MODIFIED T-CELLS FOR USE IN THE TREATMENT OF GASTROESOPHAGEAL CANCER

Abstract

The disclosure relates to a method of treating gastroesophageal cancer, and to a population of modified T cells expressing a heterologous TCR for use in such method.

Claims

1. A method of treating gastroesophageal cancer in an individual, comprising administering to the individual a population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous T-cell receptor (TCR) capable of binding to a peptide antigen of MAGE-A4.

2. The method of claim 1, wherein the method further comprises administering an additional anti-cancer therapy to the individual, optionally wherein the population of modified T cells and the additional anti-cancer therapy are administered in the same line of therapy.

3. The method of claim 2, wherein: (a) administration of the additional anti-cancer therapy begins before administration of the population of modified T cells, and optionally continues after administration of the population of modified T cells; or (b) administration of the additional anti-cancer therapy begins after administration of the population of modified T cells.

4. The method of claim 2 wherein the anti-cancer therapy comprises a chemotherapy and/or a targeted therapy.

5. The method of claim 2, wherein the anti-cancer therapy comprises (i) paclitaxel and/or ramucirumab, or (ii) oxaliplatin.

6. The method of claim 1, wherein the method further comprises administering a checkpoint inhibitor to the individual, optionally wherein: (a) the population of modified T cells and the checkpoint inhibitor are administered in the same line of therapy; and/or (b) the checkpoint inhibitor comprises a Programmed Death-1 (PD-1) axis binding antagonist, optionally nivolumab or pembrolizumab.

7. The method of claim 6, wherein: (a) administration of the checkpoint inhibitor begins at the same time administration of the population of modified T cells and continues after administration of the population of modified T cells; and/or (b) the method comprises administering an additional anti-cancer therapy to the individual and administration of the checkpoint inhibitor begins after administration of the additional anti-cancer therapy and the population of modified T cells.

8. The method of claim 1, wherein the gastroesophageal cancer has relapsed following curative intent treatment for locally advanced cancer, or is the first diagnosis of unresectable locally advanced cancer or metastatic cancer.

9. The method of claim 8, wherein the method comprises: (a) administering oxaliplatin to the individual and administration of oxaliplatin begins before administration of the population of modified T cells and optionally continues after administration of the population of modified T cells; and (b) administering a checkpoint inhibitor and/or an additional anti-cancer therapy to the individual and administration of the checkpoint inhibitor and/or additional anti-cancer therapy begins after administration of the population of modified T cells, optionally wherein the checkpoint inhibitor is a PD-1 axis binding antagonist and/or the additional anti-cancer therapy is a chemotherapy such as fluorouracil.

10. The method of claim 1, wherein the gastroesophageal cancer is relapsed gastroesophageal cancer.

11. The method of claim 10, wherein the gastroesophageal cancer has relapsed following a first-line treatment for gastroesophageal cancer, optionally wherein: (a) the first-line treatment comprises surgical resection and/or radiation therapy; and/or (b) the first-line treatment comprises systemic therapy.

12. The method of claim 10, wherein: (a) the method comprises administering an additional anti-cancer therapy to the individual and administration of the additional anti-cancer therapy begins before administration of the population of modified T cells and continues after administration of the population of modified T cells, optionally wherein the method further comprises administering a checkpoint inhibitor to the individual and administration of the checkpoint inhibitor begins after administration of the additional anti-cancer therapy and the population of modified T cells; or (b) the method comprises administering an additional anti-cancer therapy to the individual and administration of the additional anti-cancer therapy begins after administration of the population of modified T cells, optionally wherein the method further comprises administering a checkpoint inhibitor to the individual and administration of the checkpoint inhibitor begins after administration of the additional anti-cancer therapy and the population of modified T cells; optionally wherein (i) the anti-cancer therapy comprises paclitaxel and ramucirumab, and/or (ii) the checkpoint inhibitor comprises nivolumab or pembrolizumab.

13. The method of claim 11, wherein the gastroesophageal cancer has further relapsed following a second-line treatment for gastroesophageal cancer, optionally wherein: (a) the second-line treatment comprises surgical resection and/or radiation therapy; and/or (b) the second-line treatment comprises systemic therapy.

14. The method of claim 13, wherein the method comprises administering a checkpoint inhibitor to the individual and (a) administration of the checkpoint inhibitor begins at the same time administration of the population of modified T cells and continues after administration of the population of modified T cells; or (b) administration of the checkpoint inhibitor begins after administration of the population of modified T cells; optionally wherein the checkpoint inhibitor comprises nivolumab or pembrolizumab.

15. The method of claim 1, wherein: (a) the heterologous TCR binds to GVYDGREHTV (SEQ ID NO: 1) in complex with an HLA molecule; (b) the heterologous TCR comprises an alpha chain amino acid sequence having at least 80% sequence identity to SEQ ID NO: 2 and a beta chain amino acid sequence having at least 80% sequence identity to SEQ ID NO: 3; and/or (c) the CD8 co-receptor is CD8.

16. The method of claim 1, wherein the modified T cells are autologous with respect to the individual, optionally wherein the method comprises producing the population by: (a) obtaining peripheral blood mononuclear cells (PBMCs) from the individual; (b) selecting T cells from the PBMCs; and (c) modifying the selected T cells to express the heterologous CD8 co-receptor and the heterologous TCR; further optionally wherein the gastroesophageal cancer is relapsed gastroesophageal cancer and wherein one or more of steps (a) to (c) are performed prior to relapse.

17. A population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous T-cell receptor capable of binding to a peptide antigen of MAGE-A4, for use in the method of claim 1.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0014] FIG. 1: Exemplary treatment regimens for gastroesophageal cancer that has relapsed following first-line treatment.

[0015] FIG. 2: Exemplary treatment regimen for gastroesophageal cancer that has relapsed following second-line treatment.

[0016] FIG. 3: Alternative representation of exemplary treatment regimens for (1) gastroesophageal cancer that has relapsed following first-line treatment and (2) gastroesophageal cancer that has relapsed following second-line treatment.

[0017] FIG. 4: Exemplary treatment regimens for gastroesophageal cancer that (A) has relapsed following curative intent treatment for locally advanced cancer, or (B) is the first diagnosis of unresectable locally advanced cancer or metastatic cancer.

[0018] FIG. 5: Alternative representation of exemplary treatment regimens for gastroesophageal cancer that (A) has relapsed following curative intent treatment for locally advanced cancer, or (B) is the first diagnosis of unresectable locally advanced cancer or metastatic cancer.

[0019] FIG. 6: Efficacy of ADP-A2M4CD8 in patients with advanced esophageal, esophagogastric junction, or gastric cancer. FIG. 6A: Change in baseline sum of longest diameters of target lesion (SLD) in individual patients. FIG. 6B: Change in baseline target SLD in weeks from infusion of T-cells. One non-evaluable patient not shown. Data show change from baseline in SLD through progression or prior to surgical resection. Investigator-assessed best overall response indicated per RECIST v1.1. Combo refers to patients in a nivolumab combination group. EGJ: esophagogastric junction. ESO: esophageal. G: gastric. PD: progressive disease. PR: partial response. RECIST: Response Evaluation Criteria in Solid Tumors. SD: stable disease.

DETAILED DESCRIPTION

[0020] It is to be understood that different applications of the disclosed methods and products may be tailored to the specific needs in the art. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments of the disclosure only, and is not intended to be limiting.

[0021] All publications, patents and patent applications cited herein, whether supra or infra, are hereby incorporated by reference in their entirety.

General Definitions

[0022] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by a person skilled in the art to which this disclosure belongs.

[0023] As used in this specification and the appended claims, the singular forms a, an, and the include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to a TCR includes TCRs, reference to an antibody includes two or more such antibodies, and the like.

[0024] In general, the term comprising is intended to mean including but not limited to. For example, the phrase a method comprising administering a population of modified T cells should be interpreted to mean that the method contains a step administering such a population, but that the method may contain additional such as, for example, administering a further therapeutic agent.

[0025] In some aspects of the disclosure, the word comprising is replaced with the phrase consisting of. The term consisting of is intended to be limiting. For example, the phrase a method consisting of administering a population of modified T cells should be interpreted to mean that the method contains a step administering such a population, and no additional steps.

[0026] The terms protein and polypeptide are used interchangeably herein, and are intended to refer to a polymeric chain of amino acids of any length.

[0027] Typically, the term about is used to refer to a value within +/10% (such as within +/5% or within +/2%) of the value that follows.

[0028] As used herein, the terms recurrent cancer and relapsed cancer are interchangeable.

[0029] For the purpose of this disclosure, in order to determine the percent identity of two sequences (such as two polynucleotide or two polypeptide sequences), the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in a first sequence for optimal alignment with a second sequence). The nucleotide residues at nucleotide positions are then compared. When a position in the first sequence is occupied by the same nucleotide residue as the corresponding position in the second sequence, then the nucleotides are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., % identity=number of identical positions/total number of positions in the reference sequence100).

[0030] Typically, the sequence comparison is carried out over the length of the reference sequence. For example, if the user wished to determine whether a given (test) sequence has a certain percentage identity to SEQ ID NO: X, SEQ ID NO: X would be the reference sequence. For example, to assess whether a sequence is at least 80% identical to SEQ ID NO: X (an example of a reference sequence), the skilled person would carry out an alignment over the length of SEQ ID NO: X, and identify how many positions in the test sequence were identical to those of SEQ ID NO: X. If at least 80% of the positions are identical, the test sequence is at least 80% identical to SEQ ID NO: X. If the sequence is shorter than SEQ ID NO: X, the gaps or missing positions should be considered to be non-identical positions.

[0031] The skilled person is aware of different computer programs that are available to determine the homology or identity between two sequences. For instance, a comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm.

Method of Treating Cancer

[0032] The disclosure provides a method of treating gastroesophageal cancer in an individual, comprising administering to the individual a population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous T-cell receptor (TCR) capable of binding to a peptide antigen of MAGE-A4. As set out above, the present inventors have identified that such a method is advantageous. Modified T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor capable of binding to a peptide antigen of MAGE-A4 may, for example, be comprised in a second- or third-line treatment for gastroesophageal cancer.

[0033] In the context of the disclosure, treating gastroesophageal cancer may also encompass preventing and/or delaying the progression of gastroesophageal cancer.

Gastroesophageal Cancer in an Individual

[0034] The method of the disclosure is for treating gastroesophageal cancer in an individual. The individual is preferably human. The individual may, for example, be a non-human mammal, such as a mouse, rat, rabbit, cat, dog, pig, cow or horse.

[0035] The gastroesophageal cancer may be a cancer that expresses MAGE-A4. MAGE-A4 expression has been reported in gastroesophageal cancer. For example, approximately 20% of solid tumours in gastroesophageal cancer express MAGE-A4, of which 40-45% of individuals also express HLA-A*02.

[0036] At least 1% of the gastroesophageal cancer cells from the individual may express MAGE-A4, such as at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70% or at least 80%. The percentage of cells that express MAGE-A4 may be determined by any means known to the skilled person, such as immunohistochemistry (IHC), flow-cytometry or enzyme-linked immunosorbent assay (ELISA).

[0037] The expression of MAGE-A4 in the gastroesophageal cancer may have an intensity of greater than or equal to () 1+, such as 2+ or 3+. The intensity score may be assessed by IHC staining of the tumour, with the scoring as follows: negative=no staining or staining in less than or equal to 10% of the cells stained; 1+=incomplete staining in 10% of cells stained; 2+=weak to moderate staining in 10% of cells stained; strong and complete staining in 10% of cells stained.

[0038] The gastroesophageal cancer may be a gastroesophageal tumour. The gastroesophageal cancer may, for example, be a solid tumour. The gastroesophageal cancer may, for example, be a squamous cell carcinoma, an adenocarcinoma, a gastrointestinal stromal tumour (GIST), a non-Hodgkin lymphoma, a neuroendocrine tumour (NET), a melanoma, a leiomyosarcoma, a carcinoid tumour or a lymphoma. The gastroesophageal cancer may, for example, be a squamous cell carcinoma or an adenocarcinoma.

