and is a codon-optimised sequence derived from SEQ ID NO:69. That is, SEQ ID NO:69 is codon-optimised to produce SEQ ID NO:85.

[0522] In some embodiments, a TCR according to the present disclosure may be expressed as a hybrid TCR construct. In some embodiments, a TCR according to the present disclosure may be expressed as a TCR construct comprising a human TCRa variable domain amino acid sequence and a mouse TCRa constant region. In some embodiments, a TCR according to the present disclosure may be expressed as a TCR construct comprising a human TCRp variable domain amino acid sequence and a mouse TCRp constant region. In some embodiments, a TCR according to the present disclosure may be expressed as a TCR construct comprising (i) a human TCRa variable domain amino acid sequence and a mouse TCRa constant region; in combination with (ii) a human TCRp variable domain amino acid sequence and a mouse TCRp constant region.

[0523] In some embodiments, a TCR according to the present disclosure may be expressed as a TCR construct wherein the TCRa constant region comprises an amino acid having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:167. In some embodiments, a TCR according to the present disclosure may be expressed as a TCR construct wherein the TCRp constant region comprises an amino acid having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:168. In some embodiments, a TCR according to the present disclosure may be expressed as a fully human TCR construct. In some embodiments, a TCR according to the present disclosure may be expressed as a TCR construct comprising a human TCRa variable domain amino acid sequence and a human TCRa constant region. In some embodiments, a TCR according to the present disclosure may be expressed as a TCR construct comprising a human TCRp variable domain amino acid sequence and a human TCRp constant region. In some embodiments, a TCR according to the present disclosure may be expressed as a TCR construct comprising (i) a human TCRa variable domain amino acid sequence and a human TCRa constant region; in combination with (ii) a human TCRp variable domain amino acid sequence and a human TCRp constant region.

[0524] In some embodiments, a TCR according to the present disclosure may be expressed as a TCR construct wherein the TCRa constant region comprises an amino acid having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:162 or 165. In some embodiments, a TCR according to the present disclosure may be expressed as a TCR construct wherein the TCRp constant region comprises an amino acid having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:163, 164, or 166.

[0525] In some embodiments, a TCR according to the present disclosure may be expressed as a TCR construct wherein the TCRa constant region comprises an amino acid having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:162. In some embodiments, a TCR according to the present disclosure may be expressed as a TCR construct wherein the TCRp constant region comprises an amino acid having at least 70% amino acid sequence identity, e.g., one of >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity, with the amino acid sequence of SEQ ID NO:163 or 164.

[0526] In some embodiments, a TCR according to the present disclosure may be expressed as a TCR construct wherein the TCRa polypeptide / domain / chain and / or TCRp polypeptide / domain / chain comprise a modification(s) capable of promoting stable expression of the TCR and / or reducing mismatching with endogenous TCR polypeptides / domains / chains.

[0527] In some embodiments, a TCR according to the present disclosure may be expressed as a TCR construct wherein the TCRa constant region comprises a cysteine residue at the amino acid position corresponding to position 48 of SEQ ID NO:162. In some embodiments, a TCR according to the present disclosure may be expressed as a TCR construct wherein the TCRp constant region comprises a cysteine residue at the amino acid position corresponding to position 57 of SEQ ID NO:163 or 164. In some embodiments, a TCR according to the present disclosure may be expressed as a TCR construct wherein the TCRa constant region comprises a cysteine residue at the amino acid position corresponding to position 48 of SEQ ID NO:162 and the TCRp constant region comprises a cysteine residue at the amino acid position corresponding to position 57 of SEQ ID NO:163 or 164. In some embodiments, a TCR according to the present disclosure may be expressed as a TCR construct wherein the TCRa constant region comprises an amino acid having the sequence of SEQ ID NO:165 and the TCRp constant region comprises an amino acid having the sequence of SEQ ID NO:166.

[0528] An antigen-binding molecule, TCR or polypeptide according to the present disclosure may be produced within a cell by translation of RNA encoding the relevant polypeptide(s). An antigen-binding molecule, TCR or polypeptide according to the present disclosure may be produced within a cell by transcription from nucleic acid(s) encoding the relevant polypeptide(s), and subsequent translation of the transcribed RNA. Constituent polypeptides of a TCR or antigen-binding molecule according to the present disclosure may be encoded by different nucleic acids or plurality of nucleic acids, or by different vectors or plurality of vectors.

[0529] In some embodiments, a nucleotide sequence encoding an antigen-binding molecule according to the present disclosure, e.g., a TCR or fragment thereof, is an mRNA.

[0530] In some embodiments, a nucleotide sequence encoding an antigen-binding molecule according to the present disclosure, e.g., a TCR or fragment thereof, is comprised / contained within a vector or plasmid. That is, the present disclosure provides a vector or plasmid comprising a nucleotide sequence encoding an antigen-binding molecule according to the present disclosure, e.g., a TCR or fragment thereof.

[0531] As referred to herein, a ‘vector’ may be a nucleic acid molecule used as a vehicle to transfer exogenous nucleic acid into a cell.

[0532] Accordingly, the present disclosure also provides a vector, or plurality of vectors, comprising the nucleic acid or plurality of nucleic acids according to the present disclosure. The vector may facilitate delivery of the nucleic acid(s) encoding a polypeptide according to the present disclosure to a cell. The vector may be an expression vector comprising elements required for expressing a polypeptide according to the present disclosure. The vector may comprise elements facilitating integration of the nucleic acid(s) into the genomic DNA of cell into which the vector is introduced.

[0533] A vector may be a vector for expression of the nucleic acid in a cell ( / .e., an expression vector). Vectors may include a promoter sequence operably linked to a nucleotide sequence encoding a TCR / antigen- binding molecule / polypeptide according to the present disclosure. A vector may also include a termination codon ( / .e., 3’ in the nucleotide sequence of the vector to the nucleotide sequence encoding the polypeptide(s)) and expression enhancers. Any suitable vectors, promoters, enhancers and termination codons known in the art may be used to express a peptide or polypeptide from a vector according to the present disclosure.

[0534] The term ‘operably linked’ may include the situation where nucleic acid encoding a polypeptide according to the present disclosure and regulatory nucleotide sequence(s) {e.g., a promoter and / or enhancers) are covalently linked in such a way as to place the expression of the nucleic acid encoding a polypeptide under the influence or control of the regulatory nucleotide sequence(s) (thereby forming an expression cassette). Thus, a regulatory sequence is operably linked to the selected nucleotide sequence if the regulatory sequence is capable of affecting transcription of the nucleotide sequence. The resulting transcript(s) may then be translated into the desired polypeptide(s). ‘Expression vector’ refers to a vector comprising a recombinant polynucleotide comprising expression control sequences operatively linked to a nucleotide sequence to be expressed. An expression vector comprises sufficient cis-acting elements for expression; other elements for expression can be supplied by the host cell or in an in vitro expression system. Expression vectors include all those known in the art, such as cosmids, plasmids (e.g., naked or contained in liposomes) and viruses (e.g., Sendai viruses, lentiviruses, retroviruses, adenoviruses, and adeno-associated viruses) that incorporate the recombinant polynucleotide. For example, the expression vector backbone may be the sequence of SEQ ID NO:171.

[0535] Vectors contemplated in connection with the present disclosure include DNA vectors, RNA vectors, plasmids (e.g., conjugative plasmids (e.g., F plasmids), non-conjugative plasmids, R plasmids, col plasmids, episomes), viral vectors (e.g., retroviral vectors, e.g., gammaretroviral vectors (e.g., murine Leukemia virus (MLV)-derived vectors, e.g., SFG vector), lentiviral vectors, adenovirus vectors, adeno- associated virus vectors, vaccinia virus vectors and herpesvirus vectors), transposon-based vectors, and artificial chromosomes (e.g., yeast artificial chromosomes), e.g., as described in Maus et al., Annu Rev Immunol (2014) 32:189-225 and Morgan and Boyerinas, Biomedicines (2016) 4:9, which are both hereby incorporated by reference in their entirety. In some embodiments, a vector according to the present disclosure is a lentiviral vector.

[0536] In some embodiments, a vector is selected based on tropism for a cell type / tissue / organ to which it is desired to deliver the nucleic acid. In some embodiments, a vector is selected based on tropism for a cell type in which it is desired to express the TCR / antigen-binding molecule / polypeptide(s). For example, it may be desired to deliver the nucleic acid / express the TCR / antigen-binding moleculeZpolypeptide(s) in an immune cell, e.g., a T cell.

[0537] In some embodiments, the nucleic acid is a vector suitable for delivering the nucleic acid encoding the antigen-binding molecule / TCR as a gene therapy. In some embodiments, the vector is an adeno- associated virus (AAV) vector. Adeno-associated virus vectors and their use to vector gene therapy is reviewed e.g., in Wang et al., Nat. Rev. Drug Discov. (2019) 18: 358-378 and Li and Samulski, Nat. Rev. Genet. (2020) 12: 255-272, both of which are hereby incorporated by reference in their entirety. In some embodiments, a vector may be an adeno-associated virus vector described in Wang et al., Nat. Rev. Drug Discov. (2019) 18: 358-378. In some embodiments, a vector may be an adeno-associated virus vector described in Li and Samulski, Nat. Rev. Genet. (2020) 12: 255-272.

[0538] In some embodiments, a vector may be an adeno-associated viral vector of one of the following serotypes: AAV1 , AAV2, AAV2i8, AAV5, AAV6, AAV8, AAV9, AAV9.45, AAV10 or AAVrh74.

[0539] In some embodiments, a vector may be a lentiviral vector. A ‘lentivirus’ as used herein refers to a genus of the Retroviridae family. Lentiviruses are unique among the retroviruses in being able to infect nondividing cells; they can deliver a significant amount of genetic information into the DNA of the host cell, so they are one of the most efficient methods of a gene delivery vector. HIV, SIV, and FIV are all examples of lentiviruses. Vectors derived from lentiviruses offer the means to achieve significant levels of gene transfer in vivo. For example, pALD lentiviral vectors can be obtained from Aldevron / Oxgene.

[0540] In some embodiments, the vector may be a eukaryotic vector, i.e., a vector comprising the elements necessary for expression of protein from the vector in a eukaryotic cell. In some embodiments, the vector may be a mammalian vector, e.g., comprising a cytomegalovirus (CMV) or SV40 promoter to drive protein expression.

[0541] In some embodiments a vector comprises modification to increase binding to and / or transduction of a cell-type of interest ( / .e., as compared to the level of binding / transduction by the unmodified vector). In some embodiments modification is to a capsid protein.

[0542] In some embodiments a vector comprises a capsid protein comprising a cell-targeting peptide. In some embodiments the cell-targeting peptide is a cell-targeting peptide described in Buning and Srivastava, Molecular Therapy: Methods & Clinical Development (2019) 12: 248-265, which is hereby incorporated by reference in its entirety.

[0543] In some embodiments a vector comprises a capsid protein comprising substitution to one or more tyrosine residues, e.g., one or more surface-exposed tyrosine residues. In some embodiments, one or more tyrosine residues of the capsid protein are substituted with phenylalanine. In some embodiments a vector comprises a capsid protein in which one or more tyrosine residues are substituted with another amino acid as described in lida et al., Biomed Res Int. (2013) 2013: 974819, which is hereby incorporated by reference in its entirety.

[0544] In some embodiments, a vector may be an adeno-associated virus vector described in Buning and Srivastava, supra. In some embodiments, a vector may be an adeno-associated virus vector described in lida et al., supra.

[0545] SEQ ID NO:170 herein provides an exemplary expression vector for the delivery and / or expression of TCR_A0427. In some embodiments, a vector is based on SEQ ID NO:170 or SEQ ID NO:171 herein. In some embodiments, any nucleic acid(s) / nucleotide sequence(s) encoding a TCR can be inserted into the backbone of SEQ ID NO:171.

[0546] In some embodiments the nucleic acid / vector comprises one or more sequences for controlling expression of the nucleic acid. Accordingly, in some embodiments the nucleic acid / vector comprises a control element for inducible expression of the nucleic acid.

[0547] A sequence for controlling expression of the nucleic acid may provide for expression of the nucleic acid by cells of a particular type or tissue. For example, expression may be under the control of a cell type- or tissue-specific promoter. The term ‘promoter’ as used herein is defined as a DNA sequence recognized by the synthetic machinery of the cell, or introduced synthetic machinery, required to initiate the specific transcription of a polynucleotide sequence.

[0548] Promoters for cell type- or tissue-specific expression of a nucleic acid in accordance with the present invention can be selected in accordance with the disease to be treated / prevented. For example, the promoter may drive expression in an immune cell. The promoter may be a tissue-specific promoter, e.g., which, when operably linked with a polynucleotide encodes or specified by a gene, causes the gene product to be produced in a cell substantially only if the cell is a cell of the tissue type corresponding to the promoter. Expression may be under the control of a constitutive promoter, e.g., a nucleotide sequence which, when operably linked with a polynucleotide which encodes or specifies a gene product, causes the gene product to be produced in a cell under most or all physiological conditions of the cell.

[0549] A sequence for controlling expression of the nucleic acid may provide for expression of the nucleic acid in response to e.g., a given agent / signal. For example, expression may be under the control of inducible promoter. The agent may provide for inducible expression of the nucleic acid in vivo by administration of the agent to a subject having been administered with a modified cell according to the disclosure, or ex vivo / in vitro by administration of the agent to cells in culture ex vivo or in vitro.

[0550] In some embodiments a nucleic acid or vector according to the present disclosure may employ a conditional expression system for controlling expression of the nucleic acid encoding the antigen-binding- molecule / TCR by cells comprising the nucleic acid / vector. ‘Conditional expression’ may also be referred to herein as ‘inducible expression’, and refers to expression contingent on certain conditions, e.g., the presence of a particular agent. Conditional expression systems are well known in the art and are reviewed e.g., in Ryding et al. Journal of Endocrinology (2001) 171 , 1-14, which is hereby incorporated by reference in its entirety.

[0551] Cells comprisinq / expressinq TCRs

[0552] The present disclosure provides a cell, or a plurality / population of cells, comprising / expressing antigenbinding molecules, e.g., T cell receptors (TCRs). The cell / cells may express or comprise an antigenbinding molecule, e.g., TCR or fragment / chain thereof, according to the present disclosure. The cell / cells may comprise or express nucleic acid encoding an antigen-binding molecule, e.g., TCR or fragment / chain thereof, according to the present disclosure. Such cells may be called TCR-expressing cells. It will be appreciated that a TCR-expressing cell comprises the TCR it expresses. It will also be appreciated that a cell expressing nucleic acid encoding a TCR also expresses and comprises the TCR encoded by the nucleic acid. The cell / cells may express or comprise an antigen-binding molecule, e.g., TCR or fragment / chain thereof, according to the present disclosure on the surface of the cell.

[0553] Aspects and embodiments of the present disclosure relate to host cells, and in particular immune cells. It will be appreciated that where cells are referred to herein in the singular ( / .e., ‘a / the cell’), a plurality / population of such cells are also contemplated.

[0554] In aspects and embodiments of the present disclosure, the cells are primary cells. That is, in some embodiments, the cells are / were isolated directly from living tissue / a living subject. The cells may be from any animal or human. The cells may be mammalian, more preferably human. The cells may be from a human patient.

[0555] In preferred embodiments, the host cell is an immune cell. An ‘immune cell’ may be a cell of hematopoietic origin, e.g., a neutrophil, eosinophil, basophil, dendritic cell, lymphocyte, or monocyte. A lymphocyte may be e.g., a T cell, B cell, NK cell, NKT cell or innate lymphoid cell (ILC), or a precursor thereof. The host cell / immune cell may express e.g., CD3 polypeptides (e.g., CD3y CD3e CD3 or CD36), TCR polypeptides (TCRa or TCRp), CD27, CD28, CD4 or CD8. In some embodiments, the host cell / immune cell is a T cell, e.g., a CD3+ T cell. In some embodiments, the T cell is a CD3+, CD4+ T cell. In some embodiments, the T cell is a CD3+, CD8+ T cell. In some embodiments, the T cell is a T helper cell (Th cell). In some embodiments, the T cell is a cytotoxic T cell (e.g., a cytotoxic T lymphocyte (CTL)).

[0556] An antigen-specific T cell may display certain functional properties of a T cell in response to the antigen / antigenic peptide for which the T cell is specific, or in response a cell comprising / expressing the antigen / antigenic peptide. In some embodiments, the properties are functional properties associated with effector T cells, e.g., cytotoxic T lymphocytes (CTLs).

[0557] In some embodiments, an antigen-specific T cell may display one or more of the following properties: cytotoxicity to a cell comprising / expressing the antigen / peptide thereof for which the T cell is specific; proliferation, IFNy expression, CD107a expression, IL-2 expression, TNFa expression, perforin expression, granzyme expression, granulysin expression, and / or FAS ligand (FASL) expression in response to stimulation with the antigen / peptide thereof for which the T cell is specific, or in response to exposure to a cell comprising / expressing the antigen / peptide thereof for which the T cell is specific.

[0558] Antigen-specific T cells according to the present disclosure express / comprise an antigen-binding molecule / TCR / fragment thereof that is capable of recognising a peptide of the antigen for which the T cell is specific when presented by the appropriate MHC molecule. In some embodiments, the antigen-specific immune cell is a T cell, e.g., a CD3+ T cell. In some embodiments, the T cell is a CD3+, CD4+ T cell. In some embodiments, the T cell is a CD3+, CD8+ T cell. In some embodiments, the T cell is a T helper cell (Th cell)). In some embodiments, the T cell is a cytotoxic T cell (e.g., a cytotoxic T lymphocyte (CTL)).

[0559] In some embodiments, an antigen-specific immune cell (e.g., an antigen-specific T cell) is specific for an antigen of WT-1. Such cells may be referred to as WT-1 -specific immune cells (e.g., WT-1 -specific T cells). A WT-1 -specific immune cell expresses / comprises a receptor (preferably a T cell receptor) capable of recognising a peptide of an antigen of WT-1 (e.g., when presented by an MHC molecule). In some embodiments, the WT-1 -specific immune cell expresses / comprises a TCR specific for a peptide of a WT- 1 antigen presented by MHC class I. In some embodiments, the WT-1 -specific immune cell expresses / comprises a TCR specific for SEQ ID NO:1 , 179, 180, 181 , or 182 presented by MHC class I.