[0039] The gastroesophageal cancer may, for example, be an esophageal cancer. The esophageal cancer may, for instance, be an esophageal squamous cell carcinoma (ESCC). The esophageal cancer may, for example, be esophageal adenocarcinoma (EAC). The gastroesophageal cancer may, for example, be an esophagogastric junction cancer (EGJ; also known as gastroesophageal junction cancer or GOJ). The gastroesophageal cancer may, for example, be a cervical esophagus cancer. The gastroesophageal cancer may, for example, be a gastric cancer. The gastric cancer may, for example, be gastric adenocarcinoma. The gastroesophageal cancer may be associated with gastroesophageal reflux disease (GERD) or Barrett's esophagus.

[0040] The gastroesophageal cancer may, for example, be primary or secondary gastroesophageal cancer. The gastroesophageal cancer may, for example, be recurrent or relapsed, unresectable, locally advanced, and/or metastatic gastroesophageal cancer. For example, the gastroesophageal cancer may not be suitable for treatment by surgical resection or radiotherapy.

[0041] The gastroesophageal cancer may be relapsed gastroesophageal cancer. The relapsed gastroesophageal cancer may, for instance, be a locally advanced relapse or a metastatic relapse. The gastroesophageal cancer may, for instance, have relapsed following curative intent treatment. Accordingly, the individual may be a cancer patient that has received curative intent treatment of gastroesophageal cancer. Such cancer patients form a subpopulation of gastroesophageal patients that is well-recognised in the art.

[0042] In the context of the disclosure, a line of therapy may refer to a treatment regimen for a cancer for which curative intent treatment has failed, or for which curative intent treatment may be inappropriate. Curative intent treatment may refer to any therapy that has the potential to cure the cancer in the individual. Curative intent treatment may, for example, refer to a therapy that is administered to the individual to try to cure the cancer. Curative intent treatment may precede a line of therapy for a cancer that has is recurrent or has spread.

[0043] Failure of a curative intent treatment may, for example, refer to failure of the curative intent treatment to eliminate the cancer or to induce remission. For instance, failure of the curative intent treatment may result in relapse, or in spread of the cancer such as local invasion or metastasis. A curative intent treatment may be inappropriate when the cancer is advanced, for instance when the cancer is a locally advanced cancer or a metastatic cancer. A line of therapy may therefore refer to a treatment regimen for a locally advanced, metastatic, or relapsed cancer. A line of therapy may refer to a treatment regimen for a cancer that is recurrent or have spread.

[0044] A line of therapy may, for example, be a first-line therapy for the cancer. In other words, the treatment regimen may be the first treatment regimen employed against the cancer after failure or curative intent treatment. For instance, the treatment regimen may be the first treatment regimen employed after recurrence or spread of the cancer. For example, the treatment regimen may be the first treatment regimen employed against the cancer after local advance/invasion, metastasis, or relapse. A line of therapy may, for example, be a second-line therapy for the cancer. In other words, the treatment regimen may be the second treatment regimen employed against the cancer, for instance after failure of the first treatment regimen. Failure of the first treatment regimen may, for example, result in recurrence or further spread of the cancer. Failure of the first treatment regimen may, for example, result in local advance/invasion, metastasis, or relapse. A line of therapy may, for example, be a third-line therapy for the cancer. In other words, the treatment regimen may be the third treatment regimen employed against the cancer, for instance after failure of the second treatment regimen. Failure of the second treatment regimen may, for example, result in recurrence or further spread of the cancer. Failure of the second treatment regimen may, for example, result in local advance/invasion, metastasis, or relapse.

[0045] The gastroesophageal cancer may, for example, have relapsed following a first-line treatment for gastroesophageal cancer. Relapse may, for example, refer to recurrence, local invasion, and/or metastasis. First-line treatments are described in detail below. However, by way of example, the first-line treatment may be a treatment previously described as an approved or standard-of-care first-line treatment for one or more gastroesophageal cancers. In this context, approval may relate to approval by the FDA, EMA or MHRA for example. Standard-of-care first-line treatments for gastroesophageal cancers are well-known in the art and described, for instance, in publicly-available clinical guidelines such as those provided by the National Comprehensive Cancer Network.

[0046] Standard-of-care first-line treatments for operable gastroesophageal cancer (or gastroesophageal cancers for which local therapy is indicated) may, for example, comprise (i) surgical resection, (ii) radiation therapy, and/or (iii) systemic therapy. For instance, standard-of-care first-line treatments for operable gastroesophageal cancer may comprise (i); (ii); (iii); (i) and (ii); (i) and (iii); (ii) and (iii); or (i), (ii) and (iii). Operable gastroesophageal cancers may, for example, include pTis tumours (primary tumours confined to epithelium by basement membrane) and pT1 tumours (primary tumours having invaded the lamina propria, muscularis mucosae, or submucosa). Surgical therapy for such cancers may, for example, comprise endoscopic resection, endoscopic resection followed by ablation, or esophagectomy.

[0047] Standard-of-care first-line treatments for inoperable gastroesophageal cancers (or gastroesophageal cancers where surgical approaches are less preferred, such as esophagogastric junction adenocarcinomas and cervical esophagus cancer) may, for example, comprise definitive chemoradiotherapy. Alternatively, other anti-cancer therapies (such as systemic therapies) may be used.

[0048] The gastroesophageal cancer to be treated by the method of the disclosure may have relapsed following a second-line or subsequent-line treatment for gastroesophageal cancer. In other words, the gastroesophageal cancer may have relapsed for a first time following a first-line treatment, and subsequently relapsed for a second time following a second-line treatment. The gastroesophageal cancer may further have lapsed to a subsequent-line treatment. Relapse may, for example, refer to recurrence, local invasion, and/or metastasis. First-line and second-line treatments are described in detail below. However, by way of example, the first-line treatment may be a treatment previously described as an approved or standard-of-care first-line treatment for one or more gastroesophageal cancers, as set out above. Also by way of example, the second-line treatment may be a treatment previously described as an approved or standard-of-care second-line treatment for one or more gastroesophageal cancers. In this context, approval may relate to approval by the FDA, EMA or MHRA for example. Standard-of-care second-line treatments for gastroesophageal cancers are well-known in the art and described, for instance, in publicly-available clinical guidelines such as those provided by the National Comprehensive Cancer Network.

[0049] Second-line standard-of-care treatments for gastroesophageal cancer may, for instance, comprise chemoradiotherapy, surgery, chemotherapy or other anti-cancer systemic therapies if the first-line standard-of-care treatment comprised a esophagectomy without chemoradiotherapy. Second-line standard-of-care treatments for gastroesophageal cancer may, for example, comprise surgery (where the cancer is operable) or other anti-cancer systemic therapies if the first-line standard-of-care treatment comprised chemoradiotherapy without surgery. Second-line standard-of-care treatments for gastroesophageal cancer may, for example, comprise an anti-cancer systemic therapy if metastatic gastroesophageal cancer followed the first-line standard-of-care treatment.

[0050] The gastroesophageal cancer to be treated by the method of the disclosure may, for example, be gastroesophageal cancer that (A) has relapsed following curative intent treatment for locally advanced cancer, or (B) is the first diagnosis of unresectable locally advanced cancer or metastatic cancer. Curative intent treatment are described in detail below. However, by way of example, the curative intent treatment may be a treatment previously described as an approved or standard-of-care curative intent treatment for one or more gastroesophageal cancers. In this context, approval may relate to approval by the FDA, EMA or MHRA for example. Standard-of-care curative intent treatments for gastroesophageal cancers are well-known in the art and described, for instance, in publicly-available clinical guidelines such as those provided by the National Comprehensive Cancer Network.

[0051] Accordingly, the individual may be a gastroesophageal cancer patient that has received treatment for gastroesophageal cancer. The individual may be a gastroesophageal cancer patient that has received curative intent treatment for gastroesophageal cancer. The individual may be a gastroesophageal cancer patient that has received a first-line treatment for gastroesophageal cancer. The individual may be a gastroesophageal cancer patient that has received a first-line treatment and a second-line treatment for gastroesophageal cancer. Such cancer patients form subpopulations of gastroesophageal cancer patients that are well-recognised in the art. Curative intent, first-line and second-line treatments are described in more detail below.

[0052] Gastroesophageal cancer may be prone to relapse. For instance, gastroesophageal cancer may be prone to recurrence, local invasion, and/or metastasis. The method of the disclosure may aim to treat the relapse, recurrence, local invasion, and/or metastasis.

Curative Intent Treatment

[0053] Curative intent treatment may refer to any therapy that has the potential to cure the cancer in the individual. Curative intent treatment may, for example, refer to a therapy that is administered to the individual to try to cure the cancer. Curative intent treatment may precede a line of therapy for a cancer that has is recurrent or has spread.

[0054] As explained above, known treatments for gastroesophageal cancer include surgical resection, radiation therapy and systemic therapy. A curative intent treatment (such as the first curative intent treatment or the second curative intent treatment) may comprise (i) surgical resection, (ii) radiation therapy, and/or (iii) systemic therapy. For example, a curative intent treatment may comprise (i); (ii); (iii); (i) and (ii); (i) and (iii); (ii) and (iii); or (i), (ii) and (iii). In particular aspects of the disclosure, a curative intent treatment may comprise (i); (ii); or (i) and (ii).

[0055] The systemic therapy may comprise or consist of (a) a chemotherapy. The systemic therapy may comprise or consist of (b) an immunotherapy. The systemic therapy may comprise or consist of (c) a targeted therapy. For example, the systemic therapy may comprise or consist of (a); (b); (c); (a) and (b); (a) and (c); (b) and (c); or (a), (b) and (c).

[0056] Chemotherapies, such as those for gastroesophageal cancer, are well-known in the art. Such chemotherapies may, for example, comprise a platinum-based anti-neoplastic drug, such as cisplatin, oxaliplatin or carboplatin. Such chemotherapies may, for example, comprise an anti-metabolite, such as fluorouracil (5-FU), trifluridine or capecitabine. Such chemotherapies may, for example, comprise a taxane drug, such as docetaxel or paclitaxel. Leucovorin may be administered with a chemotherapy to reduce the toxic effects of the chemotherapy.

[0057] Immunotherapies, such as those for gastroesophageal cancer, are well-known in the art. Such immunotherapies may, for example, include therapeutic immune cells, immunomodulators, checkpoint inhibitors, and vaccines. Therapeutic immune cells may include T cells, for instance engineered T cells such as CAR T cells or T cells expressing an engineered TCR. Immunomodulators may include, for example, interleukins, cytokines, chemokines, and immunomodulatory imide drugs. Checkpoint inhibitors may, for instance, include CTLA-4 inhibitors or PD-1 axis binding antagonists. CTLA-4 inhibitors may include ipilimumab. PD-1 axis binding antagonists may, for instance, include pembrolizumab, dostarlimab-gxly and nivolumab. Checkpoint inhibitors and PD-1 axis binding antagonists are described in detail below.

[0058] Targeted therapies, such as those for gastroesophageal cancer, are well-known in the art. The term targeted therapy is a term of art that refers to treatments that target specific genes and proteins that help cancer cells survive and grow. Targeted therapies may, for example, include a tropomysin kinase receptor antagonist (such as larotrectinib or entrectinib), a HER2 antagonist (such as trastuzumab), a topoisomerase I inhibitor (such as irinotecan or deruxtecan-nxki), a VEGF inhibitor (such as ramucirumab), or a thymidine phosphorylase inhibitor (such as tiparicil).