[0560] In some embodiments, the WT-1 -specific immune cell expresses / comprises a TCR specific for VLDFAPPGA (SEQ ID NO:1) presented by MHC class I. In some embodiments, the WT-1-specific immune cell expresses / comprises a TCR specific for VLDFX1PPX2X3 (SEQ ID NO:179) presented by MHC class I. In some embodiments, the WT-1 -specific immune cell expresses / comprises a TCR specific for VLDFX1X2PX3X4 (SEQ ID NQ:180) presented by MHC class I. In some embodiments, the WT-1- specific immune cell expresses / comprises a TCR specific for VXiDFX2X3PX4X5 (SEQ ID NO:181) presented by MHC class I. In some embodiments, the WT-1-specific immune cell expresses / comprises a TCR specific for VLDFX1PX2GX3 (SEQ ID NO:182) presented by MHC class I.

[0561] Host cells, e.g., immune cells or T cells according to the present disclosure, may be described as ‘engineered cells’ or ‘genetically engineered cells’. As used herein, an ‘engineered cell’ refers to a cell, e.g., immune cell / T cell, that has been genetically modified as compared to a naturally-occurring cell. The term ‘genetically engineered’ or ‘engineered’ refers to a method of modifying the genome of a cell, including, but not limited to, deleting a coding or non-coding region or a portion thereof or inserting a coding region or a portion thereof. In some embodiments, the cell that is modified is a lymphocyte, e.g., a T cell or a modified cell that expresses CD3, which can either be obtained from a patient or a donor. The cell can be modified (e.g., as described herein) to express an exogenous construct, such as, e.g., an antigen-binding molecule / TCR disclosed herein, which can be incorporated into the cell's genome. In some embodiments, the cell is modified to express CD3.

[0562] In some embodiments, the engineered cells have an anti-cancer and / or anti-tumor effect.

[0563] Any host / immune / T cell provided herein may be provided in a ‘purified’, ‘substantially purified’ or ‘isolated’ form. As used herein, a ‘substantially purified’ cell is a cell that is essentially free of other cell types. A substantially purified cell also refers to a cell which has been separated from other cell types with which it is normally associated in its naturally occurring state. In some instances, a population of substantially purified cells refers to a homogenous or substantially homogenous population of cells. In other instances, this term refers simply to cells that have been separated from the cells with which they are naturally associated in their natural state, or have been separated / isolated / purified from naturally-occurring biological material. In some embodiments, the cells are cultured in vitro. In other embodiments, the cells are not cultured in vitro.

[0564] Cells described herein may comprise a combination of TCRs, e.g., one or more TCRs according to the present disclosure, and optionally one or more further TCRs. The TCRs may be introduced into the cells via the same nucleic acid / vector or via different nucleic acids / vectors.

[0565] A plurality / population of cells described herein may comprise two or more TCRs, including TCR(s) from the present disclosure. For example, cells within the plurality / population of cells may each comprise two or more TCRs. As another example, different cells within the plurality / population of cells may each comprise at least one TCR, where the TCRs are identical or non-identical. Different TCRs may be individually transduced into different host cells.

[0566] A host cell / immune cell comprising an antigen-binding molecule, e.g., a TCR, or a nucleic acid encoding an antigen-binding molecule, e.g., a TCR, according to the present disclosure may be characterised by reference to functional properties of the cell. In some embodiments a host cell / immune cell comprising an antigen-binding molecule, e.g., a TCR, or a nucleic acid encoding an antigen-binding molecule, e.g., a TCR, according to the present disclosure displays one or more of the following properties:

[0567] (a) expression of one or more cytotoxic / effector factors (e.g., IFNy, granzyme, perforin, granulysin, CD107a, TNFa, FASL) in response to cells presenting the MHC: peptide complex for which the TCR is specific;

[0568] (b) proliferation / population expansion, and / or growth factor (e.g., IL-2, GM-CSF) expression in response to cells presenting the MHC:peptide complex for which the TCR is specific;

[0569] (c) cytotoxicity to cells presenting the MHC:peptide complex for which the TCR is specific;

[0570] (d) no cytotoxicity (i.e., same as, substantially the same as, or below baseline) to cells which do not present the MHC:peptide complex for which the TCR is specific; and

[0571] (e) anti-cancer activity (e.g., cytotoxicity to cancer cells, tumor growth inhibition, reduction of metastasis, etc.) against cancer comprising cells presenting the MHC:peptide complex for which the TCR is specific. As used herein, ‘expression’ may be gene expression or protein expression. Gene expression encompasses transcription of DNA to RNA, and can be measured by various means known to those skilled in the art, for example by measuring levels of mRNA by quantitative real-time PCR (qRT-PCR), or by reporter-based methods. Similarly, protein expression can be measured by various methods well known in the art, e.g., by antibody-based methods, for example by western blot, immunohistochemistry, immunocytochemistry, flow cytometry, ELISA, ELISPOT, or reporter-based methods.

[0572] Cell proliferation / population expansion can be investigated by analysing cell division or the number of cells over a period of time. Cell division can be analysed, for example, by in vitro analysis of incorporation of3H-thymidine or by CFSE dilution assay, e.g., as described in Fulcher and Wong, Immunol Cell Biol (1999) 77(6): 559-564, hereby incorporated by reference in its entirety. Proliferating cells can also be identified by analysis of incorporation of 5-ethynyl-2'-deoxyuridine (Edll) by an appropriate assay, as described e.g., in Buck et al., Biotechniques. 2008 Jun; 44(7):927-9, and Sali and Mitchison, PNAS USA 2008 Feb 19; 105(7): 2415-2420, both hereby incorporated by reference in their entirety.

[0573] Cytotoxicity and cell killing can be investigated, for example, using any of the methods reviewed in Zaritskaya et al., Expert Rev Vaccines (2011), 9(6):601-616, hereby incorporated by reference in its entirety. Examples of in vitro assays of cytotoxicity / cell killing assays include release assays such as the51Cr release assay, the lactate dehydrogenase (LDH) release assay, the 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyl tetrazolium bromide (MTT) release assay, and the calcein-acetoxymethyl (calcein-AM) release assay. These assays measure cell killing based on the detection of factors released from lysed cells. Cell killing by a given cell type can be analysed e.g., by co-culturing the test cells with the given cell type, and measuring the number / proportion of cells viable / dead test cells after a suitable period of time. Other suitable assays include the xCELLigence real-time cytolytic in vitro potency assay described in Cerignoli et al., PLoS One. (2018) 13(3): e0193498 (hereby incorporated by reference in its entirety).

[0574] Cells may be evaluated for anti-cancer activity by analysis in an appropriate in vitro assay or in vivo model of the relevant cancer.

[0575] Methods for producing cells comprising / expressing an antigen-binding molecule / TCR of interest are well known to the skilled person, and generally comprise introducing nucleic acid(s) / vector(s) encoding constituent polypeptide(s) of the antigen-binding molecule / TCR into the cells.

[0576] Such methods may comprise nucleic acid transfer for permanent ( / .e., stable) or transient expression of the transferred nucleic acid. The transfer may involve methods of transfection, transformation or transduction. The term ‘transfected’ or ‘transformed’ or ‘transduced’ as used herein refers to a process by which exogenous nucleic acid is transferred or introduced into the host cell. A ‘transfected’ or ‘transformed’ or ‘transduced’ cell is one which has been transfected, transformed or transduced with exogenous nucleic acid. The cell includes the primary subject cell and its progeny. In some embodiments, following introduction into a cell, nucleic acid(s) encoding the polypeptide(s) of the TCR may be integrated into or form part of the genomic DNA of the cell. In some embodiments, following introduction into a cell nucleic acid(s) encoding the polypeptide(s) may be maintained extrachromosomally.

[0577] Any suitable genetic engineering platform may be used, and include gammaretroviral vectors, lentiviral vectors, adenovirus vectors, DNA transfection, transposon-based gene delivery and RNA transfection, for example as described in Maus et al., Annu Rev Immunol (2014) 32:189-225, hereby incorporated by reference in its entirety. Methods also include those described e.g., in Wang and Riviere Mol Ther Oncolytics. (2016) 3:16015, which is hereby incorporated by reference in its entirety. Suitable methods for introducing nucleic acid(s) / vector(s) into cells include transduction, transfection and electroporation.

[0578] Methods for generating / expanding populations of cells comprising / expressing the TCR in vitro / ex vivo are well known to the skilled person. Suitable culture conditions ( / .e., cell culture media, additives, stimulations, temperature, gaseous atmosphere), cell numbers, culture periods and methods for introducing nucleic acid(s) / vector(s) encoding polypeptide(s) of interest into cells, etc. can be determined by reference e.g., to WO 2018 / 177966 A1 . In some embodiments, a cell / population of cells according to the present disclosure is prepared under GMP (good manufacturing practice; e.g., as described in the guidelines for good manufacturing practice published by the European Commission (Volume 4 of ‘The rules governing medicinal products in the European Union’ contains guidance for the interpretation of the principles and guidelines of good manufacturing practices for medicinal products for human and veterinary use laid down in Commission Directives 91 / 356 / EEC, as amended by Directive 2003 / 94 / EC, and 91 / 412 / EEC respectively)) conditions.

[0579] Conveniently, cultures of cells according to the present disclosure may be maintained at 37°C in a humidified atmosphere containing 5% CO2. The cells of cell cultures can be established and / or maintained at any suitable density, as can readily be determined by the skilled person. Cultures can be performed in any vessel suitable for the volume of the culture, e.g., in wells of a cell culture plate, cell culture flasks, a bioreactor, etc. In some embodiments cells are cultured in a bioreactor, e.g., a bioreactor described in Somerville and Dudley, Oncoimmunology (2012) 1 (8): 1435-1437, which is hereby incorporated by reference in its entirety.

[0580] Introducing nucleic acid(s) into a cell may comprise transduction, e.g., lentiviral transduction. Transduction of immune cells with viral vectors is described e.g., in Simmons and Alberola-lla, Methods Mol Biol. (2016) 1323:99-108, which is hereby incorporated by reference in its entirety.

[0581] Introducing nucleic acid(s) into a cell comprise transfection, e.g., mRNA transfection. Transfection of immune cells with mRNA is described e.g., in Campillo-Davo et al., Pharmaceutics (2021) 13(3):396 and Chong et al., PeerJ (2021) 9:e11165 which are hereby incorporated by reference in their entirety.

[0582] Agents may be employed to enhance the efficiency of transduction / transfection. Hexadimethrine bromide (polybrene) is a cationic polymer which is commonly used to improve transduction, through neutralising charge repulsion between virions and sialic acid residues expressed on the cell surface. Other agents commonly used to enhance transduction / transfection include e.g., the poloxamer-based agents such as LentiBOOST (Sirion Biotech), Retronectin (Takara), Vectofusin (Miltenyi Biotech) and also SureENTRY (Qiagen) and ViraDuctin (Cell Biolabs). In some embodiments the methods comprise centrifuging the cells into which it is desired to introduce nucleic acid encoding polypeptide(s) of the TCR in the presence of cell culture medium comprising viral vector comprising the nucleic acid (referred to in the art as ‘spinfection’).

[0583] The methods generally comprise introducing a nucleic acid encoding polypeptide(s) of the TCR into a cell, and culturing the cell under conditions suitable for expression of the polypeptide(s) by the cell. In some embodiments, the methods comprise culturing immune cells into which nucleic acid encoding the polypeptide(s) has been introduced, in order to expand their number.

[0584] In some embodiments, the methods comprise analysing the cells to confirm successful introduction of the nucleic acid into the cells. In some embodiments, the methods comprise analysing the cells to confirm expression of the polypeptide(s) by the cells (e.g., via evaluation of a detectable entity).

[0585] In some embodiments the methods further comprise separating / isolating / purifying / enriching cells expressing the TCR e.g., from other cells (e.g., cells which do not express the TCR). Methods for purifying / isolating immune cells from heterogeneous populations of cells are well known in the art, and may employ e.g., fluorescence-activated cell sorting (FACS)- or magnetic-activated cell sorting (MACS)- based methods for sorting populations of cells based on the expression of the TCR / constituent polypeptide(s) thereof. In some embodiments, the methods comprise separating / isolating / purifying / enriching cells of a particular type, e.g., CD8+ T cells or CTLs expressing the TCR of interest.

[0586] Methods for producing cells according to the present disclosure may comprise modifying the cells to reduce the expression of a CD3-TCR complex polypeptide. In some embodiments, the methods comprise modifying nucleic acid (e.g., endogenous nucleic acid) encoding the CD3-TCR complex polypeptide.

[0587] Modification of a given target nucleic acid can be achieved in a variety of ways known to the skilled person, including modification of the target nucleic acid by homologous recombination, and target nucleic acid editing using site-specific nucleases (SSNs).

[0588] Suitable methods may employ targeting by homologous recombination, which is reviewed, for example, in Mortensen Curr Protoc Neurosci. (2007) Chapter 4:Unit 4.29 and Vasquez et al., PNAS 2001 , 98(15): 8403-8410, both of which are hereby incorporated by reference in their entirety. Targeting by homologous recombination involves the exchange of nucleotide sequence through crossover events guided by homologous sequences. Other suitable techniques include nucleic acid editing using SSNs. Gene editing using SSNs is reviewed e.g., in Eid and Mahfouz, Exp Mol Med. 2016 Oct; 48(10): e265, which is hereby incorporated by reference in its entirety. Enzymes capable of creating site-specific double strand breaks (DSBs) can be engineered to introduce DSBs to target nucleotide sequence(s) of interest. DSBs may be repaired by either error-prone non-homologous end-joining (NHEJ), in which the two ends of the break are rejoined, often with insertion or deletion of nucleotides. Alternatively, DSBs may be repaired by homology-directed repair (HDR), a high-fidelity mechanism in which a DNA template with ends homologous to the break site is supplied and introduced at the site of the DSB.

[0589] SSNs capable of being engineered to generate target nucleotide sequence-specific DSBs include zinc- finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced palindromic repeats / CRISPR-associated-9 (CRISPR / Cas9) systems. ZFN systems are reviewed e.g., in Umov et al., Nat Rev Genet. (2010) 11 (9):636-46, which is hereby incorporated by reference in its entirety. ZFNs comprise a programmable Zinc Finger DNA-binding domain and a DNA- cleaving domain (e.g., a Fokl endonuclease domain). The DNA-binding domain may be identified by screening a Zinc Finger array capable of binding to the target nucleotide sequence. TALEN systems are reviewed e.g., in Mahfouz et al., Plant Biotechnol J. (2014) 12(8):1006-14, which is hereby incorporated by reference in its entirety. TALENs comprise a programmable DNA-binding TALE domain and a DNA- cleaving domain {e.g., a Fokl endonuclease domain). TALEs comprise repeat domains consisting of repeats of 33-39 amino acids, which are identical except for two residues at positions 12 and 13 of each repeat which are repeat variable di-residues (RVDs). Each RVD determines binding of the repeat to a nucleotide in the target DNA sequence according to the following relationship: ‘HD’ binds to C, ‘NT binds to A, ‘NG’ binds to T and ‘NN’ or ‘NK’ binds to G (Moscou and Bogdanove, Science (2009)

[0590] 326(5959): 1501.). CRISPR / Cas9 and related systems e.g., CRISPR / Cpf1 , CRISPR / C2c1 , CRISPR / C2c2 and CRISPR / C2c3 are reviewed e.g., in Nakade et al., Bioengineered (2017) 8(3):265-273, which is hereby incorporated by reference in its entirety. These systems comprise an endonuclease {e.g., Cas9, Cpf1 etc.) and the single-guide RNA (sgRNA) molecule. The sgRNA can be engineered to target endonuclease activity to nucleotide sequences of interest.

[0591] In some embodiments, modifying nucleic acid {e.g., endogenous nucleic acid) encoding the CD3-TCR complex polypeptide in accordance with the present disclosure employs a site-specific nuclease (SSN) system targeting nucleic acid encoding the CD3-TCR complex polypeptide. The SSN system may be a ZFN system, a TALEN system, CRISPR / Cas9 system, a CRISPR / Cpf1 system, a CRISPR / C2c1 system, a CRISPR / C2c2 system or a CRISPR / C2c3 system.

[0592] In some embodiments, a method for producing a cell according to the present disclosure comprises introducing nucleic acid(s) encoding CRISPR / Cas9 system(s) targeting TRAC, TRBC1 and / or TRBC2 {e.g., TRAC and TRBC1) into a cell. In some embodiments, the nucleic acid(s) encode a CRISPR RNA (crRNA) targeting TRAC, TRBC1 and / or TRBC2 {e.g., TRAC and TRBC1 ; e.g., an exon of TRAC, TRBC1 and / or TRBC2 {e.g., TRAC and TRBC1)) and a trans-activating crRNA (tracrRNA) for processing the crRNA to its mature form.

[0593] Compositions

[0594] The present disclosure also provides compositions comprising the TCRs, antigen-binding molecules, polypeptides, nucleic acids, vectors and cells described herein.

[0595] The polypeptides, polypeptide complexes, nucleic acids, expression vectors and cells described herein may be formulated as pharmaceutical compositions or medicaments for clinical use, and may comprise a pharmaceutically-acceptable carrier, diluent, excipient or adjuvant. In preferred aspects and embodiments, the present disclosure provides a pharmaceutical composition or medicament comprising a cell according to the present disclosure. Thus, the present disclosure also provides a pharmaceutical composition / medicament comprising a polypeptide, polypeptide complex, nucleic acid / plurality, expression vector / plurality or cell / plurality described herein. In preferred embodiments, a pharmaceutical composition / medicament according to the present disclosure comprises a nucleic acid / plurality, expression vector / plurality or cell described herein.

[0596] The pharmaceutical compositions / medicaments of the present disclosure may comprise one or more pharmaceutically-acceptable carriers {e.g., liposomes, micelles, microspheres, nanoparticles), diluents / excipients {e.g., starch, cellulose, a cellulose derivative, a polyol, dextrose, maltodextrin, magnesium stearate), adjuvants, fillers, buffers, preservatives {e.g., vitamin A, vitamin E, vitamin C, retinyl palmitate, selenium, cysteine, methionine, citric acid, sodium citrate, methyl paraben, propyl paraben), anti-oxidants {e.g., vitamin A, vitamin E, vitamin C, retinyl palmitate, selenium), lubricants {e.g., magnesium stearate, talc, silica, stearic acid, vegetable stearin), binders {e.g., sucrose, lactose, starch, cellulose, gelatin, polyethylene glycol (PEG), polyvinylpyrrolidone (PVP), xylitol, sorbitol, mannitol), stabilisers, solubilisers, surfactants {e.g., wetting agents), masking agents or colouring agents {e.g., titanium oxide).