[0059] The systemic therapy may, for example, comprise a drug (or a combination of drugs) previously described for therapy of gastroesophageal cancer. The systemic therapy may comprise a drug, or a combination of drugs, previously described as an approved of standard-of-care curative treatment for gastroesophageal cancer.

[0060] For example, the systemic therapy may comprise a drug, or a combination of drugs, previously described for preoperative chemoradiotherapy or preoperative chemotherapy in standard-of-care curative intent treatment for operable gastroesophageal cancer (or gastroesophageal cancer for which local therapy is indicated). The systemic therapy may, for example, comprise paclitaxel, carboplatin, fluorouracil, oxaliplatin, cisplatin, irinotecan and/or capecitabine. The systemic therapy may, for example, comprise combination therapy with: paclitaxel and carboplatin; fluorouracil and oxaliplatin; fluorouracil and cisplatin; irinotecan and cisplatin; or paclitaxel and fluoropyrimidine (fluorouracil or capecitabine).

[0061] The systemic therapy may for example comprise a drug, or a combination of drugs, previously described for perioperative chemotherapy in a standard-of-care curative intent treatment treatment for operable gastroesophageal cancer (or gastroesophageal cancer for which local therapy is indicated). The gastroesophageal cancer may, for example, be adenocarcinoma of the thoracic esophagus or EGJ. The systemic therapy may, for example, comprise fluorouracil, leucovorin, oxaliplatin, docetaxel, fluoropyrimidine and/or cisplatin. The systemic therapy may, for example, comprise combination therapy with: fluorouracil, leucovorin, oxaliplatin, and docetaxel; fluoropyrimidine and oxaliplatin; or fluorouracil and cisplatin.

[0062] The systemic therapy may for example comprise a drug, or a combination of drugs, previously described for definitive chemoradiation in a standard-of-care curative intent treatment for operable gastroesophageal cancer (or gastroesophageal cancer for which local therapy is indicated). The systemic therapy may, for example, comprise paclitaxel, carboplatin, fluorouracil, oxaliplatin, cisplatin, docetaxel, irinotecan, and/or fluoropyrimidine (fluorouracil or capecitabine). The systemic therapy may, for example, comprise combination therapy with: paclitaxel and carboplatin; fluorouracil and oxaliplatin; fluorouracil and cisplatin; cisplatin with docetaxel or paclitaxel; irinotecan and cisplatin; or paclitaxel and fluoropyrimidine (fluorouracil or capecitabine).

[0063] The systemic therapy may for example comprise a drug, or a combination of drugs, previously described for postoperative therapy in a standard-of-care curative intent treatment treatment for operable gastroesophageal cancer (or gastroesophageal cancer for which local therapy is indicated). The postoperative therapy may, for example, comprise nivolumab, capecitabine, oxaliplatin, or fluorouracil. The systemic therapy may, for example, comprise monotherapy with nivolumab, for instance after postoperative therapy with resection. The systemic therapy may, for example, comprise combination therapy with: capecitabine and oxaliplatin; or fluorouracil and oxaliplatin.

[0064] The systemic therapy may for example comprise a drug, or a combination of drugs, previously described for postoperative chemoradiation in a standard-of-care curative intent treatment for operable gastroesophageal cancer (or gastroesophageal cancer for which local therapy is indicated). The systemic therapy may, for example, comprise fluoropyrimidine (i.e. fluorouracil or capecitabine). The systemic therapy may, for example, comprise monotherapy with fluorouracil or capecitabine, for instance before and/or after fluoropyrimidine-based chemoradiation.

[0065] In any case, the choice of systemic therapy for inclusion in a curative intent treatment may be informed by the nature of the cancer, such as its expression of particular markers. For example, if the cancer overexpresses HER2 (i.e. if the cancer is HER2 overexpression positive, for example a HER2 overexpression positive adenocarcinoma), the curative intent treatment may comprise trastuzumab (e.g. in combination with a fluoropyrimidine, oxaliplatin or cisplatin, and optionally pembrolizumab). HER2 overexpression may be determined by any means known to the skilled person, such as immunohistochemistry (IHC), fluorescence in-situ hybridisation (FISH), other in situ hybridisation (ISH) or next generation sequencing (NGS). In surgical specimens, a cancer may be considered positive for HER2 overexpression is there is strong complete, basolateral, or lateral membranous reactivity in 10% of cancer cells. In biopsy specimens, a cancer may be considered positive for HER2 overexpression is there is a cluster of five or more cancer cells with a strong complete, basolateral, or lateral membranous reactivity irrespective of percentage of cancer cells positive. HER2 overexpression is equivocal if the reactivity is weak to moderately complete.

[0066] If the cancer overexpresses PD-L1, the curative intent treatment may comprise a PD-1 axis binding antagonist, such as nivolumab or pembrolizumab. PD-L1 testing may be used to determine if an individual is a candidate for treatment with such an antagonist. Methods or determining PD-L1 expression are known to the skilled person. A cancer may be considered suitable for treatment with a PD-1 axis binding inhibitor if at least 1% of the gastroesophageal cancer cells from the individual may express PD-L1, such as at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70% or at least 80%. The percentage of cells that express PD-L1 may be determined by any means known to the skilled person, such as immunohistochemistry (IHC), flow-cytometry or enzyme-linked immunosorbent assay (ELISA). A cancer may be considered to have PD-L1 expression if its combined positive score (CPS) is 1, for example, 5 or 10.

[0067] If the cancer is an NTRK gene fusion-positive solid tumour, the curative intent treatment may comprise a TRK inhibitor such as larotrectinib or entrectinib. Said tumours may be identified by any means known to the skilled person, such as with next generation sequencing (NGS). Tumours with a high mutational burden (TMB) may be determined by the skilled person using NGS.

[0068] In one aspect of the disclosure, the curative intent treatment does not comprise therapeutic T cells. Accordingly, the individual may not have received therapeutic T cells prior to the method of the disclosure.

First-Line and Second-Line Treatments

[0069] The first-line and/or second-line treatment may be any known or unknown treatment for gastroesophageal cancer. As explained above, known treatments for gastroesophageal cancer include surgical resection, radiation therapy and systemic therapy. A curative intent treatment (such as the first curative intent treatment or the second curative intent treatment) may therefore comprise (i) surgical resection, (ii) radiation therapy, and/or (iii) systemic therapy. For example, a curative intent treatment may comprise (i); (ii); (iii); (i) and (ii); (i) and (iii); (ii) and (iii); or (i), (ii) and (iii). In particular aspects of the disclosure, a curative intent treatment may comprise (i); (ii); or (i) and (ii).

[0070] The systemic therapy may comprise or consist of (a) a chemotherapy. The systemic therapy may comprise or consist of (b) an immunotherapy. The systemic therapy may comprise or consist of (c) a targeted therapy. For example, the systemic therapy may comprise or consist of (a); (b); (c); (a) and (b); (a) and (c); (b) and (c); or (a), (b) and (c).

[0071] Chemotherapies, such as those for gastroesophageal cancer, are well-known in the art. Such chemotherapies may, for example, comprise a platinum-based anti-neoplastic drug, such as cisplatin, oxaliplatin or carboplatin. Such chemotherapies may, for example, comprise an anti-metabolite, such as fluorouracil (5-FU), trifluridine or capecitabine. Such chemotherapies may, for example, comprise a taxane drug, such as docetaxel or paclitaxel. Leucovorin may be administered with a chemotherapy to reduce the toxic effects of the chemotherapy.

[0072] Immunotherapies, such as those for gastroesophageal cancer, are well-known in the art. Such immunotherapies may, for example, include therapeutic immune cells, immunomodulators, checkpoint inhibitors, and vaccines. Therapeutic immune cells may include T cells, for instance engineered T cells such as CAR T cells or T cells expressing an engineered TCR. Immunomodulators may include, for example, interleukins, cytokines, chemokines, and immunomodulatory imide drugs. Checkpoint inhibitors may, for instance, include CTLA-4 inhibitors or PD-1 axis binding antagonists. CTLA-4 inhibitors may include ipilimumab. PD-1 axis binding antagonists may, for instance, include pembrolizumab, dostarlimab-gxly and nivolumab. Checkpoint inhibitors and PD-1 axis binding antagonists are described in detail below.

[0073] Targeted therapies, such as those for gastroesophageal cancer, are well-known in the art. The term targeted therapy is a term of art that refers to treatments that target specific genes and proteins that help cancer cells survive and grow. Targeted therapies may, for example, include a tropomysin kinase receptor antagonist (such as larotrectinib or entrectinib), a HER2 antagonist (such as trastuzumab), a topoisomerase I inhibitor (such as irinotecan or deruxtecan-nxki), a VEGF inhibitor (such as ramucirumab), or a thymidine phosphorylase inhibitor (such as tiparicil).

[0074] The systemic therapy may, for example, comprise a drug (or a combination of drugs) previously described for therapy of gastroesophageal cancer. The systemic therapy may comprise a drug, or a combination of drugs, previously described as a first-line approved or standard-of-care treatment for gastroesophageal cancer.

[0075] For example, the systemic therapy may comprise a drug (or a combination of drugs) previously described for standard-of-care first-line therapy of gastroesophageal cancer, such as inoperable gastroesophageal cancer (or gastroesophageal cancer where surgical approaches are less preferred). The systemic therapy may, for example, comprise fluoropyrimidine (fluorouracil or capecitabine), oxaliplatin, trastuzumab, cisplatin, nivolumab, pembrolizumab, ipilimumab, irinotecan, paclitaxel, carboplatin and/or docetaxel. The systemic therapy may, for example, comprise combination therapy with: fluoropyrimidine (fluorouracil or capecitabine) and oxaliplatin and trastuzumab (e.g. for HER2 overexpression positive adenocarcinoma); fluoropyrimidine (fluorouracil or capecitabine) and cisplatin and trastuzumab (e.g. for HER2 overexpression positive adenocarcinoma); fluoropyrimidine (fluorouracil or capecitabine), oxaliplatin, and nivolumab for adenocarcinoma (e.g. for HER2 overexpression negative adenocarcinoma); fluoropyrimidine (fluorouracil or capecitabine), oxaliplatin, and nivolumab (e.g. for HER2 overexpression negative squamous cell carcinoma); fluoropyrimidine (fluorouracil or capecitabine), cisplatin, and nivolumab (e.g. for HER2 overexpression negative squamous cell carcinoma); fluoropyrimidine (fluorouracil or capecitabine), oxaliplatin, and pembrolizumab (e.g. for HER2 overexpression negative cancers); fluoropyrimidine (fluorouracil or capecitabine), cisplatin, and pembrolizumab (e.g. for HER2 overexpression negative cancers); fluoropyrimidine (fluorouracil or capecitabine) and oxaliplatin (e.g. for HER2 overexpression negative cancers); fluoropyrimidine (fluorouracil or capecitabine) and cisplatin (e.g. for HER2 overexpression negative cancers); nivolumab and ipilimumab (e.g. for HER2 overexpression negative squamous cell carcinoma); fluoropyrimidine (fluorouracil or capecitabine) and cisplatin and trastuzumab and pembrolizumab (e.g. for HER2 overexpression positive adenocarcinoma); fluoropyrimidine (fluorouracil or capecitabine) and oxaliplatin and trastuzumab and pembrolizumab (e.g. for HER2 overexpression positive adenocarcinoma); fluorouracil and irinotecan; paclitaxel with or without cisplatin or carboplatin; docetaxel with or without cisplatin; fluoropyrimidine (fluorouracil or capecitabine); docetaxel, cisplatin or oxaliplatin, and fluorouracil; or docetaxel, carboplatin, and fluorouracil.

[0076] The systemic therapy may, for example, comprise a drug (or a combination of drugs) previously described for approved or standard-of-care second-line or subsequent-line therapy of gastroesophageal cancer, such as inoperable gastroesophageal cancer (or gastroesophageal cancer where surgical approaches are less preferred). The systemic therapy may, for example, comprise dostarlimab-gxly, nivolumab, pembrolizumab, docetaxel, paclitaxel, irinotecan, entrectinib, larotrectinib, ramucirumab, fam-trastuzumab, deruxtecan-nxki, fluorouracil, and/or cisplatin. The systemic therapy may, for example, comprise monotherapy with dostarlimab-gxly (e.g. for MSI-H or dMMR tumours), nivolumab (e.g. for esophageal squamous cell carcinoma), pembrolizumab (e.g. for MSI-H or dMMR tumours, or for TMB high (10 mutations/megabase) tumours, or for second-line therapy for esophageal squamous cell carcinoma with PD-L1 expression levels by CPS of 10), docetaxel, paclitaxel, irinotecan, entrectinib (e.g. for NTRK gene fusion-positive tumours) or larotrectinib (e.g. for NTRK gene fusion-positive tumours). The systemic therapy may, for example, comprise combination therapy with: ramucirumab and paclitaxel (e.g. for adenocarcinoma such as EGJ adenocarcinoma or esophageal adenocarcinoma); fam-trastuzumab deruxtecan-nxki (e.g. for HER2 overexpression positive adenocarcinoma); fluorouracil and irinotecan; ramucirumab for adenocarcinoma (e.g. for EGJ adenocarcinoma or esophageal adenocarcinoma); irinotecan and cisplatin; fluorouracil and irinotecan and ramucirumab (e.g. for adenocarcinoma); irinotecan and ramucirumab (e.g. for adenocarcinoma); or docetaxel and irinotecan.

[0077] The systemic therapy may, for example, comprise a drug (or a combination of drugs) previously described for approved or standard-of-care third-line or subsequent-line therapy of gastroesophageal cancer, such as inoperable gastroesophageal cancer (or gastroesophageal cancer where surgical approaches are less preferred). The systemic therapy may, for example, comprise trifluridine and/or tipiracil. The systemic therapy may, for example, comprise combination therapy with trifluridine and tipiracil (e.g. for EGJ adenocarcinoma).

[0078] In any case, the choice of systemic therapy for inclusion in a treatment (such as the first-line or second-line treatment) may be informed by the nature of the cancer, such as its expression of particular markers. For example, if the cancer overexpresses HER2 (i.e. if the cancer is HER2 overexpression positive, for example a HER2 overexpression positive adenocarcinoma), the first-line and/or second-line treatment may comprise trastuzumab (e.g. in combination with a fluoropyrimidine, oxaliplatin or cisplatin, and optionally pembrolizumab). HER2 overexpression may be determined by any means known to the skilled person, such as immunohistochemistry (IHC), fluorescence in-situ hybridisation (FISH), other in situ hybridisation (ISH) or next generation sequencing (NGS). In surgical specimens, a cancer may be considered positive for HER2 overexpression is there is strong complete, basolateral, or lateral membranous reactivity in 10% of cancer cells. In biopsy specimens, a cancer may be considered positive for HER2 overexpression is there is a cluster of five or more cancer cells with a strong complete, basolateral, or lateral membranous reactivity irrespective of percentage of cancer cells positive. HER2 overexpression is equivocal if the reactivity is weak to moderately complete.

[0079] If the cancer overexpresses PD-L1, the first-line and/or second-line treatment may comprise a PD-1 axis binding antagonist, such as nivolumab or pembrolizumab. PD-L1 testing may be used to determine if an individual is a candidate for treatment with such an antagonist. Methods or determining PD-L1 expression are known to the skilled person. A cancer may be considered suitable for treatment with a PD-1 axis binding inhibitor if at least 1% of the gastroesophageal cancer cells from the individual may express PD-L1, such as at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70% or at least 80%. The percentage of cells that express PD-L1 may be determined by any means known to the skilled person, such as immunohistochemistry (IHC), flow-cytometry or enzyme-linked immunosorbent assay (ELISA). A cancer may be considered to have PD-L1 expression if its combined positive score (CPS) is 1, for example, 5 or 10.

[0080] If the cancer is an NTRK gene fusion-positive solid tumour, the first-line and/or second-line treatment may comprise a TRK inhibitor such as larotrectinib or entrectinib. Said tumours may be identified by any means known to the skilled person, such as with next generation sequencing (NGS). Tumours with a high mutational burden (TMB) may be determined by the skilled person using NGS.

[0081] In one aspect of the disclosure, the first-line and/or second-line treatment does not comprise therapeutic T cells. Accordingly, the individual may not have received therapeutic T cells prior to the method of the disclosure.

Population of Modified T Cells

[0082] The method comprises administering to the individual a population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous TCR capable of binding to a peptide antigen of MAGE-A4. It is the presence of the heterologous CD8 co-receptor and a heterologous TCR that renders the T cells modified. The heterologous CD8 co-receptor and the heterologous TCR are typically present on the surface of the modified T cells. In other words, the modified T cells may express the heterologous CD8 co-receptor and the heterologous TCR on their surface.

[0083] In the context of the present disclosure, the term heterologous refers to a polypeptide or nucleic acid that is foreign to a particular biological system (such as a T cell), i.e. that is not naturally present in that system. A heterologous polypeptide or nucleic acid may be introduced to the system by artificial or recombinant means. Accordingly, heterologous expression of a TCR may alter the specificity of a T cell. Heterologous expression of a CD8 co-receptor may endow the T cell with functions associated with the CD8 co-receptor. The heterologous CD8 co-receptor and the heterologous TCR are described in detail below.

[0084] The modified T cells may comprise CD4+ T cells. That is, the modified T cells may comprise T cells expressing an endogenous CD4 co-receptor. The modified T cells may comprise CD8+ T cells. That is, the modified T cells may comprise T cells expressing an endogenous CD8 co-receptor. The modified T cells may comprise CD4+ T cells and CD8+ T cells. That is, the modified T cells may comprise T cells expressing an endogenous CD4 co-receptor, and comprise T cells expressing an endogenous CD8 co-receptor. Both CD4+ T cells and CD8+ T cells are capable of harbouring a heterologous CD8 co-receptor.

[0085] The modified T cells may be allogeneic with respect to the individual. The modified T cells may preferably be autologous with respect to the individual. In this case, the modified T cells may be produced by modifying endogenous cells obtained from the individual. Thus, the method may comprise producing the population. Methods for producing modified T cells are known in the art and considered in the Example below. Typically, the modified T cells of the disclosure are produced from cells, such as peripheral blood mononuclear cells (PBMCs). T cells are typically selected from the harvested cells, and manipulated to comprise the desired modifications (here, the heterologous CD8 co-receptor and the heterologous TCR). The method may therefore comprise producing the population by: (a) obtaining peripheral blood mononuclear cells (PBMCs) from the individual; (b) selecting T cells from the PBMCs; and (c) modifying the selected T cells to express the heterologous CD8 co-receptor and the heterologous TCR.

[0086] Autologous modified T cells may be produced in anticipation of the individual's need for them. That is, autologous modified T cells may be produced ahead of time, before an individual requires treatment with the modified cells. This can help to ensure that autologous modified T cells are available for administration as soon as possible after the individual is identified as requiring treatment. In this way, the individual need not wait for autologous T cells to be produced before treatment can begin. This may improve the outcome of treatment.

[0087] Production of autologous T cells ahead of time may be particularly relevant in the treatment of gastroesophageal cancers with a high risk of relapse and/or with a high risk of failure of the first-line and/or second-line standard-of-care treatment. The risk of relapse in an individual subjected to standard-of-care treatment for gastroesophageal cancer may be determined by methods routine in the art. Such methods may include monitoring for clinical signs or symptoms. Such methods may include, for example, one or more magnetic resonance imaging (MRI), positron emission tomography (PET) and/or computerised tomography (CT) scans conducted following treatment, to monitor for progression and/or return of tumours. MRI, PET and/or CT scans may, for example, be performed about every three months (such as once every 4 to 16 weeks, or once every 8 to 12 weeks). If an individual is identified as high risk of relapse, PBMCs may be obtained at this point (i.e. prior to relapse) with a view to producing autologous modified T cells ready for administration when relapse occurs. Accordingly, when the cancer is relapsed gastroesophageal cancer, the method of the disclosure may comprise producing the population by (a) obtaining peripheral blood mononuclear cells (PBMCs) from the individual; (b) selecting T cells from the PBMCs; and (c) modifying the selected T cells to express the heterologous CD8 co-receptor and the heterologous TCR, wherein one or more of steps (a) to (c) are performed prior to relapse. Preferably, step (a) is performed prior to relapse. More preferably, steps (a) and (b) are performed prior to relapse. Most preferably, steps (a), (b) and (c) are performed prior to relapse. Any of these options effectively gives treatment a head-start on relapse.

[0088] In any case, the population of modified T cells may, for example, be administered as a single dose. The population of modified T cells may, for example, be administered as soon as possible after diagnosis of the gastroesophageal cancer, for instance as a single dose. For example, the population of modified T cells may be administered as soon as possible after relapse of gastroesophageal cancer is identified. The population of modified T cells may, for example, be administered as soon as possible after previously-untreated gastroesophageal cancer is identified.

[0089] In the context of the present disclosure, the term as soon as possible may refer to the earliest point that it is practical to administer the population of modified T cells. As explained above, production of autologous modified T cells may take some time, potentially leading to a lag between diagnosis of the gastroesophageal cancer gastroesophageal cancer (e.g. relapsed gastroesophageal cancer) and the implementation of therapy. Therefore, administration as soon as possible after diagnosis may refer to administration as soon as is practical once autologous modified T cells have been produced. Administration as soon as possible after diagnosis may, for example, refer to administration from less than about 150 days after diagnosis of the gastroesophageal cancer (e.g. relapsed gastroesophageal cancer), such as less than about 125 days, less than about 100 days, less than about 90 days, less than about 80 days, less than about 70 days, less than about 60 days, less than about 50 days, less than about 40 days, or less than about 30 days after diagnosis of the gastroesophageal cancer (e.g. relapsed gastroesophageal cancer). The population may be administered to the individual, for example, about 30 to about 150 days after diagnosis of the gastroesophageal cancer (e.g. relapsed gastroesophageal cancer), such as about 40 to about 125, about 50 to about 100, about 90, about 85, about 80, about 75, about 70, about 65 or about 60 days after diagnosis of the gastroesophageal cancer (e.g. relapsed gastroesophageal cancer).

Heterologous TCR

[0090] The modified T cells comprise a heterologous TCR capable of binding to a peptide antigen of MAGE-A4. In other words, the modified T cells express or present a heterologous TCR capable of binding to a peptide antigen of MAGE-A4, for instance on their surface. MAGE-A4 is a well-known cancer antigen that has restricted expression in normal (i.e. non-cancerous) tissue. MAGE-A4 has been shown to repress p53 targets (such as BAX and CDKN1A) and is a binding partner for the oncogene gankyrin.

[0091] The heterologous TCR is capable of binding to a peptide antigen of MAGE-A4. The heterologous TCR may, for example, bind to GVYDGREHTV (SEQ ID NO: 1) which is a peptide sequence known as MAGE-A4 230-239 that is comprised in MAGE-A4. The heterologous TCR may, for example, bind to a complex comprising a peptide antigen of MAGE-A4 (e.g. GVYDGREHTV (SEQ ID NO: 1)) and an HLA molecule, such as an HLA-A molecule (e.g. an HLA-A*02 or an HLA-A*0201 molecule). In any case, the binding may be specific. Specificity refers to the strength of binding between the heterologous TCR and its target antigen. Specificity may be described by a dissociation constant, Kd, the ratio between bound and unbound states for the receptor-ligand system. Typically, the fewer different antigens the heterologous TCR is capable of binding other than MAGE-A4, the greater its binding specificity.