[0597] The term ‘pharmaceutically-acceptable’ as used herein pertains to compounds, ingredients, materials, compositions, dosage forms, etc., which are, within the scope of sound medical judgement, suitable for use in contact with the tissues of the subject in question {e.g., a human subject) without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. Each carrier, diluent, excipient, adjuvant, filler, buffer, preservative, anti-oxidant, lubricant, binder, stabiliser, solubiliser, surfactant, masking agent, colouring agent, flavouring agent or sweetening agent of a composition according to the present disclosure must also be ‘acceptable’ in the sense of being compatible with the other ingredients of the formulation. Suitable carriers, diluents, excipients, adjuvants, fillers, buffers, preservatives, anti-oxidants, lubricants, binders, stabilisers, solubilisers, surfactants, masking agents, colouring agents, flavouring agents or sweetening agents can be found in standard pharmaceutical texts, for example, Remington’s ‘The Science and Practice of Pharmacy’ (Ed. A. Adejare), 23rd Edition (2020), Academic Press.

[0598] The pharmaceutical compositions / medicaments according to the present disclosure may be formulated for administration to a subject, e.g., administration via a route of administration as appropriate for the nature of the composition / medicament and the disease / condition to be treated / prevented. Pharmaceutical compositions and medicaments of the present disclosure may be formulated for topical, parenteral, systemic, intracavitary, intravenous, intra-arterial, intramuscular, intrathecal, intraocular, intraconjunctival, intratumoral, subcutaneous, intradermal, intrathecal, oral or transdermal routes of administration. In some embodiments, a pharmaceutical composition / medicament may be formulated for administration by injection or infusion, or administration by ingestion.

[0599] Suitable formulations may comprise the cell provided in a sterile or isotonic medium. Medicaments and pharmaceutical compositions may be formulated in fluid, including gel form. Fluid formulations may be formulated for administration by injection or infusion {e.g., via catheter) to a selected region of the human or animal body.

[0600] In some embodiments, the pharmaceutical compositions / medicament is formulated for injection or infusion, e.g., into a blood vessel, tissue / organ of interest, or a tumor.

[0601] The present disclosure also provides methods for the production of pharmaceutically useful compositions, such methods of production may comprise one or more steps selected from: producing a cell or cells described herein; isolating / purifying a cell or cells described herein; and / or mixing a cell or cells described herein with a pharmaceutically-acceptable carrier, adjuvant, excipient or diluent. For example, a further aspect of the present disclosure relates to a method of formulating or producing a medicament or pharmaceutical composition for use in the treatment of a disease / condition (e.g., a disease / condition described herein), the method comprising formulating a pharmaceutical composition or medicament by mixing a cell described herein with a pharmaceutically-acceptable carrier, adjuvant, excipient or diluent.

[0602] Compositions described herein may comprise a combination of TCRs, e.g., TCRs according to the present disclosure, optionally in combination with other TCRs. The combination of TCRs may be provided as amino acid / nucleotide sequences, or may be provided in a cell / plurality of cells e.g., wherein each cell contains at least two different TCRs, or wherein at least two cells of the plurality each contain a different TCR.

[0603] Compositions according to the present disclosure may optionally be provided in isolated or purified form. For example, articles according to the present disclosure may be isolated / purified from naturally-occurring biological material.

[0604] Therapeutic and prophylactic applications

[0605] The TCRs, antigen-binding molecules, polypeptides, nucleic acids, vectors, cells and compositions described herein find use in therapeutic and prophylactic methods.

[0606] The present disclosure provides a TCR, antigen-binding molecule, polypeptide, nucleic acid (or plurality thereof), vector (or plurality thereof), cell (or plurality thereof), or composition described herein for use in a method of medical treatment or prophylaxis. Also provided is a TCR, antigen-binding molecule, polypeptide, nucleic acid (or plurality thereof), vector (or plurality thereof), cell (or plurality thereof), or composition described herein for use as a medicament. Also provided is a TCR, antigen-binding molecule, polypeptide, nucleic acid (or plurality thereof), vector (or plurality thereof), cell (or plurality thereof), or composition described herein for use in a method of treating or preventing a disease or condition described herein. Also provided is the use of a TCR, antigen-binding molecule, polypeptide, nucleic acid (or plurality thereof), vector (or plurality thereof), cell (or plurality thereof), or composition described herein in the manufacture of a medicament for treating or preventing a disease or condition described herein. Also provided is a method of treating or preventing a disease or condition described herein, comprising administering to a subject a therapeutically or prophylactically effective amount of a TCR, antigen-binding molecule, polypeptide, nucleic acid (or plurality thereof), vector (or plurality thereof), cell (or plurality thereof), or composition described herein.

[0607] The methods (of treatment) may be effective to reduce the development or progression of a disease / condition, alleviation of the symptoms of a disease / condition or reduction in the pathology of a disease / condition. The methods may be effective to prevent progression of the disease / condition, e.g., to prevent worsening of, or to slow the rate of development of, the disease / condition. In some embodiments, the methods may lead to an improvement in the disease / condition, e.g., a reduction in the symptoms of the disease / condition or reduction in some other correlate of the severity / activity of the disease / condition. In some embodiments, the methods may prevent development of the disease / condition to a later stage (e.g., a chronic stage or metastasis). In accordance with various aspects of the present disclosure, treatment or prevention of a disease / condition may comprise one or more of the following: reducing the number and / or activity of cells presenting the MHC:peptide complex for which the TCR is specific; cell killing of / cytotoxicity to cells presenting the MHC:peptide complex for which the TCR is specific; and anti-cancer activity {e.g., cytotoxicity to cancer cells, tumor growth inhibition, reduction of metastasis, etc.) against cancer cells presenting the MHC:peptide complex for which the TCR is specific.

[0608] It will be appreciated that articles of the present disclosure find use in the treatment / prevention of diseases / conditions that would derive therapeutic or prophylactic benefit from a reduction in the number or activity of cells expressing WT-1 -derived antigenic peptides, e.g., cells of a WT-1 -associated cancer.

[0609] For example, the disease / condition may be associated with expression of WT-1. The disease / condition may be described as a WT-1 -associated disease or condition.

[0610] For example, the disease / condition may be a disease / condition in which a cell comprising a WT-1 antigen or a cell comprising a peptide of a WT-1 antigen {e.g., a peptide of a WT-1 antigen described herein, e.g., SEQ ID NO:1 , 179, 180, 181 or 182) is pathologically implicated. Such diseases / conditions include those in which a cell comprising a WT-1 antigen or a cell comprising a peptide of a WT-1 antigen {e.g., a peptide of a WT-1 antigen described herein, e.g., SEQ ID NO:1 , 179, 180, 181 or 182) is positively- associated with the onset, development or progression of the disease / condition, and / or severity of one or more symptoms of the disease / condition, or in which such a cell is a risk factor for the onset, development or progression of the disease / condition.

[0611] In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is a disease / condition characterized by the presence of a cell comprising a WT-1 antigen or a cell comprising a peptide of a WT-1 antigen {e.g., SEQ ID NO:1 , 179, 180, 181 or 182). In some embodiments, the disease / condition is characterized by an increased number / proportion / activity of such cells as compared to the number / proportion / activity of such cells observed in the absence of the disease / condition {e.g., in a healthy subject, or in equivalent non-diseased tissue).

[0612] In some embodiments, the disease to be treated / prevented in accordance with the present disclosure is a cancer. Cancer may refer to any unwanted cell proliferation (or any disease manifesting itself by unwanted cell proliferation), neoplasm or tumor. The cancer may be benign or malignant and may be primary or secondary (metastatic). A ‘cancer’ or ‘cancer tissue’ can include a neoplasm or tumor. A neoplasm or tumor may be any abnormal growth or proliferation of cells and may be located in any tissue.

[0613] In some embodiments the cancer is a cancer in which WT-1 is pathologically implicated. The cancer may be a WT-1 -associated cancer.

[0614] The cancer may be of tissues / cells derived from e.g., the adrenal gland, adrenal medulla, anus, appendix, bladder, blood, bone, bone marrow, brain, breast, cecum, central nervous system (including or excluding the brain) cerebellum, cervix, colon, duodenum, endometrium, epithelial cells {e.g., renal epithelia), gallbladder, oesophagus, glial cells, heart, ileum, jejunum, kidney, lacrimal glad, larynx, liver, lung, lymph, lymph node, lymphoblast, maxilla, mediastinum, mesentery, myometrium, nasopharynx, omentum, oral cavity, ovary, pancreas, parotid gland, peripheral nervous system, peritoneum, pleura, prostate, salivary gland, sigmoid colon, skin, small intestine, soft tissues, spleen, stomach, testis, thymus, thyroid gland, tongue, tonsil, trachea, uterus, vulva, and / or white blood cells. Tumors may be nervous or non-nervous system tumors. Nervous system tumors may originate either in the central or peripheral nervous system, e.g., a glioma such as a glioblastoma (also called glioblastoma multiforme or “GBM”), medulloblastoma, meningioma, neurofibroma, ependymoma, Schwannoma, neurofibrosarcoma, astrocytoma and oligodendroglioma. Non-nervous system cancers / tumors may originate in any other non-nervous tissue, examples include melanoma, mesothelioma, lymphoma, myeloma, leukemia, Non-Hodgkin’s lymphoma (NHL), Hodgkin’s lymphoma, chronic myelogenous leukemia (CML), acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), cutaneous T cell lymphoma (CTCL), chronic lymphocytic leukemia (CLL), hepatoma, epidermoid carcinoma, prostate carcinoma, breast cancer, lung cancer, colon cancer, ovarian cancer, pancreatic cancer, thymic carcinoma, non-small cell lung carcinoma (NSCLC), hematologic cancer and sarcoma.

[0615] In some embodiments, the cancer is selected from the group consisting of: a solid cancer, a hematological cancer, gastric cancer (e.g., gastric carcinoma, gastric adenocarcinoma, gastrointestinal cancer, gastrointestinal adenocarcinoma), liver cancer (hepatocellular carcinoma, cholangiocarcinoma), head and neck cancer (e.g., head and neck squamous cell carcinoma (HNSCC)), oral cavity cancer (e.g., oropharyngeal cancer (e.g., oropharyngeal carcinoma), oral cancer, oral squamous cell carcinoma (OSCC), laryngeal cancer, nasopharyngeal carcinoma, oesophageal cancer), colorectal cancer (e.g., colorectal carcinoma), colon cancer, colon carcinoma, cervical carcinoma, prostate cancer, lung cancer (e.g., NSCLC, small cell lung cancer, lung adenocarcinoma, squamous lung cell carcinoma), bladder cancer, urothelial carcinoma (UC), urogenital cancer, skin cancer (e.g., melanoma, advanced melanoma), renal cell cancer (e.g., renal cell carcinoma), ovarian cancer (e.g., ovarian carcinoma), mesothelioma, breast cancer (e.g., TNBC), brain cancer (e.g., glioblastoma), prostate cancer, pancreatic cancer, a myeloid hematologic malignancy, a lymphoblastic hematologic malignancy, myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute lymphoblastic leukemia (ALL), lymphoma, non-Hodgkin’s lymphoma (NHL), thymoma or multiple myeloma (MM).

[0616] In some embodiments, the cancer is selected from a hematological cancer (e.g., leukemia, e.g., chronic myeloid leukemia (CML)), ovarian cancer, pancreatic cancer, colorectal cancer, gastric cancer, and brain cancer (e.g., glioblastoma). In some embodiments, the cancer is selected from a hematological cancer (e.g., leukemia, e.g., chronic myeloid leukemia (CML)), ovarian cancer, and brain cancer (e.g., glioblastoma). In some embodiments, the cancer is selected from a hematological cancer (e.g., leukemia, e.g., chronic myeloid leukemia (CML)), ovarian cancer, brain cancer (e.g., glioblastoma), and pancreatic cancer.

[0617] WT-1 , a cell comprising a WT-1 antigen or a cell comprising a peptide of a WT-1 antigen may be pathologically implicated in any of the cancers described herein. Any of the cancers herein may comprise a cell comprising / expressing a WT-1 antigen or a cell comprising / expressing a peptide of a WT-1 antigen (e.g., a peptide of a WT-1 antigen described herein, e.g., SEQ ID NO:1 , 179, 180, 181 or 182). Any of the cancers herein may express WT-1 . Any of the cancers herein may comprise a cell that expresses WT-1 .

[0618] Any of the cancers described herein may comprise a cell comprising / expressing a WT-1 antigen or a cell comprising / expressing a peptide of a WT-1 antigen (e.g., a peptide of a WT-1 antigen described herein, e.g., SEQ ID N0:1 , 179, 180, 181 or 182), and a major histocompatibility complex (MHC) molecule comprising a polypeptide (e.g., an MHC class I a chain polypeptide) encoded by, e.g., a HLA-A*02 allele, e.g., a HLA-A*02:01 allele, a HLA-A*02:07 allele, a HLA-A*02:02 allele. The peptide may be presented on the MHC molecule on the surface of the cell.

[0619] The cancer may be newly diagnosed, relapsed / refractory and / or metastatic. The cancer may be resistant to chemotherapy and / or radiotherapy.

[0620] In some embodiments, the cancer may be a relapsed cancer. As used herein, a ‘relapsed’ cancer refers to a cancer which responded to a treatment (e.g., chemotherapy or radiation therapy), but which has subsequently re-emerged / progressed, e.g., after a period of remission. For example, a relapsed cancer may be a cancer whose growth / progression was inhibited by a treatment (e.g., chemotherapy or radiation therapy), and which has subsequently grown / progressed. A cancer that is relapsed with respect to given treatment may be described as having acquired resistance to such treatment. A cancer that is relapsed may also be referred to as ‘recurrent’.

[0621] In some embodiments, the cancer may be a refractory cancer. As used herein, a ‘refractory’ cancer refers to a cancer which has not responded to a treatment (e.g., chemotherapy or radiation therapy). For example, a refractory cancer may be a cancer whose growth / progression was not inhibited by a treatment (e.g., chemotherapy or radiation therapy). In some embodiments a refractory cancer may be a cancer for which a subject receiving treatment for the cancer did not display a partial or complete response to the treatment. A cancer that is refractory with respect to given treatment may be described as having intrinsic resistance to such treatment.

[0622] In some embodiments, the cancer may be a metastatic cancer. As used herein, a ‘metastatic cancer’ refers to a cancer which has spread from its original (primary) location to other (secondary) parts of the body.

[0623] In some embodiments, a cancer may be a combination of the above cancers, For example, a metastatic cancer may be either newly diagnosed or relapsed (recurrent). As another example, a cancer that is relapsed (recurrent) and / or metastatic may also be refractory to one of more treatments.

[0624] Methods of the present invention can be used to reduce the size / volume of a tumor.

[0625] Administration of the antigen-binding molecules, TCRs, polypeptides, nucleic acids, vectors, cells and compositions of the present disclosure is preferably in a ‘therapeutically-effective’ or ‘prophylactically- effective’ amount, this being sufficient to show therapeutic or prophylactic benefit to the subject. The actual amount administered, and rate and time-course of administration, will depend on the nature and severity of the disease / condition and the particular article administered. Prescription of treatment, e.g., decisions on dosage etc., is within the responsibility of general practitioners and other medical doctors, and typically takes account of the disease / disorder to be treated, the condition of the individual subject, the site of delivery, the method of administration and other factors known to practitioners. The ability of a therapeutic agent to promote disease regression can be evaluated using a variety of methods known to the skilled practitioner, such as in human subjects during clinical trials, in animal model systems predictive of efficacy in humans, or by assaying the activity of the agent in in vitro assays. Examples of the techniques and protocols mentioned above can be found in Remington’s ‘The Science and Practice of Pharmacy’ (Ed. A. Adejare), 23rd Edition (2020), Academic Press.

[0626] ‘Administration’ refers to the physical introduction of an agent / article to a subject, using any of the various methods and delivery systems known to those skilled in the art.

[0627] Administration of the articles of the present disclosure may be e.g., parenteral, systemic, topical, intracavitary, intravascular, intravenous, intra-arterial, intramuscular, intrathecal, intraocular, intraconjunctival, intratumoral, subcutaneous, intradermal, oral or transdermal. Administration may be by injection, infusion or ingestion.

[0628] In some aspects and embodiments, articles of the present disclosure may be administered to a tissue / organ of interest (e.g., a tissue / organ affected by the disease / condition (e.g., a tissue / organ in which symptoms of the disease / condition manifest)). In some aspects and embodiments, articles of the present disclosure may be administered to the blood ( / .e., intravenous / intra-arterial administration) by injection or infusion (e.g., via cannula), or may be administered subcutaneously or orally.

[0629] Exemplary routes of administration for the articles disclosed herein include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal or other parenteral routes of administration, for example by injection or infusion. The phrase ‘parenteral administration’ as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal injection and infusion, as well as in vivo electroporation. ‘Parenteral’ administration of composition of the invention includes, e.g., subcutaneous (s.c.), intravenous (i.v.), intramuscular (i.m.), or intrasternal injection, or infusion techniques. In some embodiments, the formulation is administered via a non-parenteral route, e.g., orally. Other non- parenteral routes include a topical, epidermal or mucosal route of administration, for example, intranasally, vaginally, rectally, sublingually or topically. Administering can also be performed, for example, once, a plurality of times, and / or over one or more extended periods.

[0630] In some embodiments, therapeutic or prophylactic intervention according to the present disclosure may further comprise administering another agent for the treatment / prevention of the relevant disease / condition.

[0631] Administration of the TCRs, antigen-binding molecules, polypeptides, nucleic acids, vectors, cells and compositions described herein may be alone or in combination with other treatments, either simultaneously or sequentially dependent upon the condition to be treated.

[0632] In some embodiments, the TCRs, antigen-binding molecules, polypeptides, nucleic acids, vectors, cells and compositions described herein may be administered in combination with another TCR, antigenbinding molecule, polypeptide, nucleic acid, vector, cell or composition. In some embodiments, a subject is administered with a plurality of (e.g., 2, 3, 4, or more) non-identical TCRs, antigen-binding molecules, polypeptides, nucleic acids, vectors, cells and compositions, where for example at least one of the plurality is an article described herein. In some embodiments, a subject is administered with nucleic acidZvector(s) encoding a plurality of non-identical TCRs / antigen-binding molecules / polypeptides. In some embodiments, a subject is administered with cells comprising / expressing a plurality of non-identical TCRs / antigen-binding molecules / polypeptides, or cells comprising nucleic acidZvector(s) encoding a plurality of non-identical TCRs / antigen-binding molecules / polypeptides.