[0092] The heterologous TCR may, for example, bind to a peptide antigen of MAGE-A4 (e.g. SEQ ID NO: 1), or to a complex comprising a peptide antigen of MAGE-A4 (e.g. SEQ ID NO: 1) and an HLA molecule, with a dissociation constant (Kd) of between 0.01 M and 100 M, between 0.01 M and 50 M, between 0.01 M and 20 M, between 10 M and 1000 M, between 10 M and 500 M, or between 50 M and 500 M. For instance, in a preferred aspect of the disclosure, the heterologous TCR binds to a peptide antigen of MAGE-A4, or to a complex comprising a peptide antigen of MAGE-A4 and an HLA molecule, with a Kd of between 0.05 M to 20.0 M. For example, the heterologous TCR may bind to a peptide antigen of MAGE-A4, or to a complex comprising a peptide antigen of MAGE-A4 and an HLA molecule, with a Kd of 0.01 M, 0.02 M, 0.03 M, 0.04 M, 0.05 M, 0.06 M, 0.07 M, 0.08 M, 0.09 M, 0.1 M, 0.15 M, 0.2 M, 0.25 M, 0.3 M, 0.35 M, 0.4 M, 0.45 M, 0.5 M, 0.55 M, 0.6 M, 0.65 M, 0.7 M, 0.75 M, 0.8 M, 0.85 M, 0.9 M, 0.95 M, 1.0 M, 1.5 M, 2.0 M, 2.5 M, 3.0 M, 3.5 M, 4.0 M, 4.5 M, 5.0 M, 5.5 M, 6.0 M, 6.5 M, 7.0 M, 7.5 M, 8.0 M, 8. M, 9.0 M, 9.5 M, 10.0 M, 20 M, 30 M, 40 M, 50 M, 60 M, 70 M, 80 M, 90 M, 100 M, 150 M, 200 M, 250 M, 300 M, 350 M, 400 M, 450 M, 500 M. The Kd may, for example, be measured using surface plasmon resonance, optionally at 25 C., optionally between a pH of 6.5 and 6.9 or 7.0 and 7.5. The dissociation constant, Kd or koff/kon may be determined by experimentally measuring the dissociation rate constant, koff, and the association rate constant, kon. A TCR dissociation constant may be measured using a soluble form of the TCR, wherein the TCR comprises a TCR alpha chain variable domain and a TCR beta chain variable domain.

[0093] The heterologous TCR may, for example, be a recombinant or synthetic or artificial TCR. That is, the heterologous TCR may be a TCR that does not exist in nature. The heterologous TCR may, for example, be an affinity enhanced TCR, for example a specific peptide enhanced affinity receptor (SPEAR) TCR.

[0094] The heterologous TCR may, for example, comprise an alpha chain variable domain having at least 80% sequence identity to the sequence of amino acid residues 22-125 of SEQ ID NO: 2. The heterologous TCR may, for example, comprise a beta chain variable domain having at least 80% sequence identity to the sequence of amino acid residues 22-123 of SEQ ID NO: 3. The heterologous TCR may, for example, comprise an alpha chain variable domain having at least 80% sequence identity to the sequence of amino acid residues 22-125 of SEQ ID NO: 2 and a beta chain variable domain having at least 80% sequence identity to the sequence of amino acid residues 22-123 of SEQ ID NO: 3. The alpha chain variable domain may, for example, have at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% sequence identity to the sequence of amino acid residues 22-125 of SEQ ID NO: 2. The alpha chain variable domain may, for example, comprise or consist of amino acid residues 22-125 of SEQ ID NO: 2. The beta chain variable domain may, for example, have at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% sequence identity to the sequence of amino acid residues 22-123 of SEQ ID NO: 3. The beta chain amino acid sequence may, for example, comprise or consist of amino acid residues 22-123 of SEQ ID NO: 3.

[0095] The heterologous TCR may, for example, comprise an alpha chain having at least 80% sequence identity to the sequence of amino acid residues 22-282 of SEQ ID NO: 2. The heterologous TCR may, for example, comprise a beta chain having at least 80% sequence identity to the sequence of amino acid residues 22-311 of SEQ ID NO: 3. The heterologous TCR may, for example, comprise an alpha chain having at least 80% sequence identity to the sequence of amino acid residues 22-282 of SEQ ID NO: 2 and a beta chain variable domain having at least 80% sequence identity to the sequence of amino acid residues 22-311 of SEQ ID NO: 3. The alpha chain variable domain may, for example, have at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% sequence identity to the sequence of amino acid residues 22-282 of SEQ ID NO: 2. The alpha chain variable domain may, for example, comprise or consist of amino acid residues 22-282 of SEQ ID NO: 2. The beta chain variable domain may, for example, have at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% sequence identity to the sequence of amino acid residues 22-311 of SEQ ID NO: 3. The beta chain amino acid sequence may, for example, comprise or consist of amino acid residues 22-311 of SEQ ID NO: 3.

[0096] The heterologous TCR is typically expressed with N-terminal signal peptides that are cleaved prior to expression at the surface of the T cell. In this respect, amino acids 1 to 21 of each of SEQ ID NO: 2 and SEQ ID NO: 3 are typically cleaved prior to expression of the TCR at the surface of the T cell. The heterologous TCR may, for example, comprise an alpha chain amino acid sequence having at least 80% sequence identity to SEQ ID NO: 2. The heterologous TCR may, for example, comprise a beta chain amino acid sequence having at least 80% sequence identity to SEQ ID NO: 3. The heterologous TCR may, for example, comprise an alpha chain amino acid sequence having at least 80% sequence identity to SEQ ID NO: 2 and a beta chain amino acid sequence having at least 80% sequence identity to SEQ ID NO: 3. The alpha chain amino acid sequence may, for example, have at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO: 2. The alpha chain amino acid sequence may, for example, comprise or consist of SEQ ID NO: 2. The beta chain amino acid sequence may, for example, have at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO: 3. The beta chain amino acid sequence may, for example, comprise or consist of SEQ ID NO: 3.

[0097] The heterologous TCR may, for example, comprise (i) an alpha chain variable domain that comprises a CDR1 that comprises (1) the sequence of SEQ ID NO: 4 or (2) an amino acid sequence that comprises one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 4; (ii) an alpha chain variable domain that comprises a CDR2 that comprises (1) the sequence of SEQ ID NO: 5 or (2) an amino acid sequence that comprises one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 5; (iii) an alpha chain variable domain that comprises a CDR3 that comprises (1) the sequence of SEQ ID NO: 6 or (2) an amino acid sequence that comprises one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 6; (iv) a beta chain variable domain that comprises a CDR1 that comprises (1) the sequence of SEQ ID NO: 7 or (2) an amino acid sequence that comprises one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO:7; (v) a beta chain variable domain that comprises a CDR2 that comprises (1) the sequence of SEQ ID NO: 8 or (2) an amino acid sequence that comprises one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 8; and/or (vi) a beta chain variable domain that comprises a CDR3 that comprises (1) the sequence of SEQ ID NO: 9 or (2) an amino acid sequence that comprises one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 9.

[0098] The alpha chain of the heterologous TCR may, for example, comprise (i) an alpha chain variable domain that comprises a CDR1 that comprises (1) the sequence of SEQ ID NO: 4 or (2) an amino acid sequence that comprises one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 4; (ii) an alpha chain variable domain that comprises a CDR2 that comprises (1) the sequence of SEQ ID NO: 5 or (2) an amino acid sequence that comprises one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 5; and (iii) an alpha chain variable domain that comprises a CDR3 that comprises (1) the sequence of SEQ ID NO: 6 or (2) an amino acid sequence that comprises one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 6. The alpha chain of the heterologous TCR may, for example, comprise (i) an alpha chain variable domain that comprises a CDR1 that comprises the sequence of SEQ ID NO: 4; (ii) an alpha chain variable domain that comprises a CDR2 that comprises the sequence of SEQ ID NO: 5; and (iii) an alpha chain variable domain that comprises a CDR3 that comprises the sequence of SEQ ID NO: 6.

[0099] The beta chain of the heterologous TCR may, for example, comprise (iv) a beta chain variable domain that comprises a CDR1 that comprises (1) the sequence of SEQ ID NO: 7 or (2) an amino acid sequence that comprises one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO:7; (v) a beta chain variable domain that comprises a CDR2 that comprises (1) the sequence of SEQ ID NO: 8 or (2) an amino acid sequence that comprises one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 8; and (vi) a beta chain variable domain that comprises a CDR3 that comprises (1) the sequence of SEQ ID NO: 9 or (2) an amino acid sequence that comprises one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 9. The beta chain of the heterologous TCR may, for example, comprise (iv) a beta chain variable domain that comprises a CDR1 that comprises the sequence of SEQ ID NO: 7; (v) a beta chain variable domain that comprises a CDR2 that comprises the sequence of SEQ ID NO: 8; and (vi) a beta chain variable domain that comprises a CDR3 that comprises the sequence of SEQ ID NO: 9.

[0100] The heterologous TCR may, for example, comprise an alpha chain comprising a CDR1 having the sequence of SEQ ID NO: 4, a CDR2 having the sequence of SEQ ID NO: 5 and a CDR3 having the sequence of SEQ ID NO: 6, and a beta chain comprising a CDR1 having the sequence of SEQ ID NO: 7, a CDR2 having the sequence of SEQ ID NO: 8 and a CDR3 having the sequence of SEQ ID NO: 9. The heterologous TCR may, for example, have additionally any of the percentage identities in the alpha chain and beta chain discussed herein.

Heterologous CD8 Co-Receptor

[0101] The modified T cells comprise a heterologous CD8 co-receptor. In other words, the modified T cells express a heterologous CD8 co-receptor, for instance on their surface.

[0102] CD8 is a cell surface glycoprotein that, in nature, is found on most cytotoxic T lymphocytes and mediates efficient cell-cell interactions within the immune system. CD8 acts as a co-receptor for the T cell receptor, such that CD8 and the T cell receptor together recognise antigen displayed by an antigen-presenting cell in the context of class I MHC molecules. The CD8 co-receptor binds to class 1 MHCs and potentiates TCR signaling. The functional co-receptor may be a homodimer consisting of two CD8 alpha chains, or a heterodimer consisting of one CD8 alpha chain and one CD8 beta chain.

[0103] Accordingly, the heterologous CD8 co-receptor comprised in the modified T cells may be CD8. In other words, the heterologous CD8 co-receptor may be a homodimer consisting of two CD8 alpha chains. Alternatively, the heterologous CD8 co-receptor may be a heterodimer consisting of one CD8 alpha chain and one CD8 beta chain. In either case, a CD8 alpha chain may comprise or consist of an amino acid sequence having at least 80% (such as at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99% or 100%) sequence identity to SEQ ID NO: 10. Thus, the heterologous CD8 co-receptor may comprise an amino acid sequence having at least 80% (such as at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99% or 100%) sequence identity to SEQ ID NO: 10.

[0104] CD8 alpha chains and CD8 beta chains both share significant homology to immunoglobulin variable light chains. CD8 alpha chains and beta chains have CDR-like loops involved in MHC-Class I binding. The heterologous CD8 co-receptor may, for example, comprise a CD8 alpha chain that comprises: (i) an alpha chain CDR1 that comprises (1) the sequence of SEQ ID NO: 11 or (2) an amino acid sequence that comprises one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 11; (ii) an alpha chain CDR2 that comprises (1) the sequence of SEQ ID NO: 12 or (2) an amino acid sequence that comprises one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 12; and/or (iii) an alpha chain CDR3 that comprises (1) the sequence of SEQ ID NO: 13 or (2) an amino acid sequence that comprises one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 13.