[0633] In some embodiments, a treatment or therapy described herein comprises administering at least one TCR, antigen-binding molecule, nucleic acid, vector, cell and / or composition according to the present disclosure in combination with at least one non-identical TCR, antigen-binding molecule, nucleic acid, vector, cell and / or composition. The at least one non-identical TCR, antigen-binding molecule, nucleic acid, vector, cell and / or composition may be one described herein. In some embodiments, a plurality of (e.g., 2, 3, 4, or more) non-identical TCRs, antigen-binding molecules, polypeptides, nucleic acids, vectors, cells and compositions comprises 1 , 2, 3, 4, 5, 6, or 7 non-identical TCRs, antigen-binding molecules, polypeptides, nucleic acids, vectors, cells or compositions described herein, optionally in combination with other TCRs, antigen-binding molecules, polypeptides, nucleic acids, vectors, cells or compositions that are not described herein.

[0634] Simultaneous administration refers to administration with another therapeutic agent together, for example as a pharmaceutical composition containing both agents (combined preparation), or immediately after each other and optionally via the same route of administration (e.g., to the same tissue, artery, vein or other blood vessel). Sequential administration refers to administration of one agent followed after a given time interval by separate administration of another agent. It is not required that the two agents are administered by the same route, although this is the case in some embodiments. The time interval may be any time interval.

[0635] Multiple doses of the polypeptides, nucleic acids, vectors, cells and compositions of the present disclosure may be provided. One or more, or each, of the doses may be accompanied by simultaneous or sequential administration of another therapeutic agent.

[0636] Multiple doses may be separated by a predetermined time interval, which may be selected to be one of 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 days, or 1 , 2, 3, 4, 5, or 6 months. By way of example, doses may be given once every 7, 14, 21 or 28 days (plus or minus 3, 2, or 1 days).

[0637] Administration may be alone or in combination with other treatments, either simultaneously or sequentially dependent upon the disease / condition to be treated. The TCR, antigen-binding molecule, nucleic acid, vector cell or composition described herein and another prophylactic / therapeutic agent may be administered simultaneously or sequentially.

[0638] In some embodiments, the methods comprise additional therapeutic or prophylactic intervention, e.g., for the treatment / prevention of a cancer. In some embodiments, the therapeutic or prophylactic intervention is selected from chemotherapy, immunotherapy, radiotherapy, surgery, vaccination and / or hormone therapy. In some embodiments, the therapeutic or prophylactic intervention comprises leukapheresis. In some embodiments, the therapeutic or prophylactic intervention comprises a stem cell transplant. In some embodiments, treatment of cancer further comprises chemotherapy and / or radiotherapy. Chemotherapy and radiotherapy respectively refer to treatment of a cancer with a drug or with ionising radiation (e.g., radiotherapy using X-rays or y-rays). The drug may be a chemical entity, e.g., small molecule pharmaceutical, antibiotic, DNA intercalator, protein inhibitor (e.g., kinase inhibitor), or a biological agent, e.g., antibody, antibody fragment, aptamer, nucleic acid (e.g., DNA, RNA), peptide, polypeptide, or protein. The drug may be formulated as a pharmaceutical composition or medicament. The formulation may comprise one or more drugs (e.g., one or more active agents) together with one or more pharmaceutically acceptable diluents, excipients or carriers.

[0639] Chemotherapy may involve administration of more than one drug. A drug may be administered alone or in combination with other treatments, either simultaneously or sequentially dependent upon the condition to be treated.

[0640] The chemotherapy may be administered by one or more routes of administration, e.g., parenteral, intravenous injection, oral, subcutaneous, intradermal or intratumoral.

[0641] The chemotherapy may be administered according to a treatment regime. The treatment regime may be a pre-determined timetable, plan, scheme or schedule of chemotherapy administration which may be prepared by a physician or medical practitioner and may be tailored to suit the patient requiring treatment. The treatment regime may indicate one or more of: the type of chemotherapy to administer to the patient; the dose of each drug or radiation; the time interval between administrations; the length of each treatment; the number and nature of any treatment holidays, if any etc. For a co-therapy a single treatment regime may be provided which indicates how each drug is to be administered.

[0642] Chemotherapeutic drugs may be selected from: Abemaciclib, Abiraterone Acetate, Abitrexate (Methotrexate), Abraxane (Paclitaxel Albumin-stabilized Nanoparticle Formulation), ABVD, ABVE, ABVE- PC, AC, Acalabrutinib, AC-T, Adcetris (Brentuximab Vedotin), ADE, Ado-Trastuzumab Emtansine, Adriamycin (Doxorubicin Hydrochloride), Afatinib Dimaleate, Afinitor (Everolimus), Akynzeo (Netupitant and Palonosetron Hydrochloride), Aldara (Imiquimod), Aldesleukin, Alecensa (Alectinib), Alectinib, Alemtuzumab, Alimta (Pemetrexed Disodium), Aliqopa (Copanlisib Hydrochloride), Alkeran for Injection (Melphalan Hydrochloride), Alkeran Tablets (Melphalan), Aloxi (Palonosetron Hydrochloride), Alunbrig (Brigatinib), Ambochlorin (Chlorambucil), Amboclorin (Chlorambucil), Amifostine, Aminolevulinic Acid, Anastrozole, Aprepitant, Aredia (Pamidronate Disodium), Arimidex (Anastrozole), Aromasin (Exemestane), Arranon (Nelarabine), Arsenic Trioxide, Arzerra (Ofatumumab), Asparaginase Erwinia chrysanthemi, Atezolizumab, Avastin (Bevacizumab), Avelumab, Axicabtagene Ciloleucel, Axitinib, Azacitidine, Bavencio (Avelumab), BEACOPP, Becenum (Carmustine), Beleodaq (Belinostat), Belinostat, Bendamustine Hydrochloride, BEP, Besponsa (Inotuzumab Ozogamicin) , Bevacizumab, Bexarotene, Bexxar (Tositumomab and Iodine I 131 Tositumomab), Bicalutamide, BiCNU (Carmustine), Bleomycin, Blinatumomab, Blincyto (Blinatumomab), Bortezomib, Bosulif (Bosutinib), Bosutinib, Brentuximab Vedotin, Brigatinib, BuMel, Busulfan, Busulfex (Busulfan), Cabazitaxel, Cabometyx (Cabozantinib-S- Malate), Cabozantinib-S-Malate, CAF, Calquence (Acalabrutinib), Campath (Alemtuzumab), Camptosar (Irinotecan Hydrochloride), Capecitabine, CAPOX, Carac (Fluorouracil-Topical), Carboplatin, CARBOPLATIN-TAXOL, Carfilzomib, Carmubris (Carmustine), Carmustine, Carmustine Implant, Casodex (Bicalutamide), CEM, Ceritinib, Cerubidine (Daunorubicin Hydrochloride), Cervarix (Recombinant HPV Bivalent Vaccine), Cetuximab, CEV, Chlorambucil, CHLORAMBUCIL-PREDNISONE, CHOP, Cisplatin, Cladribine, Clafen (Cyclophosphamide), Clofarabine, Clofarex (Clofarabine), Clolar (Clofarabine), CMF, Cobimetinib, Cometriq (Cabozantinib-S-Malate), Copanlisib Hydrochloride, COPDAC, COPP, COPP-ABV, Cosmegen (Dactinomycin), Cotellic (Cobimetinib), Crizotinib, CVP, Cyclophosphamide, Cyfos (Ifosfamide), Cyramza (Ramucirumab), Cytarabine, Cytarabine Liposome, Cytosar-U (Cytarabine), Cytoxan (Cyclophosphamide), Dabrafenib, Dacarbazine, Dacogen (Decitabine), Dactinomycin, Daratumumab, Darzalex (Daratumumab), Dasatinib, Daunorubicin Hydrochloride, Daunorubicin Hydrochloride and Cytarabine Liposome, Decitabine, Defibrotide Sodium, Defitelio (Defibrotide Sodium), Degarelix, Denileukin Diftitox, Denosumab, DepoCyt (Cytarabine Liposome), Dexamethasone, Dexrazoxane Hydrochloride, Dinutuximab, Docetaxel, Doxil (Doxorubicin Hydrochloride Liposome), Doxorubicin Hydrochloride, Doxorubicin Hydrochloride Liposome, Dox-SL (Doxorubicin Hydrochloride Liposome), DTIC-Dome (Dacarbazine), Durvalumab, Efudex (Fluorouracil-Topical), Elitek (Rasburicase), Ellence (Epirubicin Hydrochloride), Elotuzumab, Eloxatin (Oxaliplatin), Eltrombopag Olamine, Emend (Aprepitant), Empliciti (Elotuzumab), Enasidenib Mesylate, Enzalutamide, Epirubicin Hydrochloride, EPOCH, Erbitux (Cetuximab), Eribulin Mesylate, Erivedge (Vismodegib), Erlotinib Hydrochloride, Erwinaze (Asparaginase Erwinia chrysanthemi), Ethyol (Amifostine), Etopophos (Etoposide Phosphate), Etoposide, Etoposide Phosphate, Evacet (Doxorubicin Hydrochloride Liposome), Everolimus, Evista (Raloxifene Hydrochloride), Evomela (Melphalan Hydrochloride), Exemestane, 5-FU (Fluorouracil Injection), 5-FU (Fluorouracil-Topical), Fareston (Toremifene), Farydak (Panobinostat), Faslodex (Fulvestrant), FEC, Femara (Letrozole), Filgrastim, Fludara (Fludarabine Phosphate), Fludarabine Phosphate, Fluoroplex (Fluorouracil-Topical), Fluorouracil Injection, Fluorouracil-Topical, Flutamide, Folex (Methotrexate), Folex PFS (Methotrexate), FOLFIRI, FOLFIRI-BEVACIZUMAB, FOLFIRI-CETUXIMAB, FOLFIRINOX, FOLFOX, Folotyn (Pralatrexate), FU-LV, Fulvestrant, Gardasil (Recombinant HPV Quadrivalent Vaccine), Gardasil 9 (Recombinant HPV Nonavalent Vaccine), Gazyva (Obinutuzumab), Gefitinib, Gemcitabine Hydrochloride, GEMCITABINE-CISPLATIN, GEMCITABINEOXALIPLATIN, Gemtuzumab Ozogamicin, Gemzar (Gemcitabine Hydrochloride), Gilotrif (Afatinib Dimaleate), Gleevec (Imatinib Mesylate), Gliadel (Carmustine Implant), Gliadel wafer (Carmustine Implant), Glucarpidase, Goserelin Acetate, Halaven (Eribulin Mesylate), Hemangeol (Propranolol Hydrochloride), Herceptin (Trastuzumab), HPV Bivalent Vaccine, Recombinant, HPV Nonavalent Vaccine, Recombinant, HPV Quadrivalent Vaccine, Recombinant, Hycamtin (Topotecan Hydrochloride), Hydrea (Hydroxyurea), Hydroxyurea, Hyper-CVAD, Ibrance (Palbociclib), Ibritumomab Tiuxetan, Ibrutinib, ICE, Iclusig (Ponatinib Hydrochloride), Idamycin (Idarubicin Hydrochloride), Idarubicin Hydrochloride, Idelalisib, Idhifa (Enasidenib Mesylate), Ifex (Ifosfamide), Ifosfamide, Ifosfamidum (Ifosfamide), IL-2 (Aldesleukin), Imatinib Mesylate, Imbruvica (Ibrutinib), Imfinzi (Durvalumab), Imiquimod, Imlygic (Talimogene Laherparepvec), Inlyta (Axitinib), Inotuzumab Ozogamicin, Interferon Alfa-2b, Recombinant, lnterleukin-2 (Aldesleukin), Intron A (Recombinant Interferon Alfa-2b), Iodine I 131 Tositumomab and Tositumomab, Ipilimumab, Iressa (Gefitinib), Irinotecan Hydrochloride, Irinotecan Hydrochloride Liposome, Istodax (Romidepsin), Ixabepilone, Ixazomib Citrate, Ixempra (Ixabepilone), Jakafi (Ruxolitinib Phosphate), JEB, Jevtana (Cabazitaxel), Kadcyla (Ado-Trastuzumab Emtansine), Keoxifene (Raloxifene Hydrochloride), Kepivance (Palifermin), Keytruda (Pembrolizumab), Kisqali (Ribociclib), Kymriah (Tisagenlecleucel), Kyprolis (Carfilzomib), Lanreotide Acetate, Lapatinib Ditosylate, Lartruvo (Olaratumab), Lenalidomide, Lenvatinib Mesylate, Lenvima (Lenvatinib Mesylate), Letrozole, Leucovorin Calcium, Leukeran (Chlorambucil), Leuprolide Acetate, Leustatin (Cladribine), Levulan (Aminolevulinic Acid), Linfolizin (Chlorambucil), LipoDox (Doxorubicin Hydrochloride Liposome), Lomustine, Lonsurf (Trifluridine and Tipiracil Hydrochloride), Lupron (Leuprolide Acetate), Lupron Depot (Leuprolide Acetate), Lupron Depot-Ped (Leuprolide Acetate), Lynparza (Olaparib), Marqibo (Vincristine Sulfate Liposome), Matulane (Procarbazine Hydrochloride), Mechlorethamine Hydrochloride, Megestrol Acetate, Mekinist (Trametinib), Melphalan, Melphalan Hydrochloride, Mercaptopurine, Mesna, Mesnex (Mesna), Methazolastone (Temozolomide), Methotrexate, Methotrexate LPF (Methotrexate), Methylnaltrexone Bromide, Mexate (Methotrexate), Mexate-AQ (Methotrexate), Midostaurin, Mitomycin C, Mitoxantrone Hydrochloride, Mitozytrex (Mitomycin C), MOPP, Mozobil (Plerixafor), Mustargen (Mechlorethamine Hydrochloride), Mutamycin (Mitomycin C), Myleran (Busulfan), Mylosar (Azacitidine), Mylotarg (Gemtuzumab Ozogamicin), Nanoparticle Paclitaxel (Paclitaxel Albumin-stabilized Nanoparticle Formulation), Navelbine (Vi norelbine Tartrate), Necitumumab, Nelarabine, Neosar (Cyclophosphamide), Neratinib Maleate, Nerlynx (Neratinib Maleate), Netupitant and Palonosetron Hydrochloride, Neulasta (Pegfilgrastim), Neupogen (Filgrastim), Nexavar (Sorafenib Tosylate), Nilandron (Nilutamide), Nilotinib, Nilutamide, Ninlaro (Ixazomib Citrate), Niraparib Tosylate Monohydrate, Nivolumab, Nolvadex (Tamoxifen Citrate), Nplate (Romiplostim), Obinutuzumab, Odomzo (Sonidegib), OEPA, Ofatumumab, OFF, Olaparib, Olaratumab, Omacetaxine Mepesuccinate, Oncaspar (Pegaspargase), Ondansetron Hydrochloride, Onivyde (Irinotecan Hydrochloride Liposome), Ontak (Denileukin Diftitox), Opdivo (Nivolumab), OPPA, Osimertinib, Oxaliplatin, Paclitaxel, Paclitaxel Albumin-stabilized Nanoparticle Formulation, PAD, Palbociclib, Palifermin, Palonosetron Hydrochloride, Palonosetron Hydrochloride and Netupitant, Pamidronate Disodium, Panitumumab, Panobinostat, Paraplat (Carboplatin), Paraplatin (Carboplatin), Pazopanib Hydrochloride, PCV, PEB, Pegaspargase, Pegfilgrastim, Peginterferon Alfa-2b, PEG-lntron (Peginterferon Alfa-2b), Pembrolizumab, Pemetrexed Disodium, Perjeta (Pertuzumab), Pertuzumab, Platinol (Cisplatin), Platinol-AQ (Cisplatin), Plerixafor, Pomalidomide, Pomalyst (Pomalidomide), Ponatinib Hydrochloride, Portrazza (Necitumumab), Pralatrexate, Prednisone, Procarbazine Hydrochloride, Proleukin (Aldesleukin), Prolia (Denosumab), Promacta (Eltrombopag Olamine), Propranolol Hydrochloride, Provenge (Sipuleucel-T), Purinethol (Mercaptopurine), Purixan (Mercaptopurine), Radium 223 Dichloride, Raloxifene Hydrochloride, Ramucirumab, Rasburicase, R- CHOP, R-CVP, Recombinant Human Papillomavirus (HPV) Bivalent Vaccine, Recombinant Human Papillomavirus (HPV) Nonavalent Vaccine, Recombinant Human Papillomavirus (HPV) Quadrivalent Vaccine, Recombinant Interferon Alfa-2b, Regorafenib, Relistor (Methylnaltrexone Bromide), R-EPOCH, Revlimid (Lenalidomide), Rheumatrex (Methotrexate), Ribociclib, R-ICE, Rituxan (Rituximab), Rituxan Hycela (Rituximab and Hyaluronidase Human), Rituximab, Rituximab and Hyaluronidase Human, Rolapitant Hydrochloride, Romidepsin, Romiplostim, Rubidomycin (Daunorubicin Hydrochloride), Rubraca (Rucaparib Camsylate), Rucaparib Camsylate, Ruxolitinib Phosphate, Rydapt (Midostaurin), Sclerosol Intrapleural Aerosol (Talc), Siltuximab, Sipuleucel-T, Somatuline Depot (Lanreotide Acetate), Sonidegib, Sorafenib Tosylate, Sprycel (Dasatinib), STANFORD V, Sterile Talc Powder (Talc), Steritalc (Talc), Stivarga (Regorafenib), Sunitinib Malate, Sutent (Sunitinib Malate), Sylatron (Peginterferon Alfa- 2b), Sylvant (Siltuximab), Synribo (Omacetaxine Mepesuccinate), Tabloid (Thioguanine), TAC, Tafinlar (Dabrafenib), Tagrisso (Osimertinib), Talc, Talimogene Laherparepvec, Tamoxifen Citrate, Tarabine PFS (Cytarabine), Tarceva (Erlotinib Hydrochloride), Targretin (Bexarotene), Tasigna (Nilotinib), Taxol (Paclitaxel), Taxotere (Docetaxel), Tecentriq (Atezolizumab), Temodar (Temozolomide), Temozolomide, Temsirolimus, Thalidomide, Thalomid (Thalidomide), Thioguanine, Thiotepa, Tisagenlecleucel, Tolak (Fluorouracil-Topical), Topotecan Hydrochloride, Toremifene, Torisel (Temsirolimus), Tositumomab and Iodine I 131 Tositumomab, Totect (Dexrazoxane Hydrochloride), TPF, Trabectedin, Trametinib, Trastuzumab, Treanda (Bendamustine Hydrochloride), Trifluridine and Tipiracil Hydrochloride, Trisenox (Arsenic Trioxide), Tykerb (Lapatinib Ditosylate), Unituxin (Dinutuximab), Uridine Triacetate, VAC, Valrubicin, Valstar (Valrubicin), Vandetanib, VAMP, Varubi (Rolapitant Hydrochloride), Vectibix (Panitumumab), VelP, Velban (Vinblastine Sulfate), Velcade (Bortezomib), Velsar (Vinblastine Sulfate), Vemurafenib, Venclexta (Venetoclax), Venetoclax, Verzenio (Abemaciclib), Viadur (Leuprolide Acetate), Vidaza (Azacitidine), Vinblastine Sulfate, Vincasar PFS (Vincristine Sulfate), Vincristine Sulfate, Vincristine Sulfate Liposome, Vinorelbine Tartrate, VIP, Vismodegib, Vistogard (Uridine Triacetate), Voraxaze (Glucarpidase), Vorinostat, Votrient (Pazopanib Hydrochloride), Vyxeos (Daunorubicin Hydrochloride and Cytarabine Liposome), Wellcovorin (Leucovorin Calcium), Xalkori (Crizotinib), Xeloda (Capecitabine), XELIRI, XELOX, Xgeva (Denosumab), Xofigo (Radium 223 Dichloride), Xtandi (Enzalutamide), Yervoy (Ipilimumab), Yescarta (Axicabtagene Ciloleucel), Yondelis (Trabectedin), Zaltrap (Ziv-Aflibercept), Zarxio (Filgrastim), Zejula (Niraparib Tosylate Monohydrate), Zelboraf (Vemurafenib), Zevalin (Ibritumomab Tiuxetan), Zinecard (Dexrazoxane Hydrochloride), Ziv-Aflibercept, Zofran (Ondansetron Hydrochloride), Zoladex (Goserelin Acetate), Zoledronic Acid, Zolinza (Vorinostat), Zometa (Zoledronic Acid), Zydelig (Idelalisib), Zykadia (Ceritinib) and Zytiga (Abiraterone Acetate).