[0105] The heterologous CD8 co-receptor is capable of binding to a class I MHC molecule. The heterologous CD8 co-receptor may, for example, bind to the a3 portion of a class I MHC molecule, for instance via the IgV-like domain of the CD8 co-receptor. The a3 portion is typically found between residues 223 and 229 of a class I MHC molecule. The ability of the heterologous CD8 co-receptor to bind to a class I MHC molecule improves the ability of the modified T cells to engage cognate antigen via their heterologous TCR. The cognate antigen, MAGE-A4, is typically presented in complex with a class I MHC molecule such as HLA-A*02. The heterologous CD8 co-receptor may improve or increase the off-rate (k.sub.off) of the TCR/peptide-MHCI interaction in the modified cells. The improvement or increase may be relative to modified T cells that comprise a heterologous TCR that binds to MAGE-A4 but which lack a heterologous CD8 co-receptor. The heterologous CD8 co-receptor may, for example, assist in organising the heterologous TCR on the surface of modified cells, thereby improving the ability of the heterologous TCR to participate in the TCR/peptide-MHCI interaction. The heterologous CD8 co-receptor may, for example, bind or interact with LCK (lymphocyte-specific protein tyrosine kinase) in a zinc-dependent manner leading to activation of transcription factors like NFAT, NF-B, and AP-1. Accordingly, expression of a heterologous CD8 co-receptor may confer upon the modified T cells an improved affinity and/or avidity for MAGE-A4, and/or improved activation upon binding to MAGE-A4. Methods for determining affinity, avidity and T cell activation are well-known in the art. Expression of a heterologous CD8 co-receptor may confer upon the modified T cells an improved or increased expression of CD40L, cytokine production, cytotoxic activity, induction of dendritic cell maturation or induction of dendritic cell cytokine production, for instance in response to antigen (MAGE-A4) binding. Improvements or increases may be relative to modified T cells that comprise a heterologous TCR that binds to a peptide antigen of MAGE-A4 but which lack a heterologous CD8 co-receptor.

[0106] Synergy has been demonstrated between CD8a and peptide antigen presented on HLA-A*0201. Therefore, in one aspect of the disclosure, the heterologous CD8 co-receptor may be CD8a and the heterologous TCR may be capable of binding to a peptide antigen of MAGE-A4 in complex with HLA-A*0201. The peptide antigen may, for example, be SEQ ID NO: 1.

Additional Anti-Cancer Therapy

[0107] The method may further comprise administering an additional anti-cancer therapy to the individual. That is, the method may comprise administering to the individual (i) a population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous T-cell receptor (TCR) capable of binding to a peptide antigen of MAGE-A4, and (ii) an additional anti-cancer therapy. In other words, the method may comprise combination treatment with (i) a population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous T-cell receptor (TCR) capable of binding to a peptide antigen of MAGE-A4, and (ii) an additional anti-cancer therapy. A checkpoint inhibitor, such as a PD-1 axis binding antagonist, may also be included in the combination as set out below.

[0108] At least one additional anti-cancer therapy may be administered to the individual. For example, one or more, two or more, three or more, four or more, or five or more additional anti-cancer therapies may be administered to the individual.

[0109] The additional anti-cancer therapy may be administered to the individual in the same line of therapy as the population of modified T cells. As set out above, a line of therapy may refer to a particular treatment regimen for a cancer for which curative intent treatment has failed, or for which curative intent treatment may be inappropriate. The additional anti-cancer therapy and the population of modified T cells may be administered as part of a first-line treatment regimen. The additional anti-cancer therapy and the population of modified T cells may be administered as part of a second-line treatment regimen. The second-line treatment regimen may, for example, be employed following failure of the first-line treatment regimen. The additional anti-cancer therapy and the population of modified T cells may be administered as part of a third-line treatment regimen. The third-line treatment regimen may, for example, be employed following failure of the second-line treatment regimen.

[0110] The additional anti-cancer therapy may, for example, be an additional anti-cancer drug therapy. In other words, the additional anti-cancer therapy may be a systemic therapy. The additional anti-cancer therapy may, for example, comprise or consist of (a) a chemotherapy. The additional anti-cancer therapy may, for example, comprise or consist of (b) an immunotherapy. The additional anti-cancer therapy may, for example, comprise or consist of (c) a targeted therapy. For example, the additional anti-cancer therapy may comprise or consist of (a); (b); (c); (a) and (b); (a) and (c); (b) and (c); or (a), (b) and (c).

[0111] Any chemotherapy may be comprised in the additional anti-cancer therapy. The method may comprise administering one or more chemotherapeutic agents (such as two or more, three or more, four or more, or five or more chemotherapeutic agents). Chemotherapies, such as those for gastroesophageal cancer, are well-known in the art. Such chemotherapies may, for example, comprise a platinum-based anti-neoplastic drug, such as cisplatin, oxaliplatin or carboplatin. Such chemotherapies may, for example, comprise an anti-metabolite, such as a fluoropyrimidine (e.g. fluorouracil or capecitabine), gemcitabine, trifluridine or methotrexate. Such chemotherapies may, for example, comprise administering a taxane drug, such as docetaxel or paclitaxel. Such chemotherapies may, for example, comprise a topoisomerase I inhibitor, such as irinotecan or deruxtecan-nxki. Such chemotherapies may, for example, comprise a thymidine phosphorylase inhibitor, such as tipiracil. Leucovorin may be administered with the one or more chemotherapeutic agents to minimise toxic effects.

[0112] Any immunotherapy may be comprised in the additional anti-cancer therapy. The method may comprise administering one or more immunotherapies (such as two or more, three or more, four or more, or five or immunotherapies). Immunotherapies, such as those for gastroesophageal cancer, are well-known in the art. Such immunotherapies may, for example, include therapeutic immune cells, immunomodulators, checkpoint inhibitors, and vaccines. Therapeutic immune cells may include T cells, for instance engineered T cells such as CAR T cells or T cells expressing an engineered TCR. Immunomodulators may include, for example, interleukins, cytokines, chemokines, and immunomodulatory imide drugs. Checkpoint inhibitors may, for instance, include CTLA-4 inhibitors or PD-1 axis binding antagonists. CTLA-4 inhibitors may include ipilimumab. PD-1 axis binding antagonists may, for instance, include pembrolizumab, dostarlimab-gxly and nivolumab. Checkpoint inhibitors and PD-1 axis binding antagonists are described in detail below. Any targeted therapy may be comprised in the additional anti-cancer therapy. The method may comprise administering one or more targeted therapies (such as two or more, three or more, four or more, or five or more targeted therapies). Targeted therapies, such as those for gastroesophageal cancer, are well-known in the art. Such targeted therapies may, for example, comprise ramucirumab, which is a direct VEGFR2 antagonist. Such targeted therapies may, for example, comprise entrectinib, which is a selective tyrosine kinase inhibitor, of the tropomyosin receptor kinases A, B and C, C-ros oncogene 1 and anaplastic lymphoma kinase. Such targeted therapies may, for example, comprise larotrectinib, which is an inhibitor of tropomyosin kinase receptors TrkA, TrkB, and TrkC. Such targeted therapies may, for example, comprise trastuzumab, a HER2-specific antibody. Other targeted therapies include bemarituzumab (FPA144), DKN-01, tebotelimab (MGD013), vactosertib (TEW-7197), zolbetuximab (IMAB362).

[0113] The additional anti-cancer therapy may comprise a drug, or a combination of drugs, previously described for therapy of gastroesophageal cancer. The additional anti-cancer therapy may comprise a drug, or a combination of drugs, previously described as a standard-of-care treatment for gastroesophageal cancer. Accordingly, the method may comprise administering a drug, or a combination of drugs, previously described for therapy of gastroesophageal cancer. The method may comprise administering a drug, or a combination of drugs, previously described as an approved standard-of-care treatment for gastroesophageal cancer. In this context, approval may relate to approval by the FDA, EMA or MHRA for example.

[0114] The additional anti-cancer therapy may, for example, comprise a drug (or a combination of drugs) previously described for use in preoperative chemoradiation for gastroesophageal cancer. The additional anti-cancer therapy may comprise a drug, or a combination of drugs, previously described as a standard-of-care preoperative chemoradiation treatment for gastroesophageal cancer. The gastroesophageal cancer may, for example, be an operable gastroesophageal cancer, or a gastroesophageal cancer for which local therapy is indicated. The additional anti-cancer therapy may, for example, comprise paclitaxel, carboplatin, fluorouracil, oxaliplatin, cisplatin, irinotecan and/or capecitabine. The additional anti-cancer therapy may, for example, comprise combination therapy with: paclitaxel and carboplatin; fluorouracil and oxaliplatin; fluorouracil and cisplatin; irinotecan and cisplatin; or paclitaxel and fluoropyrimidine (fluorouracil or capecitabine).

[0115] The additional anti-cancer therapy may, for example, comprise a drug (or a combination of drugs) previously described for use in perioperative chemotherapy for gastroesophageal cancer. The additional anti-cancer therapy may comprise a drug, or a combination of drugs, previously described as a standard-of-care perioperative chemotherapy treatment for gastroesophageal cancer. The gastroesophageal cancer may, for example, be an operable gastroesophageal cancer, or a gastroesophageal cancer for which local therapy is indicated. The gastroesophageal cancer may, for example, be adenocarcinoma of the thoracic esophagus of EGJ. The additional anti-cancer therapy may, for example, comprise fluorouracil, leucovorin, oxaliplatin, docetaxel, fluoropyrimidine and/or cisplatin. The additional anti-cancer therapy may, for example, comprise combination therapy with: fluorouracil, leucovorin, oxaliplatin, and docetaxel; fluoropyrimidine and oxaliplatin; or fluorouracil and cisplatin.

[0116] The additional anti-cancer therapy may, for example, comprise a drug (or a combination of drugs) previously described for use in definitive chemoradiation for gastroesophageal cancer. The additional anti-cancer therapy may comprise a drug, or a combination of drugs, previously described as a standard-of-care definitive chemoradiation treatment for gastroesophageal cancer. The gastroesophageal cancer may, for example, be an operable gastroesophageal cancer, or a gastroesophageal cancer for which local therapy is indicated. The additional anti-cancer therapy may, for example, comprise paclitaxel, carboplatin, fluorouracil, oxaliplatin, cisplatin, docetaxel, irinotecan, and/or fluoropyrimidine (fluorouracil or capecitabine). The additional anti-cancer therapy may, for example, comprise combination therapy with: paclitaxel and carboplatin; fluorouracil and oxaliplatin; fluorouracil and cisplatin; cisplatin with docetaxel or paclitaxel; irinotecan and cisplatin; or paclitaxel and fluoropyrimidine (fluorouracil or capecitabine).

[0117] The additional anti-cancer therapy may, for example, comprise a drug (or a combination of drugs) previously described for use in postoperative therapy for gastroesophageal cancer. The additional anti-cancer therapy may comprise a drug, or a combination of drugs, previously described as a standard-of-care postoperative therapy treatment for gastroesophageal cancer. The gastroesophageal cancer may, for example, be an operable gastroesophageal cancer, or a gastroesophageal cancer for which local therapy is indicated. The additional anti-cancer therapy may, for example, comprise nivolumab, capecitabine, oxaliplatin, or fluorouracil. The additional anti-cancer therapy may, for example, comprise monotherapy with nivolumab, for instance after postoperative therapy with resection. The additional anti-cancer therapy may, for example, comprise combination therapy with: capecitabine and oxaliplatin; or fluorouracil and oxaliplatin.

[0118] The additional anti-cancer therapy may, for example, comprise a drug (or a combination of drugs) previously described for use in postoperative chemoradiation for gastroesophageal cancer. The additional anti-cancer therapy may comprise a drug, or a combination of drugs, previously described as a standard-of-care postoperative chemoradiation treatment for gastroesophageal cancer. The gastroesophageal cancer may, for example, be an operable gastroesophageal cancer, or a gastroesophageal cancer for which local therapy is indicated. The additional anti-cancer therapy may, for example, comprise fluoropyrimidine (i.e. fluorouracil or capecitabine). The additional anti-cancer therapy may, for example, comprise monotherapy with fluorouracil or capecitabine, for instance before and/or after fluoropyrimidine-based chemoradiation.