[0643] In some embodiments, the TCRs, antigen-binding molecules, nucleic acids, vectors, cells and compositions described herein are used in T-cell-based adoptive cell transfer (ACT), immunotherapy or cellular immunotherapy.

[0644] In some embodiments, the engineered TCRs described herein are exogenously expressed on T-cells through genetic engineering methods including, but not limited to, lentiviral transduction, or messenger ribonucleic acid (mRNA) transfection, of nucleic acids encoding for the TCR sequences described herein. In some embodiments, the TCRs used for ACT comprise a TCR sequence fused with a T-cell binding domain, including but not limited to a single-chain fragment binding to CD3. In some embodiments, the TCRs are used in T-cell based ACT in combination with one or more therapeutic agents, e.g., immune modulating agents, including but not limited to cytokines, TLR agonists, RIG-I like receptor (RLR) agonists.

[0645] Adoptive cell transfer (ACT) is an immunotherapy involving administration of immune cells with direct anticancer activity to a subject in need thereof. Adoptive cell transfer generally refers to a process by which cells (e.g., immune cells) are obtained from a subject, typically by drawing a blood sample from which the cells are isolated. The cells are then typically modified and / or expanded, and then administered either to the same subject (in the case of adoptive transfer of autologous / autogeneic cells) or to a different subject (in the case of adoptive transfer of allogeneic cells). The treatment is typically aimed at providing a population of cells with certain desired characteristics to a subject, or increasing the frequency of such cells with such characteristics in that subject. Adoptive transfer may be performed with the aim of introducing a cell or population of cells into a subject, and / or increasing the frequency of a cell or population of cells in a subject. The term ‘autologous’ refers to any material derived from the same individual to which it is later to be reintroduced. For example, an autologous T cell therapy comprises administering to a subject a T cell that was isolated from the same subject. The term ‘allogeneic’ refers to any material derived from one individual which is then introduced to another individual of the same species. For example, an allogeneic T cell transplantation comprises administering to a subject a T cell that was obtained from a donor other than the subject.

[0646] Adoptive transfer of immune cells is described, for example, in Kalos and June (2013), Immunity 39(1): 49-60, and Davis etal. (2015), Cancer J. 21 (6): 486-491 , both of which are hereby incorporated by reference in their entirety. The skilled person is able to determine appropriate reagents and procedures for adoptive transfer of cells according to the present disclosure, for example by reference to Dai etal., 2016 J Nat Cancer Inst 108(7): djv439, which is incorporated by reference in its entirety.

[0647] The term ‘immunotherapy’ or ‘cellular immunotherapy’ refers to the treatment of a subject afflicted with, or at risk of contracting or suffering a recurrence of, a disease by a method comprising inducing, enhancing, suppressing or otherwise modifying an immune response. Examples of immunotherapy include, but are not limited to, T cell therapies, antibody therapy, fusion protein therapy. T cell therapy can include adoptive T cell therapy, tumor-infiltrating lymphocyte (TIL) immunotherapy, autologous cell therapy, engineered autologous cell therapy (eACT), and allogeneic T cell transplantation (see, e.g., June, C. H., ed., 2001 , in: Cancer Chemotherapy and Biotherapy: Principles and Practice, Lippincott Williams & Wilkins, Baltimore; Vonderheide et al., 2003, Immun. Research 27: 1-15, which are hereby incorporated by reference in their entirety).

[0648] An ‘immune response’ refers to the action of a cell of the immune system (for example, T lymphocytes, B lymphocytes, natural killer (NK) cells, macrophages, eosinophils, mast cells, dendritic cells and neutrophils) and soluble macromolecules produced by any of these cells or the liver (including antibodies, cytokines, and complement) that results in selective targeting, binding to, damage to, destruction of, and / or elimination from a vertebrate's body of invading pathogens, cells or tissues infected with pathogens, cancerous or other abnormal cells, or, in cases of autoimmunity or pathological inflammation, normal human cells or tissues.

[0649] Cells used in immunotherapy described herein can come from any source known in the art. For example, T cells can be differentiated in vitro from a hematopoietic stem cell population, or T cells can be obtained from a subject. T cells can be obtained from, e.g., peripheral blood mononuclear cells, bone marrow, lymph node tissue, cord blood, thymus tissue, tissue from a site of infection, ascites, pleural effusion, spleen tissue, and tumors. In addition, the T cells can be derived from one or more T cell lines available in the art. T cells can also be obtained from a unit of blood collected from a subject using any number of techniques known to the skilled artisan, such as FICOLL.TM. separation and / or apheresis. Additional methods of isolating T cells for a T cell therapy are disclosed in U.S. Patent Publication No.

[0650] 2013 / 0287748, which is herein incorporated by reference in its entirety. An immunotherapy can also comprise administering a modified cell to a subject, wherein the modified cell expresses CD3 and a TCR disclosed herein. An immunotherapy can comprise administering a nucleic acid to a subject, e.g., using a vector or another type of targeting method, such that a cell is modified in vivo to express the nucleic acid. The nucleic acid can encode a TCR. In some embodiments, the modified cell is not a T cell. The advantages of ACT over other immunotherapies are multiple. Firstly, the antitumor T-cells can be grown in vitro in large numbers, then selected for high-avidity recognition of the desired tumor antigen, as well as effector functions. Secondly, in vitro activation circumvents the presence of inhibitory factors found in vivo. Thirdly, ACT allows for manipulation of the host before cell transfer to provide a favorable microenvironment supporting antitumor activity (Rosenberg and Restifo, Science 348(6230) :62-68 (2015)).

[0651] ACT using naturally occurring tumor-reactive T-cells achieves durable, complete regressions in patients with melanoma, as well as other common epithelial cancers (Rosenberg and Restifo, Science 348(6230) :62-68 (2015)). In some embodiments, TCR-expressing T-cells are used for T-cell-based adoptive cell transfer (ACT) as a therapeutic treatment in a subject suffering from cancer, including an EBV-associated cancer.

[0652] In other embodiments, ACT is used in combination with immune modulating agents, selected from the group of: immune cell-depleting agents, cytokines, TLR agonist, RIG-I like receptor (RLR) agonists, immune checkpoint inhibitors, chemotherapeutic agents, antibodies, radiotherapy and a combination thereof.

[0653] The present disclosure provides methods comprising administering antigen-specific immune cells comprising / expressing a TCR / antigen-binding molecule according to the present disclosure, or antigenspecific immune cells comprising / expressing nucleic acid / a vector encoding a TCR / antigen-binding molecule according to the present disclosure, to a subject.

[0654] In some embodiments, the methods comprise generating antigen-specific immune cells, or generating / expanding a population of antigen-specific immune cells. In some embodiments, the methods comprise modifying an immune cell to comprise / express a TCR / antigen-binding molecule according to the present disclosure. In some embodiments, the methods comprise modifying an immune cell to comprise / express nucleic acid / a vector encoding a TCR / antigen-binding molecule according to the present disclosure.

[0655] In some embodiments, the methods comprise administering to a subject antigen-specific immune cells modified to express / comprise a TCR / antigen-binding molecule according to the present disclosure (or modified to express / comprise a nucleic acid / vector encoding such a TCR / antigen-binding molecule).

[0656] In some embodiments, the methods comprise:

[0657] (a) modifying an immune cell to express or comprise a TCR / antigen-binding molecule according to the present disclosure, or to express or comprise nucleic acid / a vector encoding a TCR / antigen-binding molecule according to the present disclosure, and

[0658] (b) administering the immune cell modified to express or comprise a TCR / antigen-binding molecule according to the present disclosure, or modified to express or comprise nucleic acid / a vector encoding a TCR / antigen-binding molecule according to the present disclosure, to a subject.

[0659] In some embodiments, the methods comprise:

[0660] (a) isolating or obtaining immune cells; (b) modifying an immune cell to express or comprise a TCR / antigen-binding molecule according to the present disclosure, or to express or comprise nucleic acid / a vector encoding a TCR / antigen-binding molecule according to the present disclosure, and

[0661] (c) administering the immune cell modified to express or comprise a TCR / antigen-binding molecule according to the present disclosure, or modified to express or comprise nucleic acid / a vector encoding a TCR / antigen-binding molecule according to the present disclosure, to a subject.

[0662] In some embodiments, the subject from which the immune cells (e.g., PBMCs) are isolated is the same subject to which cells are administered ( / .e., adoptive transfer may be of autologous / autogeneic cells). In some embodiments, the subject from which the immune cells (e.g., PBMCs) are isolated is a different subject to the subject to which cells are administered ( / .e., adoptive transfer may be of allogeneic cells).

[0663] In some embodiments the methods may comprise one or more of: obtaining a blood sample from a subject; isolating immune cells (e.g., PBMCs) from a blood sample which has been obtained from a subject; culturing the immune cells in in vitro or ex vivo cell culture; modifying an immune cell to express or comprise a TCR / antigen-binding molecule according to the present disclosure, or to express or comprise nucleic acid / a vector encoding a TCR / antigen-binding molecule according to the present disclosure (e.g., by transduction with a viral vector encoding such TCR / antigen-binding molecule according to the present disclosure, or a viral vector comprising such nucleic acid); culturing immune cells expressing / comprising a TCR / antigen-binding molecule according to the present disclosure, or expressing / comprising a nucleic acid / a vector encoding a TCR / antigen-binding molecule according to the present disclosure in in vitro or ex vivo cell culture; collecting / isolating immune cells expressing / comprising a TCR / antigen-binding molecule according to the present disclosure, or expressing / comprising a nucleic acid / a vector encoding a TCR / antigen-binding molecule according to the present disclosure; formulating immune cells expressing / comprising a TCR / antigen-binding molecule according to the present disclosure, or expressing / comprising a nucleic acid / a vector encoding a TCR / antigen-binding molecule according to the present disclosure, to a pharmaceutical composition, e.g., by mixing the cells with a pharmaceutically acceptable adjuvant, diluent, or carrier; administering immune cells expressing / comprising a TCR / antigen-binding molecule according to the present disclosure, or expressing / comprising a nucleic acid / a vector encoding a TCR / antigen-binding molecule according to the present disclosure, or a pharmaceutical composition comprising such cells, to a subject.

[0664] In some embodiments, the methods may additionally comprise treating the cells or subject to induce / enhance expression of the TCR / antigen-binding molecule, and / or to induce / enhance proliferation or survival of immune cells comprising / expressing the TCR / antigen-binding molecule. In some embodiments, a subject is administered lymphodepleting chemotherapy prior to administration of immune cells expressing / comprising a TCR / antigen-binding molecule described herein (or expressing / comprising nucleic acid / a vector encoding such a TCR / antigen-binding molecule).

[0665] That is, in some embodiments, methods of treating / preventing a disease / condition in accordance with the present disclosure comprise: (i) administering a lymphodepleting chemotherapy to a subject, and (ii) subsequently administering an immune cell expressing / comprising a TCR / antigen-binding molecule described herein, or expressing / comprising nucleic acid / a vector encoding a TCR / antigen-binding molecule described herein.

[0666] As used herein, ‘lymphodepleting chemotherapy’ refers to treatment with a chemotherapeutic agent which results in depletion of lymphocytes (e.g., T cells, B cells, NK cells, NKT cells or innate lymphoid cell (ILCs), or precursors thereof) within the subject to which the treatment is administered. A ‘lymphodepleting chemotherapeutic agent’ refers to a chemotherapeutic agent which results in depletion of lymphocytes.

[0667] Lymphodepleting chemotherapy and its use in methods of treatment by adoptive cell transfer are described e.g., in Klebanoff et al., Trends Immunol. (2005) 26(2):111-7 and Muranski et al., Nat Clin Pract Oncol. (2006) (12):668-81 , both of which are hereby incorporated by reference in their entirety. The aim of lymphodepleting chemotherapy is to deplete the recipient subject’s endogenous lymphocyte population.

[0668] In the context of treatment of disease by adoptive transfer of immune cells, lymphodepleting chemotherapy is typically administered prior to adoptive cell transfer, to condition the recipient subject to receive the adoptively transferred cells. Lymphodepleting chemotherapy is thought to promote the persistence and activity of adoptively transferred cells by creating a permissive environment, e.g., through elimination of cells expressing immunosuppressive cytokines, and creating the ‘lymphoid space’ required for expansion and activity of adoptively transferred lymphoid cells.

[0669] Chemotherapeutic agents commonly used in lymphodepleting chemotherapy include e.g., fludarabine, cyclophosphamide, bedamustine and pentostatin.

[0670] In some embodiments, therapeutic or prophylactic intervention for the treatment / prevention of a disease / condition in accordance with the present disclosure comprises administration of a nucleic acid / vector, or of a composition comprising a nucleic acid / vector according to the present disclosure. In some embodiments, administration of such an article results in modification of a cell or cells (e.g., in vivo) to comprise / express a nucleic acid / vector, and / or to co mprise / ex press TCR / antigen-binding molecule / polypeptide(s) according to the present disclosure. That is, in some embodiments the nucleic acid / vector / composition is employed as a gene therapy.

[0671] In some aspects and embodiments in accordance with the present disclosure there may be targeted delivery of articles of the present disclosure, i.e., wherein the concentration of the relevant agent in the subject is increased in a given tissue(s) / organ(s) relative to other parts of the body. In some embodiments, the methods comprise intravascular (e.g., intravenous or intra-arterial), intramuscular or subcutaneous administration and wherein the relevant article is formulated in a targeted agent delivery system (e.g., as described herein). Suitable targeted delivery systems include, for example, nanoparticles, liposomes, micelles, beads, polymers, metal particles, dendrimers, antibodies, aptamers, nanotubes or micro-sized silica rods. Such systems may comprise a magnetic element to direct the agent to the desired organ or tissue. Suitable nanocarriers and delivery systems will be apparent to one skilled in the art.

[0672] The present disclosure further provides the use of an antigen-binding molecule / TCR / TCR fragment or chain / nucleic acid / vector / cell / composition according to the present disclosure to: recognize / bind to a cell expressing WT-1 (e.g., a WT-1 peptide, e.g., SEQ ID NO: 1 , 179, 180, 181 or 182, e.g., when presented by an appropriate MHC class I molecule); activate a host cell, e.g., T cell, in which the article is comprised / expressed; confer cytotoxic activity; kill cancer and / or tumor cells; kill cells expressing WT-1 ; and / or kill cancer / tumor cells expressing WT-1 .

[0673] The present disclosure further provides methods for recognizing / binding to a cell expressing WT-1 (e.g., a WT-1 peptide, e.g., SEQ ID NO:1 , 179, 180, 181 or 182, e.g., when presented by an appropriate MHC class I molecule); activating a host cell, e.g., T cell, in which the article is comprised / expressed; conferring cytotoxic activity; killing cancer and / or tumor cells; killing cells expressing WT-1 ; and / or killing cancer cells expressing WT-1 , using an antigen-binding molecule / TCR / TCR fragment or chain / nucleic acid / vector / cell / composition according to the present disclosure. Such uses / methods may be in vitro, or may be in vivo in a subject.

[0674] Accordingly, the present disclosure provides methods for recognizing / binding to a cell expressing WT-1 (e.g., a WT-1 peptide, e.g., SEQ ID NO:1 , 179, 180, 181 or 182, e.g., when presented by an appropriate MHC class I molecule); activating a host cell, e.g., T cell, in which the article is comprised / expressed; conferring cytotoxic activity; killing cancer and / or tumor cells; killing cells expressing WT-1 ; and / or killing cancer cells expressing WT-1 , comprising administering to a subject an antigen-binding molecule / TCR / TCR fragment or chain / nucleic acid / vector / cell / composition according to the present disclosure.

[0675] Subjects

[0676] A ‘subject’ as used herein includes any human who is afflicted with a disease / condition (e.g., a cancer) as described herein. The terms ‘subject’ and ‘patient’ are used interchangeably herein.

[0677] The subject in accordance with aspects of the present disclosure may be any animal or human. The subject is preferably mammalian, more preferably human. The subject may be a non-human mammal, but is more preferably human. The subject may be male or female. The subject may be a patient. A subject may have been diagnosed with a disease or condition described herein requiring treatment (e.g., a cancer), may be suspected of having such a disease / condition, or may be at risk of developing / contracting such a disease / condition.

[0678] In embodiments according to the present disclosure, the subject is preferably a human subject. In some embodiments, the subject to be treated according to a therapeutic or prophylactic method of the present disclosure is a subject having, or at risk of developing, a disease / condition described herein e.g., a cancer, e.g., a cancer described herein. In embodiments according to the present disclosure, a subject may be selected for treatment according to the methods based on characterisation for certain markers of such a disease / condition. In some embodiments, a subject may comprise cells presenting a peptide:MHC complex described herein. In some embodiments, a subject may be selected for treatment described herein based on the detection of a cancer comprising / expressing WT-1 , e.g., in a sample obtained from the subject {e.g., a biopsy, e.g., of a tumor). In some embodiments, a subject may comprise cells comprising / expressing / overexpressing WT-1. In some embodiments, a subject may comprise cells comprising / expressing / overexpressing a peptide described herein, e.g., SEQ ID NO:1 , 179, 180, 181 or 182.

[0679] In some embodiments, a subject comprises an HLA allele as described herein. In some embodiments, a subject comprises an HLA-A*02 allele. In some embodiments, a subject comprises HLA-A*02:01 . In some embodiments, a subject comprises 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*02:11 , HLA-A*02:12, HLA-A*02:19, HLA-A*02:24, HLA-A*02:264, or HLA- A*02:52.

[0680] A subject to be administered immune cells in accordance with the present disclosure may be autogeneic / autologous with respect to the subject from which immune cells administered to the subject are derived. A subject to be administered immune cells in accordance with the present disclosure may be genetically identical to the subject from which immune cells administered to the subject are derived. A subject to be administered immune cells in accordance with the present disclosure may be the same subject as the subject from which immune cells administered to the subject are derived. A subject to be treated / prevented in accordance with the present disclosure may be HLA-matched with respect to the subject from which immune cells administered to the subject are derived. A subject to which cells are administered may comprise MHC / HLA genes encoding MHC / HLA molecules which are identical to the MHC / HLA molecules encoded by the MHC / HLA genes of the subject from which immune cells administered to the subject are derived.

[0681] A subject to be administered immune cells in accordance with the present disclosure may be allogeneic / non-autologous with respect to the subject from which immune cells administered to the subject are derived. A subject to be administered immune cells in accordance with the present disclosure may be genetically non-identical to the subject from which immune cells administered to the subject are derived. A subject to be administered immune cells in accordance with the present disclosure may be a different subject to the subject from which immune cells administered to the subject are derived. A subject to be treated / prevented in accordance with the present disclosure may be HLA-mismatched with respect to the subject from which immune cells administered to the subject are derived. A subject to which cells are administered may comprise MHC / HLA genes encoding MHC / HLA molecules which are non-identical to the MHC / HLA molecules encoded by the MHC / HLA genes of the subject from which immune cells administered to the subject are derived.

[0682] In some embodiments, the subject is a >4 / 8 ( / .e., 4 / 8, 5 / 8, 6 / 8, 7 / 8 or 8 / 8) match across HLA-A, -B, -C, and -DRB1. In some embodiments, the subject is a >5 / 10 ( / .e., 5 / 10, 6 / 10, 7 / 10, 8 / 10, 9 / 10 or 10 / 10) match across HLA-A, -B, -C, -DRB1 and -DQB1. In some embodiments, the subject is a >6 / 12 ( / .e., 6 / 12, 7 / 12 8 / 12, 9 / 12, 10 / 12, 11 / 12 or 12 / 12) match across HLA-A, -B, -C, -DRB1 , -DQB1 and -DPB1. In some embodiments, the subject is an 8 / 8 match across HLA-A, -B, -C, and -DRB1 . In some embodiments, the subject is a 10 / 10 match across HLA-A, -B, -C, -DRB1 and -DQB1. In some embodiments, the subject is a 12 / 12 match across HLA-A, -B, -C, -DRB1 , -DQB1 and -DPB1.

[0683] Kits

[0684] The present disclosure also provides kits of parts. Aspects and embodiments of the present disclosure relate to kits for producing a cell {e.g., an antigen-specific cell) according to the present disclosure.

[0685] Aspects and embodiments of the present disclosure relate to kits for performing the methods according to the present disclosure, in whole or in part.

[0686] In some embodiments, the kit may have at least one container having a predetermined quantity of a TCR, antigen-binding molecule, polypeptide, nucleic acid, vector, cell or composition described herein. The kit may provide the relevant articles together with instructions {e.g., a protocol) as to how to employ them in accordance with a method described herein.

[0687] In some embodiments, a kit of parts comprises materials for producing a polypeptide according to the present disclosure. In some embodiments, a kit of parts comprises materials for producing a TCR / antigen-binding molecule according to the present disclosure. In some embodiments, a kit of parts comprises materials for producing a cell according to the present disclosure. In some embodiments, a kit of parts comprises materials for producing a composition according to the present disclosure.

[0688] In some embodiments, the kit of parts may comprise a nucleic acid / plurality or an expression vector / plurality according to the present disclosure, and optionally materials for introducing the nucleic acid / plurality or an expression vector / plurality into a cell.

[0689] In some embodiments, the kit may comprise materials for producing a TCR, antigen-binding molecule, polypeptide, nucleic acid, vector, cell or composition described herein. In some embodiments, the kit of parts may comprise materials for formulating a TCR, antigen-binding molecule, polypeptide, nucleic acid, vector, cell or composition described herein to a pharmaceutical composition / medicament, e.g., in a composition further comprising a pharmaceutically-acceptable carrier, diluent, excipient or adjuvant.

[0690] The kit may provide a TCR, antigen-binding molecule, polypeptide, nucleic acid, vector, cell or composition described herein together with instructions for administration to a patient in order to treat a specified disease / condition {e.g., a disease / condition described herein).

[0691] In some embodiments the kit may further comprise at least one container having a predetermined quantity of another therapeutic agent {e.g., as described herein). In such embodiments, the kit may also comprise a second medicament or pharmaceutical composition such that the two medicaments or pharmaceutical compositions may be administered simultaneously or separately such that they provide a combined treatment for the specific disease / condition.

[0692] Kits according to the present disclosure may include instructions for use, e.g., in the form of an instruction booklet or leaflet. The instructions may include a protocol for performing any one or more of the methods described herein. Sequence identity

[0693] As used herein, ‘sequence identity’ refers to the percent of nucleotides / amino acid residues in a subject sequence that are identical to nucleotides / amino acid residues in a reference sequence, after aligning the sequences and, if necessary, introducing gaps, to achieve the maximum percent sequence identity

[0694] 5 between the sequences. Pairwise and multiple sequence alignment for the purposes of determining percent sequence identity between two or more amino acid or nucleic acid sequences can be achieved in various ways known to a person of skill in the art, for instance, using publicly available computer software such as ClustalOmega (Soding, J. 2005, Bioinformatics 21 , 951-960), T-coffee (Notredame et al. 2000, J. Mol. Biol. (2000) 302, 205-217), Kalign (Lassmann and Sonnhammer 2005, BMC Bioinformatics, 6(298))

[0695] 10 and MAFFT (Katoh and Standley 2013, Molecular Biology and Evolution, 30(4) 772-780) software. When using such software, the default parameters, e.g., for gap penalty and extension penalty, are preferably used. Unless stated otherwise, all references herein to a percent sequence identity refer to percent sequence identity determined using ClustalOmega (Soding, J. 2005, Bioinformatics 21 , 951-960), using the default parameters.

[0696] 15 Sequences

[0697]

[0698]

[0699]

[0700] Table A. Amino acid sequences of the TCR CDR sequences

[0701] Table B. Amino acid sequences of the TCR FR sequences

[0702] Table C. Amino acid sequences of the TCRa and TCRP variable domains

[0703] Table D. Nucleotide of the TCRa and TCRB variable domains ***

[0704] The present disclosure includes the combination of the aspects and preferred features described except where such a combination is clearly impermissible or expressly avoided.

[0705] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0706] Aspects and embodiments of the present disclosure will now be illustrated, by way of example, with reference to the accompanying figures. Further aspects and embodiments will be apparent to those skilled in the art. All documents mentioned in this text are incorporated herein by reference.

[0707] Throughout this specification, including the claims which follow, unless the context requires otherwise, the word ‘comprise,’ and variations such as ‘comprises’ and ‘comprising,’ will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

[0708] It must be noted that, as used in the specification and the appended claims, the singular forms ‘a’, ‘an’, and ‘the’ include plural referents unless the context clearly dictates otherwise. Ranges may be expressed herein as from ‘about’ one particular value, and / or to ‘about’ another particular value. When such a range is expressed, another embodiment includes from the one particular value and / or to the other particular value. Similarly, when values are expressed as approximations, by the use of the antecedent ‘about’, it will be understood that the particular value forms another embodiment.

[0709] Where a nucleic acid sequence is disclosed or referred to herein, the reverse complement thereof is also expressly contemplated. Methods described herein may preferably be performed in vitro. The term ‘in vitro’ is intended to encompass procedures performed outside of a body such as a human body, e.g., with cells in culture, whereas the term ‘in vivo’ is intended to encompass procedures with / on intact multi-cellular organisms, e.g., within a body such as a human body.

[0710] Values may be expressed herein as ‘about’ a particular value. Similarly, ranges may be expressed herein as from ‘about’ a particular value, and / or to ‘about’ another particular value. The term ‘about’ in relation to a numerical value is optional, and means for example + / - 10 %. By way of illustration, reference e.g., to ‘about 10 %’ is to be construed as 9 % to 11 %. In instances herein where ‘about’ is recited, the value it precedes is also specifically contemplated. By way of illustration, reference e.g., to ‘about 10 %’ also specifically contemplates 10 %.

[0711] Unless indicated otherwise, the term ‘substantially’ is intended to encompass both ‘wholly’ and ‘largely but not wholly’. For example, a ‘substantially homogenous population of cells’ means a population of cells that is wholly ( / .e., entirely) homogenous, or largely but not wholly homogenous.

[0712] Brief Description of the Figures

[0713] Embodiments and experiments illustrating the principles of the invention will now be discussed with reference to the accompanying figures.

[0714] Figures 1A-1 E. Functional validation of WT-1 specific TCRs. (1A, 1 B) Graphs showing functional activation of Jurkat reporter cells transduced with WT-1 specific TCRs and exposed to target cells pulsed with a range of concentrations of WT-137-45 (SEQ ID NO:1). Estimated EC50 values are shown. (1C, 1 D) Graphs showing functional activation of Jurkat reporter cells electroporated with mRNA for WT-1 specific TCRs and exposed to target cells pulsed with a range of concentrations of WT-137-45 (SEQ ID NO:1). Estimated EC50 values are shown. (1 E) Graph showing T cell activation as measured through levels of IFNy secretion. Primary T cells transduced with WT-1 specific TCRs were exposed to target cells pulsed with a range of concentrations of WT-137-45 (SEQ ID NO:1). Culture supernatants were then analysed for levels of IFNy with non-transduced T cells from the same donor serving as baseline adjustment.

[0715] Figures 2A-2F. Cytotoxic activity of primary T cells transduced with WT-1 specific TCRs. (2A)

[0716] Graphs showing direct cytotoxic activity of primary T cells transduced with WT-1 specific TCRs as measured via flow cytometric quantification of CTV-stained target cells pulsed with a range of concentrations of WT-137-45 (SEQ ID NO:1). Experiments were carried out at an E:T ratio of 1 :1 with nontransduced T cells from the same donor indicating background levels of cytotoxicity. (2B, 2C, 2D) Graphs showing IFNy secretion as a measure of cytotoxic activity of primary T cells transduced with TCR_A0431 against endogenous levels of WT-137-45 (SEQ ID NO:1) in CML cell line (K562 + HLA-A*02:01), ovarian cancer cell line (COV434 + HLA-A*02:01) and glioblastoma cell line (SF539). Experiments were carried out at 3 different E:T ratios of 0.3:1 , 1 :1 and 3:1 with means and standard deviations shown from duplicate measurements, except for single-measurements in non-transduced (NT) cell controls. ****p < 0.0001 ; two-way ANOVA followed by Sidak’s multiple comparisons test. (2E, 2F) Graphs showing results for xCELLigence technology as used to measure specific killing of cancer cell lines expressing endogenous levels of WT-137-45 and HLA-A*02:01 by primary T cells transduced with TCR_A0427 or TCR_A0431 over a period of 150 hours. Data shown are for two E:T ratio, 1 :1 and 3:1 , in the context of a ovarian cancer cell line (COV434 + HLA-A*02:01) or glioblastoma cell line (SF539). Non-transduced primary T cells from the same donor serve as negative control while Tween-20 treatment was a positive control for complete cell lysis.

[0717] Figures 3A-3G. Alanine scan and safety assessment for potential binding of TCRs to endogenous proteins. (3A, 3B, 3C, 3D) Graphs showing alanine scan of TCR_A0427, TCR_A0429, TCR_A0431 and TCR_A0432 for target peptide, WT-I37-45 (SEQ ID NO:1). Raji cells expressing HLA-A*02:01 were pulsed with EC90 levels of WT-I37-45 (SEQ ID NO:1) carrying single alanine-substitutions at indicated positions (SEQ ID NO:172, 173, 174, 175, 176, 177 and 178). Primary T cells transduced with WT-1 specific TCRs were introduced at a 1 :1 ratio and T cell activation was measured through levels of IFNy secretion. Means and standard deviations from duplicate measurements were shown with values adjusted against nopeptide, non-transduced controls. (3E, 3F, 3G) Graphs showing potential alloreactivity of WT-1 specific TCRs, TCR_A0427 and TCR_A0431 , was assessed against a panel of 35 LCL lines representing 73 unique HLA alleles, including 10 of the most frequent HLA-A,B,C in North America, Europe, and Northeast Asia. Each TCR was evaluated in two different primary T cell donors (donor A or B) where T cell activation was measured through levels of IFNy secretion. Means and standard deviations from duplicate measurements were shown with values adjusted against non-transduced T cell controls.

[0718] Figures 4A-4E. WT-1 specific TCR functionally recognizes the target peptide, WT-137-45, presented on HLA-A*02 alleles beyond A*02:01. (4A) Dose-dependent activation of Jurkat-TCR A0431 by peptide-pulsed HLA-A*02 LCLs. Graphs showing functional activation of Jurkat reporter cells transduced with WT-1 specific TCRs and exposed to LCLs pulsed with a range of concentrations of WT-137-45 (SEQ ID NO:1). LCL homozygous for A*02:01 was used as a positive control while a LCL without any A*02 allele served as the negative control. Two different donor lines heterozygous for A*02:06, one for A*02:07 and one for A*02:02 were tested. Data shown are means from 2 replicate wells with error bars representing standard deviations. (4B) Bar chart showing the specific killing of Panel , Panel + HLA- A*02:01 or Panel + HLA-A*02:07 cell lines by primary T cells transduced with TCR_A0431 or nontransduced (NT) after 24h of co-incubation with target cells at E:T 3:1 . Data shown are cytolytic activities measured using real-time cell analysis (RTCA, xCELLigence) with mean and standard deviations from three different donors (donor A, B and C). (4C, 4D, 4E) Real-time cell analysis data, up to 24h, of TCR_A0431 transduced or non-transduced primary T cells from 3 different donors against Panel , Panel + HLA-A*02:01 or Panel + HLA-A*02:07 target cells.

[0719] Examples

[0720] Example 1 : Functional validation of WT-1 specific T cell receptors (TCRs)

[0721] 1.1. Materials and Methods

[0722] In vitro expansion of WT-1 specific T cells. CD14+ cells were first isolated from healthy donors’ peripheral blood mononuclear cells (PBMCs) before overnight culture in AIM-V with 2% AB serum, GM- CSF and IL-4 to facilitate their differentiation into activated monocytes (acMos). Simultaneously, the CD14- fractions were depleted of CD4+ and NKp46+ cell populations to generate a CD8+ enriched population that was then stimulated overnight with anti-CD28 antibody and target peptide, WT-137-45.

[0723] The next day, acMos were driven towards maturation for 2hrs with the addition of target peptide, R848, polyinosinic-polycytidylic acid (poly l:C) and dendritic cell maturation mix (Stemcell). Concurrently, previously stimulated, donor-matched CD8+ enriched populations were purified for CD137+ cells. These were then mixed with their respective, matured acMos at a 1 :1 ratio in AIM-V media with 2% AB serum, IL-7, IL-15 and IL-21 for 10 days. After 10 days of expansion, cultures were checked for WT-1 specific T cells through staining with WT-137-45 MHC tetramers before analysis via flow cytometry. A second stimulation with Transact beads was then performed and cells were expanded for another 10 days to increase the number of WT-1 specific T cells for effective TCR sequence recovery.

[0724] TCR sequence recovery. In vitro expanded T cells were stained with WT-I37-45 MHC tetramers and surface antibodies to sort specific T cells into individual wells for single cell PCR (scPCR). A Melody flow cytometer (Becton Dickinson BD) was used for the sorting. Sorted cells were immediately frozen and stored at -80C until further use. RT reaction was performed, followed by a nested PCR protocol to amplify the variable TCR regions of the TCR alpha (TCRa) and TCR beta (TCRp) chains using primers binding in the V gene. The protocol was described previously in Albert C. Shaw (ed.), Immunosenescence: Methods and Protocols, Methods in Molecular Biology, vol. 1343, which is hereby incorporated herein by reference in its entirety. TCR sequences were obtained by Sanger sequences of PCR products.

[0725] Plasmids, Lentiviral vector or mRNA production and transduction of Jurkat cells. Lentiviral vectors were produced for each TCR by transfection of the transgene plasmid along with packaging plasmids encoding the additional lentiviral components into HEK293 cells. In some examples, TCR sequences were codon optimised with an algorithm provided by Genscript (GenSmart Codon Optimisation). The optimisation was done for expression in human cells or human T cells. See, for example, W02020024917A1 which is incorporated herein by reference in its entirety. An exemplary expression vector sequence for codon-optimised, constant region-stabilised TCR_A0427 is shown in SEQ ID NO: 170. Lentivirus collected from the transfected HEK293 cells were then used to transduce each TCR into T cells. Transduction efficiency and expression of the introduced TCR were confirmed by flow cytometry.