[0119] The additional anti-cancer therapy may, for example, comprise a drug (or a combination of drugs) previously described for first-line therapy of gastroesophageal cancer. The additional anti-cancer therapy may comprise a drug, or a combination of drugs, previously described as a standard-of-care first-line treatment for gastroesophageal cancer. The gastroesophageal cancer may, for example, be unresectable locally advanced, recurrent, or metastatic gastroesophageal cancer. Local therapy for the unresectable locally advanced, recurrent, or metastatic gastroesophageal cancer may not be indicated. The additional anti-cancer therapy may, for example, comprise fluoropyrimidine (fluorouracil or capecitabine), oxaliplatin, trastuzumab, cisplatin, nivolumab, pembrolizumab, ipilimumab, irinotecan, paclitaxel, carboplatin and/or docetaxel. The additional anti-cancer therapy may, for example, comprise combination therapy with: fluoropyrimidine (fluorouracil or capecitabine) and oxaliplatin and trastuzumab (e.g. for HER2 overexpression positive adenocarcinoma); fluoropyrimidine (fluorouracil or capecitabine) and cisplatin and trastuzumab (e.g. for HER2 overexpression positive adenocarcinoma); fluoropyrimidine (fluorouracil or capecitabine), oxaliplatin, and nivolumab for adenocarcinoma (e.g. for HER2 overexpression negative adenocarcinoma); fluoropyrimidine (fluorouracil or capecitabine), oxaliplatin, and nivolumab (e.g. for HER2 overexpression negative squamous cell carcinoma); fluoropyrimidine (fluorouracil or capecitabine), cisplatin, and nivolumab (e.g. for HER2 overexpression negative squamous cell carcinoma); fluoropyrimidine (fluorouracil or capecitabine), oxaliplatin, and pembrolizumab (e.g. for HER2 overexpression negative cancers); fluoropyrimidine (fluorouracil or capecitabine), cisplatin, and pembrolizumab (e.g. for HER2 overexpression negative cancers); fluoropyrimidine (fluorouracil or capecitabine) and oxaliplatin (e.g. for HER2 overexpression negative cancers); fluoropyrimidine (fluorouracil or capecitabine) and cisplatin (e.g. for HER2 overexpression negative cancers); nivolumab and ipilimumab (e.g. for HER2 overexpression negative squamous cell carcinoma); fluoropyrimidine (fluorouracil or capecitabine) and cisplatin and trastuzumab and pembrolizumab (e.g. for HER2 overexpression positive adenocarcinoma); fluoropyrimidine (fluorouracil or capecitabine) and oxaliplatin and trastuzumab and pembrolizumab (e.g. for HER2 overexpression positive adenocarcinoma); fluorouracil and irinotecan; paclitaxel with or without cisplatin or carboplatin; docetaxel with or without cisplatin; fluoropyrimidine (fluorouracil or capecitabine); docetaxel, cisplatin or oxaliplatin, and fluorouracil; or docetaxel, carboplatin, and fluorouracil.

[0120] The additional anti-cancer therapy may, for example, comprise a drug (or a combination of drugs) previously described for second-line or subsequent-line therapy of gastroesophageal cancer. The additional anti-cancer therapy may comprise a drug, or a combination of drugs, previously described as a standard-of-care second-line or subsequent-line treatment for gastroesophageal cancer. The gastroesophageal cancer may, for example, be unresectable locally advanced, recurrent, or metastatic gastroesophageal cancer. Local therapy for the unresectable locally advanced, recurrent, or metastatic gastroesophageal cancer may not be indicated. The additional anti-cancer therapy may, for example, comprise dostarlimab-gxly, nivolumab, pembrolizumab, docetaxel, paclitaxel, irinotecan, entrectinib, larotrectinib, ramucirumab, fam-trastuzumab, deruxtecan-nxki, fluorouracil, and/or cisplatin. The additional anti-cancer therapy may, for example, comprise monotherapy with dostarlimab-gxly (e.g. for MSI-H or dMMR tumours), nivolumab (e.g. for esophageal squamous cell carcinoma), pembrolizumab (e.g. for MSI-H or dMMR tumours, or for TMB high (10 mutations/megabase) tumours, or for second-line therapy for esophageal squamous cell carcinoma with PD-L1 expression levels by CPS of 10), docetaxel, paclitaxel, irinotecan, entrectinib (e.g. for NTRK gene fusion-positive tumours) or larotrectinib (e.g. for NTRK gene fusion-positive tumours). The additional anti-cancer therapy may, for example, comprise combination therapy with: ramucirumab and paclitaxel (e.g. for adenocarcinoma such as EGJ adenocarcinoma or esophageal adenocarcinoma); fam-trastuzumab deruxtecan-nxki (e.g. for HER2 overexpression positive adenocarcinoma); fluorouracil and irinotecan; ramucirumab for adenocarcinoma (e.g. for EGJ adenocarcinoma or esophageal adenocarcinoma); irinotecan and cisplatin; fluorouracil and irinotecan and ramucirumab (e.g. for adenocarcinoma); irinotecan and ramucirumab (e.g. for adenocarcinoma); or docetaxel and irinotecan.

[0121] The additional anti-cancer therapy may, for example, comprise a drug (or a combination of drugs) previously described for third-line or subsequent-line therapy of gastroesophageal cancer. The additional anti-cancer therapy may comprise a drug, or a combination of drugs, previously described as a standard-of-care third-line or subsequent-line treatment for gastroesophageal cancer. The gastroesophageal cancer may, for example, be unresectable locally advanced, recurrent, or metastatic gastroesophageal cancer. Local therapy for the unresectable locally advanced, recurrent, or metastatic gastroesophageal cancer may not be indicated. The additional anti-cancer therapy may, for example, comprise trifluridine and/or tipiracil. The additional anti-cancer therapy may, for example, comprise combination therapy with trifluridine and tipiracil (e.g. for EGJ adenocarcinoma).

[0122] The population of modified T cells and the one or more additional anti-cancer therapies may be administered any number of times, and in any order. As set out above, the population of modified T cells may, for example, be administered as a single dose. An additional anti-cancer therapy may, for example, be administered (a) before the modified T cells, (b) at the same time as the modified T cells, and/or (c) after the modified T cells. For instance, an additional anti-cancer therapy may, for example, be administered: (a); (b); (c); (a) and (b); (a) and (c); (b) and (c); or (a), (b) and (c). When the additional anti-cancer therapy is administered at the same time as the population of modified T cells, the additional anti-cancer therapy and the population of modified T cells may be comprised in the same composition or in separate compositions. In preferred aspects of the invention, administration of the additional anti-cancer therapy begins before administration of the population of modified T cells, or after administration of the population of modified T cells.

[0123] Administration of an additional anti-cancer therapy at the same time as the modified T cells may refer to administration of the additional anti-cancer therapy and the modified T cells at substantially the same time. In other words, a dose of the additional anti-cancer therapy may be administered at about the same time as a dose of the population of modified T cells. For instance, a dose of the additional anti-cancer therapy may be administered within about 2 hours, about 4 hours, about 6 hours, about 8 hours, about 12 hours, or about 24 hours of a dose of the population of modified T cells.

[0124] Administration of an additional anti-cancer therapy before the modified T cells may refer to administration of the additional anti-cancer therapy at any time before the modified T cells. In other words, a dose of the additional anti-cancer therapy may be administered at any time before a dose of the population of modified T cells. For instance, a dose of the additional anti-cancer therapy may be administered about 24 hours or more, about 36 hours or more, about 48 hours or more, about 72 hours or more, about 4 days or more, about 5 days or more, about 6 days or more, about one week or more, about two weeks or more, about three weeks or more, or about four weeks or more, about five weeks or more, about six weeks or more, about 8 weeks or more, or about 12 weeks or more, before the population of modified T cells. The additional anti-cancer therapy may, for example, be administered before the modified T cells in order to initiate treatment while autologous modified T cells are produced.

[0125] Administration of the additional anti-cancer therapy after the modified T cells may refer to administration of the anti-cancer therapy at any time after the modified T cells. In other words, a dose of the additional anti-cancer therapy may be administered at any time after a dose of the population of modified T cells. For instance, a dose of the additional anti-cancer therapy may be administered about 24 hours or more, about 36 hours or more, about 48 hours or more, about 72 hours or more, about 4 days or more, about 5 days or more, about 6 days or more, about one week or more, about two weeks or more, about three weeks or more, or about four weeks or more, about five weeks or more, about six weeks or more, about 8 weeks or more, or about 12 weeks or more, after the population of modified T cells. The additional anti-cancer therapy may, for example, be administered after the modified T cells in order to initiate treatment while autologous modified T cells are produced.

[0126] If the additional anti-cancer therapy is administered before the population of modified T cells, the additional anti-cancer therapy may be continued after administration of the modified T cells. Thus, in one aspect of the disclosure, administration of the additional anti-cancer therapy begins before administration of the population of modified T cells, and continues after administration of the population of modified T cells. In other words, a dose of the additional anti-cancer therapy may be administered before a dose of the population of modified T cells, and one or more further doses of the additional anti-cancer therapy may be administered later. The doses of the additional anti-cancer therapy may, for example, be administered in accordance with a known treatment regime for the additional anti-cancer therapy. The purpose of the further doses of the additional anti-cancer therapy may be to maintain the effects achieved by administration of the initial dose. Each of the one or more further doses may comprise the same additional anti-cancer therapy as the initial dose, or a different additional anti-cancer therapy from the initial dose.

[0127] If the additional anti-cancer therapy is administered at the same time as the population of modified T cells, the additional anti-cancer therapy may be continued after administration of the modified T cells. Thus, in one aspect of the disclosure, administration of the additional anti-cancer therapy begins at the same time as administration of the population of modified T cells, and continues after administration of the population of modified T cells. In other words, a dose of the additional anti-cancer therapy may be administered at the same time as a dose of the population of modified T cells, and one or more further doses of the additional anti-cancer therapy may be administered later. The doses of the additional anti-cancer therapy may, for example, be administered in accordance with a known treatment regime for the additional anti-cancer therapy. The purpose of the further doses of the additional anti-cancer therapy may be to maintain the effects achieved by administration of the initial dose. Each of the one or more further doses may comprise the same additional anti-cancer therapy as the initial dose, or a different additional anti-cancer therapy from the initial dose.

[0128] If the additional anti-cancer therapy is administered after the population of modified T cells, the additional anti-cancer therapy may be continued after initial administration. Thus, in one aspect of the disclosure, administration of the additional anti-cancer therapy begins after administration of the population of modified T cells, and continues after initial administration of the additional anti-cancer therapy. In other words, a dose of the additional anti-cancer therapy may be administered after a dose of the population of modified T cells, and one or more further doses of the additional anti-cancer therapy may be administered later. The doses of the additional anti-cancer therapy may, for example, be administered in accordance with a known treatment regime for the additional anti-cancer therapy. The purpose of the further doses of the additional anti-cancer therapy may be to maintain the effects achieved by administration of the initial dose. Each of the one or more further doses may comprise the same additional anti-cancer therapy as the initial dose, or a different additional anti-cancer therapy from the initial dose.

[0129] In any case, the one or more further doses of the additional anti-cancer therapy may be administered at any appropriate interval. Any number of further doses of additional anti-cancer therapy may be administered, such as one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, 15 or more, 20 or more, 25 or more, 30 or more, 35 or more, 40 or more, 45 or more or 50 or more further doses. Further doses may be administered until disease progression, unacceptable toxicity, withdrawal of consent, or death. Suitable dosage intervals for additional anti-cancer therapy are known in the art and may be peculiar to the identity of the additional anti-cancer therapy.