[0726] Alternatively, the TCR was transfected into Jurkat cells using mRNA technology. mRNA was produced via IVT reaction (HiScribe® T7 mRNA Kit with CleanCap® Reagent AG, New England Biolabs) with template from restriction enzyme linearised plasmids containing T7 promoter followed by TCR beta and TCR alpha variable and constant regions. mRNA was transfected into Jurkat TCRap-knockout, CD8+ reporter T cells using a Neon™ Electroporation System. Expression of the TCR was validated and quantified by staining with anti-TCRap antibody followed by flow cytometry. Activation of successfully transfected Jurkat cells was tested in a dose response experiment, using a range of peptides presented on antigen-presenting cells (APCs), i.e., Raji cells stably transfected with A*01 :01 , A*03:01 , A*02:01 , A*24:02 or B*07:02.

[0727] Dose-response of transduced Jurkat reporter cells. Antigen presenting cells expressing a selected HLA allele were seeded into 96 well plates at 50,000 cells per well. In a separate plate, peptide dilutions were prepared to the desired range of concentrations for dose-response testing. 50,000 Jurkat reporter cells that were transduced or non-transduced with the TCR to be tested were added to the antigen presenting cells; transduction efficiency was quantified by flow cytometry, to verify that at least 10% of Jurkat reporter cells express the transduced TCR. Plates were incubated at 37°C in a cell culture incubator for 3h 50min with a final 10min of incubation at room temperature (4h incubation in total). To reveal the luciferase signal, Bio-Glo-NL™ reagent was added to all wells, reacted for 5 min and luminescence read on a SpectraMax i3x (Molecular Devices) plate reader.

[0728] Dose-response of transduced primary T cells. PBMCs from healthy donors (donor A and donor B) were used as a source of primary T cells for the preparation of effector cells. CD4+ and CD8+ T cells were positively enriched from PBMC using either A) CD4 and CD8 microbeads, and LS columns and magnets from Miltenyi or B) CliniMACS® CD4 and CD8 GMP MicroBeads, CliniMACS tubing set on the Miltenyi CliniMACS automated cell processing system. Cells were resuspended in AIM V medium with 10% heat-inactivated human AB serum and 10ng / ml IL-15. For T cell activation, cells were incubated with TransAct™ beads (Miltenyi). On the same or the next day, lentiviruses encoding for the TCR of interest were added to the activated T cells and incubated for three days. Cells that were not transduced with lentivirus and were used as control cells. Transduction efficiency was quantified by flow cytometry prior to performing cell-based functional assays. Effector cells (TCR-T or no-transduced T cells) were seeded in a U-bottom 96-well plate with Raji A02:01+ cells at the E / T ratio of 1 / 1 in the presence of WT-I37-45 peptide at a series of concentrations. Then the cell mixture was cultured for 18-26 hours and 150 pL supernatant was collected for interferon gamma (IFNy) release by ELISA analysis.

[0729] 1.2. Results

[0730] Jurkat reporter cells transduced with TCRs A0426, A0427, A0428, A0429, A0431 and A0432 demonstrated target peptide dose-dependent T cell activation upon incubation with Raji A*02:01 cells pulsed with WT-I37-45 (SEQ ID NO:1) as shown in Figure 1A and Figure 1B. In contrast, non-transduced Jurkat cells (NT) did not display the same dose-dependent T cell activation.

[0731] Jurkat reporter cells electroporated with mRNAs for TCRs A0426, A0427, A0428, A0431 and A0464 demonstrated target peptide dose-dependent T cell activation upon incubation with Raji A*02:01 cells pulsed with WT-I37-45 (SEQ ID NO:1) as shown in Figure 1C and Figure 1D.

[0732] Jurkat reporter cells expressing tested TCRs (e.g., A0426, A0427, A0428, A0429, A0431 , A0464) demonstrated increased target peptide T cell activation upon incubation with Raji A*02:01 cells pulsed with WT-137-45 (SEQ ID NO:1) as compared to reporter cells expressing a benchmark TCR A0418 (a known WT-137-45 targeting TCR disclosed in US11597755B2) (Figure 1A, Figure 1B, Figure 1C).

[0733] Primary T cells transduced with TCRs A0426, A0427, A0429, A0430, A0431 and A0432 demonstrated target peptide dose-dependent IFNy release upon incubation with Raji A*02:01 pulsed with a range of WT-137-45 (SEQ ID NO:1) (0.00005-50 pM, 10-fold dilution) as exemplified in Figure 1E. Values plotted have been adjusted against condition-matched controls using non-transduced primary T cells from the same donor (not shown). Primary T cells transduced with tested TCRs (e.g.. A0431 , A0427) demonstrated increased target peptide IFNy release upon incubation with Raji A*02:01 pulsed with WT- 137-45 as compared to primary T cells expressing benchmark TCR A0418. Example 2: Cytotoxicity activity of WT-137-45 specific T cell receptors (TCRs)

[0734] 2. 1. Materials and Methods

[0735] Quantification of cytotoxicity. PBMCs from healthy donors (donor A and donor B) were used as a source of primary T cells for the preparation of effector cells. CD4+ and CD8+ T cells were positively enriched from PBMC using either A) CD4 and CD8 microbeads, and LS columns and magnets from Miltenyi or B) CliniMACS® CD4 and CD8 GMP MicroBeads, CliniMACS tubing set on the Miltenyi CliniMACS automated cell processing system. Cells were resuspended in AIM V medium with 10% heat- inactivated human AB serum and 10ng / ml IL-15. For T cell activation, cells were incubated with TransActTM beads (Miltenyi). On the same or the next day, lentiviruses encoding for the TCR of interest were added to the activated T cells and incubated for three days. Cells that were not transduced with lentivirus and were used as control cells. Transduction efficiency was quantified by flow cytometry prior to performing cell based functional assays.

[0736] Flow Cytometry-based cytotoxicity assay. Raji cells expressing HLA-A*02:01 were first labeled with CellTrace™ Violet (CTV) before being pulsed with a range of WT-I 37-45 peptide (SEQ ID NO:1) (0.00005- 50 pM, 10-fold dilution). TCR-transduced primary T cells were then introduced to the CTV-labeled, peptide-pulsed target cells at an Effector:Target cell ratio of 1 :1 . After overnight (18-24h) incubation, flow cytometry was used to detect the percentage of live target cells based on CTV stain and live-dead cell counterstains, 7AAD and Annexin V. A cytotoxicity index was calculated relative to the average live cell count found in target-only control wells, using the following formula:

[0737] AveTargetOnly — Test well

[0738] Cytotoxicity (%) = %

[0739] AveTargetOnly

[0740] Cytotoxicity with xCelligence impedance readout. Cancer cell lines endogenously expressing the target antigen of interest and the HLA allele that tested TCRs are restricted for, were used as target cells. As effector cells, primary CD4+ and CD8+ T cells transduced with the TCR to be tested were used. Non- TCR transduced T cells were used as control. Target cell lines were seeded in xCelligence 96-well E plates at 10,000-25,000 cells per well, depending on the cell line used. Effector cells were added the next day at the Effector:Target (E:T) ratio indicated in the figure legend, preparing each condition in duplicate. Impedance readings were recorded for up to 90 hours. The percentage of cytolysis per condition and time point was calculated with XCelligence Immunotherapy software.

[0741] Quantification of cytokine secretion. IFNy secreted by T cells into the culture medium during the cytotoxicity assay was quantified by ELISA methodology. The OptEIA IFNy Kit from Becton Dickinson (BD) was used. Absorbance was measured with the SpectraMax i3x (Molecular Devices) plate reader and cytokine concentration calculated based on standard curve using SoftMax Pro 7.1.2 software.

[0742] 2.2. Results

[0743] Specific and direct cell killing by primary T cells transduced with TCRs A0427, A0430 and A0431 was measured by flow cytometry. TCR-Ts (TCR-transduced primary human T cells) demonstrated dosedependent cytotoxicity against Raji HLA-A*02:01 cells pulsed with a range of WT-I37-45 (SEQ ID NO:1) (0.00005-50 pM, 10-fold dilution) as exemplified in Figure 2A. These cytotoxicity activities were specific as non-transduced primary T cells (NT) from the same donor did not display a dose-dependent activity profile. Instead, NT controls showed a consistent background level of cytotoxicity that does not rise above that seen in WT-137-45 specific TCR-Ts at the lowest peptide concentration used.

[0744] The ability of TCR-transduced primary human T cells to kill tumor cells expressing a physiological quantity of WT-I37-45 (SEQ ID NO:1) in complex with HLA-A*02:01 molecules was assessed. Chronic Myelogenous Leukemia (CML) cell line K562, ovarian cancer cell line COV434 and glioblastoma cell line SF539, which are known to express WT-1 were employed. In some experiments, cell lines K562 and COV434 were also transduced with HLA-A*02:01 to enable the endogenous presentation of peptides on HLA*02:01 for the testing of HLA-A*02:01-restricted TCRs. IFNy secretion is known as a readout to quantify the cytotoxic activity of T cells (Murphy and Weaver, Janeway’s Immunobiology 9).

[0745] Figure 2B demonstrates the cell killing activity of TCR A0431 -transduced primary T cells against CML cell line K562 expressing HLA-A*02:01 at E:T ratios of 0.3:1 , 1 :1 and 3:1 . Non-transduced T cells from the same donor (NT) account for any non-target specific levels of IFNy release.

[0746] Figure 2C demonstrates the cell killing activity of TCR A0431 -transduced primary T cells against ovarian cancer cell line COV434 expressing HLA-A*02:01 at E:T ratios of 1 :1 and 3:1. Non-transduced T cells from the same donor (NT) account for any non-target specific levels of IFNy release.

[0747] Figure 2D demonstrates the cell killing activity of TCR A0431 -transduced primary T cells against glioblastoma cell line SF539 at E:T ratios of 0.3:1 , 1 :1 and 3:1 . Non-transduced T cells from the same donor (NT) account for any non-target specific levels of IFNy release.

[0748] These results demonstrate that endogenous levels of WT-I37-45 (SEQ ID NO:1) presentation on cell lines of different cancer origin were able to induce IFNy secretion from primary T cells transduced with A0431 . Therefore, A0431 -transduced T cells exhibit cell killing activities towards cancer cell lines displaying the WT-1 antigen of interest on the corresponding HLA molecule. In addition, the results showed A0431- transduced T cells exhibit improved cell killing activities compared to benchmark TCR A0418.

[0749] In addition, direct cell killing activities of TCR_A0427 or A0431 transduced primary T cells against ovarian cancer cell line COV434, transduced with HLA-A*02:01 , in Figure 2E, and glioblastoma cell line SF539 in Figure 2F, was measured using xCELLigence technology. Demonstrable cytotoxicity by both TCRs was measured over a 151 -hour period at 1 :1 and 3:1 E:T ratio.

[0750] Example 3: Alanine scan and safety asses...

Claims

1. Claims:1 . A T cell receptor (TCR) comprising a TCRa chain variable domain comprising a CDR3a having an amino acid sequence of SEQ ID NO:34, 10, 4, 16, 22, 28, 40 or 46, or a variant thereof in which 1 or 2 or 3 amino acids are substituted with another amino acid; in combination with a TCRp chain variable domain comprising a CDR3p having at least 95% sequence identity to an amino acid sequence of SEQ ID NO:37, 13, 7, 19, 25, 31 , 43 or 49, or a variant thereof in which 1 or 2 or 3 amino acids are substituted with another amino acid.

2. A TCR according to claim 1 , wherein the TCR comprises:(a)(i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NO:34, and / or(ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:37;(b)(i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NQ:10, and / or(ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:13;(c)(i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NO:4, and / or(ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NOT;(d)(i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NO:16, and / or(ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:19;(e)(i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NO:22, and / or(ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:25;(f)(i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NO:28, and / or(ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:31 ;(g)(i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NQ:40, and / or(ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQID NO:43; or(h)(i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NO:46, and / or(ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:49.

3. A TCR according to claim 1 or claim 2, wherein the TCR comprises:(a)(i) a TCRa chain variable domain incorporating the following CDRs:CDR1a having the amino acid sequence of SEQ ID NO:32 CDR2a having the amino acid sequence of SEQ ID NO:33CDR3a having the amino acid sequence of SEQ ID NO:34, and / or(ii) a TCRp chain variable domain incorporating the following CDRs:CDR1 p having the amino acid sequence of SEQ ID NO:35 CDR2p having the amino acid sequence of SEQ ID NO:36 CDR3p having the amino acid sequence of SEQ ID NO:37;(b)(i) a TCRa chain variable domain incorporating the following CDRs:CDR1a having the amino acid sequence of SEQ ID NO:8 CDR2a having the amino acid sequence of SEQ ID NO:9 CDR3a having the amino acid sequence of SEQ ID NQ:10, and / or(ii) a TCRp chain variable domain incorporating the following CDRs:CDR1 p having the amino acid sequence of SEQ ID NO:11 CDR2p having the amino acid sequence of SEQ ID NO:12 CDR3p having the amino acid sequence of SEQ ID NO:13;(c)(i) a TCRa chain variable domain incorporating the following CDRs:CDR1a having the amino acid sequence of SEQ ID NO:2 CDR2a having the amino acid sequence of SEQ ID NO:3 CDR3a having the amino acid sequence of SEQ ID NO:4, and / or(ii) a TCRp chain variable domain incorporating the following CDRs:CDR1 p having the amino acid sequence of SEQ ID NO:5 CDR2p having the amino acid sequence of SEQ ID NO:6 CDR3p having the amino acid sequence of SEQ ID NOT;(d)(i) a TCRa chain variable domain incorporating the following CDRs:CDR1a having the amino acid sequence of SEQ ID NO:14 CDR2a having the amino acid sequence of SEQ ID NO:15CDR3a having the amino acid sequence of SEQ ID NO:16, and / or(ii) a TCRp chain variable domain incorporating the following CDRs:CDR1 p having the amino acid sequence of SEQ ID NO:17 CDR2p having the amino acid sequence of SEQ ID NO:18CDR3p having the amino acid sequence of SEQ ID NO:19;(e)(i) a TCRa chain variable domain incorporating the following CDRs:CDR1a having the amino acid sequence of SEQ ID NQ:20CDR2a having the amino acid sequence of SEQ ID NO:21CDR3a having the amino acid sequence of SEQ ID NO:22, and / or(ii) a TCRp chain variable domain incorporating the following CDRs:CDR1 p having the amino acid sequence of SEQ ID NO:23 CDR2p having the amino acid sequence of SEQ ID NO:24 CDR3p having the amino acid sequence of SEQ ID NO:25;(f)(i) a TCRa chain variable domain incorporating the following CDRs:CDR1a having the amino acid sequence of SEQ ID NO:26CDR2a having the amino acid sequence of SEQ ID NO:27CDR3a having the amino acid sequence of SEQ ID NO:28, and / or(ii) a TCRp chain variable domain incorporating the following CDRs:CDR1 p having the amino acid sequence of SEQ ID NO:29 CDR2p having the amino acid sequence of SEQ ID NQ:30 CDR3p having the amino acid sequence of SEQ ID NO:31 ;(g)(i) a TCRa chain variable domain incorporating the following CDRs:CDR1a having the amino acid sequence of SEQ ID NO:38CDR2a having the amino acid sequence of SEQ ID NO:39CDR3a having the amino acid sequence of SEQ ID NQ:40, and / or(ii) a TCRp chain variable domain incorporating the following CDRs:CDR1 p having the amino acid sequence of SEQ ID NO:41CDR2p having the amino acid sequence of SEQ ID NO:42CDR3p having the amino acid sequence of SEQ ID NO:43; or(h)(i) a TCRa chain variable domain incorporating the following CDRs:CDR1a having the amino acid sequence of SEQ ID NO:44CDR2a having the amino acid sequence of SEQ ID NO:45CDR3a having the amino acid sequence of SEQ ID NO:46, and / or(ii) a TCRp chain variable domain incorporating the following CDRs:CDR1 p having the amino acid sequence of SEQ ID NO:47 CDR2p having the amino acid sequence of SEQ ID NO:48 CDR3p having the amino acid sequence of SEQ ID NO:49.1084. A TCR according to any one of claims 1 to 3, wherein the TCR comprises a TCRa chain variable domain comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:60, 52, 50, 54, 56, 58, 62 or 64.

5. A TCR according to any one of claims 1 to 4, wherein the TCR comprises a TCRp chain variable domain comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:61 , 53, 51 , 55, 57, 59, 63 or 65.

6. A TCR according to any one of claims 1 to 5, wherein the TCR is capable of binding to VX1DFX2X3PX4X5 (SEQ ID NO:181), VLDFX1PPX2X3 (SEQ ID NO:179), VLDFX1PX2GX3 (SEQ ID NO:182), and / or VLDFX1X2PX3X4 (SEQ ID NQ:180) when presented by an MHC class I molecule.

7. A TCR according to any one of claims 1 to 6, wherein the TCR is capable of binding to VX1DFX2X3PX4X5 (SEQ ID NQ:181)-HLA-A*02 complex, VLDFX1PPX2X3 (SEQ ID NQ:179)-HLA-A*02 complex, VLDFX1PX2GX3 (SEQ ID NQ:182)-HLA-A*02 complex, and / or VLDFX1X2PX3X4 (SEQ ID NQ:180)-HLA-A*02 complex.

8. A TCR according to any one of claims 1 to 7, wherein the TCR is capable of binding to VX1DFX2X3PX4X5 (SEQ ID NQ:181)-HLA-A*02:01 complex, VLDFX1PPX2X3 (SEQ ID NO:179)-HLA- A*02:01 complex, VLDFX1PX2GX3 (SEQ ID NQ:182)-HLA-A*02:01 complex, and / or VLDFX1X2PX3X4 (SEQ ID NQ:180)-HLA-A*02:01 complex.

9. A TCR according to any one of claims 1 to 8, wherein the TCR is capable of binding to VLDFAPPGA (SEQ ID NO:1) when presented by an MHC class I molecule.

10. A TCR according to any one of claims 1 to 9, wherein the TCR is capable of binding to a VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02 complex.

11. A TCR according to any one of claims 1 to 10, wherein the TCR is capable of binding to a VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02:01 complex, a VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02:07 complex, and / or a VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02:02 complex.

12. A T cell receptor (TCR) comprising a TCRa chain variable domain and a TCRp chain variable domain, wherein the TCR is capable of binding to VX1 DFX2X3PX4X5 (SEQ ID NO:181), VLDFX1PPX2X3 (SEQ ID NO:179), VLDFX1PX2GX3 (SEQ ID NO:182), and / or VLDFX1X2PX3X4 (SEQ ID NQ:180), when presented by an MHC class I molecule.

13. A TCR according to claim 12, wherein the TCR is capable of binding to VX1DFX2X3PX4X5 (SEQ ID NQ:181)-HLA-A*02 complex, VLDFX1PPX2X3 (SEQ ID NQ:179)-HLA-A*02 complex, VLDFX1PX2GX3 (SEQ ID NQ:182)-HLA-A*02 complex, and / or VLDFX1X2PX3X4 (SEQ ID NQ:180)-HLA-A*02 complex.10914. A TCR according to claim 12 or claim 13, wherein the TCR is capable of binding to VX1DFX2X3PX4X5 (SEQ ID N0:181)-HLA-A*02:01 complex, VLDFX1PPX2X3 (SEQ ID NO:179)-HLA- A*02:01 complex, VLDFX1PX2GX3 (SEQ ID NQ:182)-HLA-A*02:01 complex, and / or VLDFX1X2PX3X4 (SEQ ID NQ:180)-HLA-A*02:01 complex.

15. A TCR according to any one of claims 12 to 14, wherein the TCR is capable of binding to VLDFAPPGA (SEQ ID NO:1) when presented by an MHC class I molecule.

16. A TCR according to any one of claims 12 to 15, wherein the TCR is capable of binding to a VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02 complex.

17. A TCR according to any one of claims 12 to 16, wherein the TCR is capable of binding to a VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02:01 complex, a VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02:07 complex, and / or a VLDFAPPGA (SEQ ID NQ:1)-HLA-A*02:02 complex.

18. A TCR according to any one of claims 12 to 17, wherein the TCR comprises a TCRa chain variable domain comprising a CDR3a having an amino acid sequence of SEQ ID NO:34, 10, 4, 16, 22, 28, 40 or 46, or a variant thereof in which 1 or 2 or 3 amino acids are substituted with another amino acid; in combination with a TCRp chain variable domain comprising a CDR3p having an amino acid sequence of SEQ ID NO:37, 13, 7, 19, 25, 31 , 43 or 49, or a variant thereof in which 1 or 2 or 3 amino acids are substituted with another amino acid, respectively.

19. A TCR according to any one of claims 12 to 18, wherein the TCR comprises:(a)(i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NO:34, and / or(ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:37;(b)(i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NQ:10, and / or(ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:13;(c)(i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NO:4, and / or(ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NOT;(d)(i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NO:16, and / or(ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:19;(e)(i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NO:22, and / or(ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:25;(f)(i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NO:28, and / or(ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:31 ;(g)(i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NQ:40, and / or(ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:43; or(h)(i) a TCRa chain variable domain comprising a CDR3a having the amino acid sequence of SEQ ID NO:46, and / or(ii) a TCRp chain variable domain comprising a CDR3p having the amino acid sequence of SEQ ID NO:49.

20. A TCR according to any one of claims 12 to 19, wherein the TCR comprises:(a)(i) a TCRa chain variable domain incorporating the following CDRs:CDR1a having the amino acid sequence of SEQ ID NO:32CDR2a having the amino acid sequence of SEQ ID NO:33CDR3a having the amino acid sequence of SEQ ID NO:34, and / or(ii) a TCRp chain variable domain incorporating the following CDRs:CDR1 p having the amino acid sequence of SEQ ID NO:35CDR2p having the amino acid sequence of SEQ ID NO:36CDR3p having the amino acid sequence of SEQ ID NO:37;(b)(i) a TCRa chain variable domain incorporating the following CDRs:CDR1a having the amino acid sequence of SEQ ID NO:8CDR2a having the amino acid sequence of SEQ ID NO:9CDR3a having the amino acid sequence of SEQ ID NQ:10, and / or(ii) a TCRp chain variable domain incorporating the following CDRs:CDR1 p having the amino acid sequence of SEQ ID NO:11CDR2p having the amino acid sequence of SEQ ID NO:12CDR3p having the amino acid sequence of SEQ ID NO:13;(c)(i) a TCRa chain variable domain incorporating the following CDRs:CDR1a having the amino acid sequence of SEQ ID NO:2CDR2a having the amino acid sequence of SEQ ID NO:3CDR3a having the amino acid sequence of SEQ ID NO:4, and / or(ii) a TCRp chain variable domain incorporating the following CDRs:CDR1 p having the amino acid sequence of SEQ ID NO:5CDR2p having the amino acid sequence of SEQ ID NO:6 CDR3p having the amino acid sequence of SEQ ID NO:7;(d)(i) a TCRa chain variable domain incorporating the following CDRs:CDR1a having the amino acid sequence of SEQ ID NO:14 CDR2a having the amino acid sequence of SEQ ID NO:15 CDR3a having the amino acid sequence of SEQ ID NO:16, and / or(ii) a TCRp chain variable domain incorporating the following CDRs:CDR1 p having the amino acid sequence of SEQ ID NO:17 CDR2p having the amino acid sequence of SEQ ID NO:18 CDR3p having the amino acid sequence of SEQ ID NO:19;(e)(i) a TCRa chain variable domain incorporating the following CDRs:CDR1a having the amino acid sequence of SEQ ID NQ:20CDR2a having the amino acid sequence of SEQ ID NO:21CDR3a having the amino acid sequence of SEQ ID NO:22, and / or(ii) a TCRp chain variable domain incorporating the following CDRs:CDR1 p having the amino acid sequence of SEQ ID NO:23 CDR2p having the amino acid sequence of SEQ ID NO:24 CDR3p having the amino acid sequence of SEQ ID NO:25;(f)(i) a TCRa chain variable domain incorporating the following CDRs:CDR1a having the amino acid sequence of SEQ ID NO:26 CDR2a having the amino acid sequence of SEQ ID NO:27 CDR3a having the amino acid sequence of SEQ ID NO:28, and / or(ii) a TCRp chain variable domain incorporating the following CDRs:CDR1 p having the amino acid sequence of SEQ ID NO:29 CDR2p having the amino acid sequence of SEQ ID NQ:30 CDR3p having the amino acid sequence of SEQ ID NO:31 ;(g)(i) a TCRa chain variable domain incorporating the following CDRs:CDR1a having the amino acid sequence of SEQ ID NO:38 CDR2a having the amino acid sequence of SEQ ID NO:39 CDR3a having the amino acid sequence of SEQ ID NQ:40, and / or(ii) a TCRp chain variable domain incorporating the following CDRs:CDR1 p having the amino acid sequence of SEQ ID NO:41 CDR2p having the amino acid sequence of SEQ ID NO:42 CDR3p having the amino acid sequence of SEQ ID NO:43; or(h)(i) a TCRa chain variable domain incorporating the following CDRs:CDR1a having the amino acid sequence of SEQ ID NO:44 CDR2a having the amino acid sequence of SEQ ID NO:45 CDR3a having the amino acid sequence of SEQ ID NO:46, and / or(ii) a TCRp chain variable domain incorporating the following CDRs:CDR1 p having the amino acid sequence of SEQ ID NO:47 CDR2p having the amino acid sequence of SEQ ID NO:48 CDR3p having the amino acid sequence of SEQ ID NO:49.

21. A TCR according to any one of claims 12 to 20, wherein the TCR comprises a TCRa chain variable domain comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NQ:60, 52, 50, 54, 56, 58, 62 or 64.

22. A TCR according to any one of claims 12 to 21 , wherein the TCR comprises a TCRp chain variable domain comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:61 , 53, 51 , 55, 57, 59, 63 or 65.

23. A TCR according to any one of claims 1 to 22, wherein the TCR comprises a TCRa chain variable domain encoded by a nucleotide sequence having at least 80% sequence identity to SEQ ID NO:76, 92, 68, 84, 66, 70, 72, 74, 78, 80, 82, 86, 88, 90, 94 or 96.

24. A TCR according to any one of claims 1 to 23, wherein the TCR comprises a TCRp chain variable domain encoded by a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 77, 93, 69, 85, 67, 71 , 73, 75, 79, 81 , 83, 87, 89, 91 , 95 or 97.

25. A TCR according to any one of claims 1 to 24, wherein the TCR comprises a TCRa chain variable domain encoded by a nucleotide sequence having at least 80% sequence identity to SEQ ID NO:76, 92, 68, 84, 66, 70, 72, 74, 78, 80, 82, 86, 88, 90, 94 or 96; in combination with a TCRp chain variable domain encoded by a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 77, 93, 69, 85, 67, 71 , 73, 75, 79, 81 , 83, 87, 89, 91 , 95 or 97.

26. A TCR according to any one of claims 1 to 25, wherein the TCR comprises:(a)(i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:76 in combination with(ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:77;(b)113(i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:92 in combination with(ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:93;(c)(i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:68 in combination with(ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:69;(d)(i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:84 in combination with(ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:85;(e)(i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:66 in combination with(ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:67;(f)(i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NQ:70 in combination with(ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:71 ;(g)(i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:72 in combination with(ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:73;(h)(i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:74 in combination with(ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:75;(i)(i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:78 in combination with(ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:79;C)(i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NQ:80 in combination with(ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:81 ;(k)(i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:86 in combination with(ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:87;(l)(i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:88 in combination with(ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:89;(m)(i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:90 in combination with(ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:91 ;(n)(i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:94 in combination with(ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:95;(o)(i) a TCRa chain variable domain encoded by a nucleotide sequence of SEQ ID NO:96 in combination with(ii) a TCRp chain variable domain encoded by a nucleotide sequence of SEQ ID NO:97.

27. A TCR according to any one of claims 1 to 26, wherein the TCR comprises a TCRa chain constant domain comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:162 or 165.

28. A TCR according to any one of claims 1 to 27, wherein the TCR comprises a TCRp chain constant domain comprising an amino acid sequence having at least 80% sequence identity to an amino acid sequence selected from SEQ ID NO:163, 164 or 166.

29. A multispecific antigen-binding molecule comprising a TCR or an antigen-binding fragment of a TCR according to any one of claims 1 to 28.

30. A multispecific antigen-binding molecule according to claim 23, further comprising an antigenbinding moiety capable of binding to a molecule expressed by an immune cell, optionally wherein the molecule expressed by an immune cell is CD3.31 . A nucleic acid comprising a nucleotide sequence encoding all or part of a TCR according to any one of claims 1 to 28.

32. An isolated nucleic acid comprising a nucleotide sequence having at least 80% sequence identity to:(a) SEQ ID NO:76, 77, 92 and / or 93;(b) SEQ ID NO:68, 69, 84 and / or 85;(c) SEQ ID NO:66, 67, 82 and / or 83;(d) SEQ ID NQ:70, 71 , 86 and / or 87;(e) SEQ ID NO:72, 73, 88 and / or 89;(f) SEQ ID NO:74, 75, 90 and / or 91 ;(g) SEQ ID NO:78, 79, 94 and / or 95; or(h) SEQ ID NQ:80, 81 , 96 and / or 97.11533. An isolated nucleic acid comprising a nucleotide sequence having at least 80% sequence identity to:(a) SEQ ID NO:92 and / or 93;(b) SEQ ID NO:84 and / or 85;(c) SEQ ID NO:82 and / or 83;(d) SEQ ID NO:86 and / or 87;(e) SEQ ID NO:88 and / or 89;(f) SEQ ID NQ:90 and / or 91 ;(g) SEQ ID NO:94 and / or 95; or(h) SEQ ID NO:96 and / or 97.

34. A nucleic acid comprising a nucleotide sequence encoding all or part of a multispecific antigenbinding molecule according to claim 29 or 30.

35. A vector comprising a nucleic acid or nucleotide sequence according to any one of claims 31 to 34.

36. A vector according to claim 35, further comprising a nucleotide sequence of SEQ ID NO:171 .

37. A host cell comprising and / or expressing: a TCR according to any one of claims 1 to 28, a multispecific antigen-binding molecule according to claim 29 or 30, a nucleic acid according to any one of claims 31 to 34, and / or a vector according to claim 35 or 36.

38. A host cell according to claim 37, wherein the host cell is an immune cell.

39. A host cell according to claim 37 or claim 38, wherein the host cell is a T lymphocyte.

40. A composition comprising a TCR according to any one of claims 1 to 28, a multispecific antigenbinding molecule according to claim 29 or 30, a nucleic acid according to any one of claims 31 to 34, a vector according to claim 35 or 36, or a host cell according to any one of claims 37 to 39.41 . A composition according to claim 40, wherein the composition further comprises a pharmaceutical carrier or excipient.

42. A TCR according to any one of claims 1 to 28, a multispecific antigen-binding molecule according to claim 29 or 30, a nucleic acid according to any one of claims 31 to 34, a vector according to claim 35 or 36, a host cell according to any one of claims 37 to 39, or a composition according to claim 40 or 41 , for use as a medicament.

43. A method of treating a disease or condition associated with expression of WT-1 , comprising administering a TCR according to any one of claims 1 to 28, a multispecific antigen-binding moleculeaccording to claim 29 or 30, a nucleic acid according to any one of claims 31 to 34, a vector according to claim 35 or 36, a host cell according to any one of claims 37 to 39, or a composition according to claim 40 or 41 to a subject.

44. A TCR according to any one of claims 1 to 28, a multispecific antigen-binding molecule according to claim 29 or 30, a nucleic acid according to any one of claims 31 to 34, a vector according to claim 35 or 36, a host cell according to any one of claims 37 to 39, or a composition according to claim 40 or 41 , for use in a method of treating a disease or condition associated with expression of WT-1 .

45. Use of a TCR according to any one of claims 1 to 28, a multispecific antigen-binding molecule according to claim 29 or 30, a nucleic acid according to any one of claims 31 to 34, a vector according to claim 35 or 36, a host cell according to any one of claims 37 to 39, or a composition according to claim 40 or 41 , in the manufacture of a medicament for the treatment of a disease or condition associated with expression of WT-1 .

46. The method according to claim 43, the TCR, multispecific antigen-binding molecule, nucleic acid, vector, cell or composition for use according to claim 44, or the use according to claim 45, wherein the disease or condition is cancer or a neoplasm.

47. The method, agent for use, or the use according to any one of claims 43 to 46, wherein the disease or condition is selected from: a WT-1 -associated cancer, a cancer comprising cells comprising / presenting a peptide of a WT-1 antigen, a cancer comprising cells comprising / presenting a peptide of SEQ ID NO:1 , 179, 180, 181 or 182, a cancer comprising cells comprising / presenting a peptide of WT-1 and a major histocompatibility complex (MHC) molecule comprising an MHC class I molecule encoded by a HLA-A*02:01 allele, and a cancer comprising cells comprising / presenting a peptide of SEQ ID NO:1 , 179, 180, 181 or 182, and a major histocompatibility complex (MHC) molecule comprising an MHC class I molecule encoded by a HLA-A*02:01 allele, a cancer comprising cells comprising / presenting a peptide of WT-1 and a major histocompatibility complex (MHC) molecule comprising an MHC class I molecule encoded by a HLA-A*02:07 allele, a cancer comprising cells comprising / presenting a peptide of SEQ ID NO:1 , 179, 180, 181 or 182, and a major histocompatibility complex (MHC) molecule comprising an MHC class I molecule encoded by a HLA-A*02:07 allele, a cancer comprising cells comprising / presenting a peptide of WT-1 and a major histocompatibility complex (MHC) molecule comprising an MHC class I molecule encoded by a HLA-A*02:02 allele, a cancer comprising cells comprising / presenting a peptide of SEQ ID NO:1 , 179, 180, 181 or 182, and a major histocompatibility complex (MHC) molecule comprising an MHC class I molecule encoded by a HLA-A*02:02 allele.

48. The method, agent for use, or the use according to any one of claims 43 to 47, wherein the disease or condition is selected from: a solid cancer, a hematological cancer, gastric cancer (e.g., gastric carcinoma, gastric adenocarcinoma, gastrointestinal cancer, gastrointestinal adenocarcinoma), liver cancer (hepatocellular carcinoma, cholangiocarcinoma), head and neck cancer (e.g., head and neck squamous cell carcinoma), oral cavity cancer (e.g., oropharyngeal cancer (e.g., oropharyngeal carcinoma), oral cancer, oral squamous cell carcinoma (OSCC), laryngeal cancer, nasopharyngealcarcinoma, oesophageal cancer), colorectal cancer (e.g., colorectal carcinoma), colon cancer, colon carcinoma, cervical carcinoma, prostate cancer, lung cancer (e.g., NSCLC, small cell lung cancer, lung adenocarcinoma, squamous lung cell carcinoma), bladder cancer, urothelial carcinoma, urogenital cancer, skin cancer (e.g., melanoma, advanced melanoma), renal cell cancer (e.g., renal cell carcinoma), ovarian cancer (e.g., ovarian carcinoma), mesothelioma, breast cancer (e.g., triple-negative, Luminal A, Luminal B, and / or HER2+ breast cancer), brain cancer (e.g., glioblastoma), prostate cancer, pancreatic cancer, endometrial cancer, a myeloid hematologic malignancy, a lymphoblastic hematologic malignancy, myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute lymphoblastic leukemia (ALL), lymphoma, non-Hodgkin’s lymphoma (NHL), thymoma and multiple myeloma (MM).

49. The method, agent for use, or the use according to any one of claims 43 to 48, wherein the treatment further comprises administering one or more immune modulating agents to the subject, wherein the one or more immune modulating agents are selected from the group consisting of: immune celldepleting agents, cytokines, TLR agonists, RIG-1 like receptor (RLR) agonists, immune checkpoint inhibitors, chemotherapeutic agents, antibodies, radiotherapy, and a combination thereof.

50. The method, agent for use, or the use according to any one of claims 43 to 49, wherein the treatment comprises:(a) modifying a cell to express or comprise the TCR or a nucleic acid or vector encoding the TCR, and / or(b) administering a / the cell modified to express or comprise a TCR or nucleic acid or vector encoding the TCR to a subject.51 . The method, agent for use, or the use according to any one of claims 43 to 50, wherein the treatment comprises administering at least one TCR, multispecific antigen-binding molecule, nucleic acid, vector, cell and / or composition according to any preceding claim to a subject, in combination with at least one non-identical TCR, multispecific antigen-binding molecule, nucleic acid, vector, cell and / or composition.

52. The method, agent for use, or the use according to any one of claims 43 to 51 , wherein the subject comprises a cell that expresses WT-1 , and / or that expresses a WT-1 peptide of SEQ ID NO:1 , 179, 180, 181 or 182.

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