[0130] For example, for paclitaxel, the one or more further doses may, for example, be administered on day 1, day 8 and day 15 of a 28 day cycle. The one or more further doses may be about once every one week (beginning one week from administration of the initial dose). Paclixtacel may be administered in a 4 week cycle, where it is administered about once every three weeks followed by one week without administration (3Q4W).

[0131] In a preferred aspect of the disclosure, the additional anti-cancer therapy comprises paclitaxel and/or ramucirumab. For example, the additional anti-cancer therapy may comprise paclitaxel, ramucirumab, or both paclitaxel and ramucirumab. Preferably, the additional anti-cancer therapy comprises paclitaxel and ramucirumab. The paclitaxel and/or ramucirumab (e.g. paclitaxel and ramucirumab) may, for example, be administered before the population of modified T cells. The paclitaxel and/or ramucirumab (e.g. paclitaxel and ramucirumab) may, for example, be administered after the population of modified T cells. As set out below, a checkpoint inhibitor (e.g. a PD-1 axis binding antagonist such as nivolumab or pembrolizumab) may additionally be administered to the individual. The checkpoint inhibitor (e.g. nivolumab or pembrolizumab) may, for example, be administered after the population of modified T cells. The checkpoint inhibitor (e.g. nivolumab or pembrolizumab) may, for example, be administered after the paclitaxel and/or ramucirumab (e.g. paclitaxel and ramucirumab). The checkpoint inhibitor (e.g. nivolumab) may, for example, be administered after (1) the population of modified T cells and (2) the paclitaxel and/or ramucirumab (e.g. paclitaxel and ramucirumab).

Checkpoint Inhibitors

[0132] The method may comprise administering a checkpoint inhibitor to the individual. Therefore, the method may comprise administering to the individual (i) a population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous T-cell receptor (TCR) capable of binding to a peptide antigen of MAGE-A4, and (ii) a checkpoint inhibitor. In other words, the method may comprise combination treatment with individual (i) a population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous T-cell receptor (TCR) capable of binding to a peptide antigen of MAGE-A4, and (ii) a checkpoint inhibitor. An additional anti-cancer therapy, such as a chemotherapy, may also be included in the combination as set out above.

[0133] The checkpoint inhibitor may be administered to the individual in the same line of therapy as the population of modified T cells. In As set out above, a line of therapy may refer to a particular treatment regimen for a cancer for which curative intent treatment has failed, or for which curative intent treatment may be inappropriate. Thus, the checkpoint inhibitor and the population of modified T cells may be administered as part of the same treatment regimen for a cancer for which curative intent treatment has failed, or for which curative intent treatment may be inappropriate. The checkpoint inhibitor and the population of modified T cells may be administered as part of a first-line treatment regimen. The checkpoint inhibitor and the population of modified T cells may be administered as part of a second-line treatment regimen. The second-line treatment regimen may, for example, be employed following failure of the first-line treatment regimen. The checkpoint inhibitor and the population of modified T cells may be administered as part of a third-line treatment regimen. The third-line treatment regimen may, for example, be employed following failure of the second-line treatment regimen.

[0134] Checkpoint inhibitor therapy is a form of cancer immunotherapy. The therapy targets immune checkpoints, key regulators of the immune system that when stimulated can dampen the immune response to an immunologic stimulus. Checkpoint therapy can block inhibitory checkpoints, restoring immune system function. Checkpoint inhibitors can target the molecules CTLA4, PD-1 and PD-L1.

[0135] The checkpoint inhibitor may, for example, target CTLA4. That is, the checkpoint inhibitor may comprise a CTLA4 blocker. CTLA4 blockers are known in the art and include, for example, ipilimumab.

[0136] Preferably, the checkpoint inhibitor comprises a PD-1 axis binding antagonist. PD-1 axis binding antagonists (e.g. nivolumab and pembrolizumab) are comprised in standard-of-care treatments for gastroesophageal cancer as discussed above.

[0137] Programmed cell death protein 1 (PD-1, also known as CD279) is a protein that is expressed on the surface of T cells and has a role in regulating immune responses by maintaining T cell homeostasis. Ligation of PD-1 to one of its ligands (PD-L1 or PD-L2) transmits an inhibitory signal within the T cell. In particular, PD-1-generated signals prevent phosphorylation of key TCR signalling intermediates, thereby terminating early TCR signalling and reducing T cell activation. T cell effector functions (such as proliferation, cytotoxicity and cytokine production) are reduced, and the ability to transition to memory T cells is impaired.

[0138] PD-L1 and PD-L2 are members of the B7 family. PD-L1 protein is upregulated on certain activated immune cells (such as macrophages, dendritic cells, T cells and B cells), and is also expressed upon certain normal tissues. PD-L1 is also highly expressed in many cancers. PD-L2 is expressed mainly by dendritic cells and some tumours. As many cancers express PD-1 ligands, the PD-1 axis has an established role in cancer immune evasion and tumour resistance.

[0139] Expression of PD-1 ligands by cancers, such as solid tumours, renders the tumour microenvironment immunosuppressive. The function of modified T cells infiltrating the tumour may therefore be inhibited. Endogenous anti-tumour T cell responses may also be inhibited. In this way, tumours are more able to evade the immune system. In the present disclosure, a PD-1 axis binding antagonist may be administered to counteract suppressive effects of PD-L1 and/or PD-L2 expression in the tumour microenvironment. By counteracting suppression, the function of modified and/or endogenous T cells may be sustained. That is, administration of a PD-1 axis binding antagonist may sustain the function of modified T cells comprised in the administration, and/or their descendants. Administration of a PD-1 axis binding antagonist may sustain the function of endogenous T cells in the individual. Administration of a PD-1 axis binding antagonist may sustain the function of modified T cells comprised in the administration (and/or their descendants), and of endogenous T cells in the individual. In any case, the endogenous T cells may, for example, be comprised in the tumour microenvironment. For instance, the endogenous T cells may be tumour infiltrating lymphocytes (TILs).

[0140] Sustaining the function of T cells may refer, for example, to maintaining, restoring and/or enhancing T cell function. Sustaining T cell function may, for example, refer to sustaining T cell activation. In this way, the duration of an effective T cell response may be extended. In other words, sustained activation maybe associated with an improved duration of effector function (such as cytokine production, cytotoxicity and/or proliferation). Sustained activation may also assist the T cells' ability to transition to memory T cells. The generation of memory T cells is advantageous, as it permits anti-tumour immunity to be maintained in the long-term e.g. for months or years. Methods for determining activation, cytokine production, cytotoxicity, proliferation, and generation of memory T cells are well-known in the art.

[0141] Administration of the PD-1 axis binding antagonist may sustain the function of the modified T cells and/or endogenous T cells by reducing exhaustion. Exhaustion may be reduced within the population of modified T cells, and/or within T cells descended from the population of modified T cells. Exhaustion may be reduced within endogenous T cells in the individual. Exhaustion may be reduced (i) within the population of modified T cells and/or within T cells descended from the population of modified T cells, and (ii) within endogenous T cells in the individual. Exhausted T cells typically express high levels of PD-1, and experience a loss of function. For instance, exhausted T cells may have reduced ability to produce cytokines such as IL-2 or TNF. Exhausted T cells may have reduced proliferative capacity. Exhausted T cells may have reduced cytotoxic potential. Ultimately, exhausted T cells may be targeted for destruction. Exhaustion therefore causes loss of T cell function, or loss of T cells themselves, which is disadvantageous to tumour immunity. Reducing T cell exhaustion may therefore improve therapeutic outcome.

[0142] In the context of the disclosure, a PD-1 axis binding antagonist is a molecule that inhibits the interaction of PD-1 with a PD-1 ligand, and/or transduction of a signal resulting from the interaction of PD-1 with a PD-1 ligand. The PD-1 ligand may be PD-L1 or PD-L2. The PD-1 axis binding antagonist may, for example, reduce or prevent the interaction of PD-1 with a PD-1 ligand. The PD-1 axis binding antagonist may, for example, reduce or prevent transduction of a signal resulting from the interaction of PD-1 with a PD-1 ligand. The PD-1 axis binding antagonist may block, inhibit or reduce the biological activity of PD-1 and/or a PD-1 ligand.

[0143] By inhibiting the interaction of PD-1 with a PD-1 ligand, and/or transduction of a signal resulting from the interaction of PD-1 with a PD-1 ligand, the PD-1 axis binding antagonist may sustain (e.g. maintain, restore or enhance) the function of endogenous T cells. In the same way, the PD-1 axis binding antagonist may sustain (e.g. maintain, restore or enhance) the function of modified T cells administered to the individual. Sustained function may, for example, be indicated by maintenance of, or improvements in, T-cell proliferation, cytokine production, target cell killing, activation, CD28 signalling, ability to infiltrate tumour, ability to recognise and bind to dendritic cell presented antigen, and/or ability to produce interferon. In this way, the PD-1 axis binding antagonist counteracts the immunosuppressive nature of the tumour microenvironment.

[0144] The PD-1 axis binding antagonist may, for example, be a PD-1 binding antagonist. That is, the PD-1 axis binding antagonist may inhibit (e.g. prevent or reduce) binding of PD-1 to its binding partners. For example, the PD-1 axis binding antagonist may inhibit the binding of PD-1 to PD-L1, PD-L2, or both PD-L1 and PD-L2. The PD-1 binding antagonist may, for example, be an antibody that binds to PD-1, or an antigen-binding variant or fragment thereof. The PD-1 binding antagonist may, for example, be an antibody that binds to SEQ ID NO: 14, or an antigen-binding variant or fragment thereof. Antibodies that bind to PD-1 are well-known in the art and include, for example, nivolumab, pembrolizumab, cemiplimab, dostarlimab, JTX-4014, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, retifanlimab (INCMGA00012), AMP-224, MEDI0680 (AMP-514), sasanlimab, budigalimab, ezabenlimab (BI 754091), and zimberelimab (AB122). The PD-1 axis binding antagonist may therefore be nivolumab, pembrolizumab, cemiplimab, dostarlimab, JTX-4014, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, retifanlimab (INCMGA00012), AMP-224, MEDI0680 (AMP-514), sasanlimab, budigalimab, ezabenlimab (BI 754091) or zimberelimab (AB122), or any combination thereof. The PD-1 axis binding antagonist may, for example, be nivolumab. The PD-1 axis binding antagonist may, for example, be pembrolizumab. The PD-1 binding antagonist may alternatively be any immunoadhesin, protein or oligopeptide that inhibits, decreases, prevent, or interferes with signal transduction due to the interaction of PD-1 with PD-L1 and/or PD-L2.

[0145] The heavy chain sequence and light chain sequence of nivolumab are set out in SEQ ID NOs: 17 and 18 respectively. The heavy chain sequence and light chain sequence of pembrolizumab are set out in SEQ ID NOs: 19 and 20 respectively. The heavy chain sequence and light chain sequence of cemiplimab are set out in SEQ ID NOs: 21 and 22 respectively. The heavy chain sequence and light chain sequence of dostarlimab are set out in SEQ ID NOs: 31 and 32 respectively. A skilled person equipped with the heavy and light chain sequences of a given antibody may identify antigen-binding variants or fragments of the antibody using methods routine in the art. An antigen-binding variant or fragment of nivolumab may, for example, comprise the three CDRs comprised in SEQ ID NO: 17 and the three CDRs comprised in SEQ ID NO: 18. An antigen-binding variant or fragment of pembrolizumab may, for example, comprise the three CDRs comprised in SEQ ID NO: 19 and the three CDRs comprised in SEQ ID NO: 20. An antigen-binding variant or fragment of cemiplimab may, for example, comprise the three CDRs comprised in SEQ ID NO: 21 and the three CDRs comprised in SEQ ID NO: 22. An antigen-binding variant or fragment of dostarlimab may, for example, comprise the three CDRs comprised in SEQ ID NO: 31 and the three CDRs comprised in SEQ ID NO: 32. Methods for identifying CDRs within a heavy chain or light chain sequence are routine in the art.

[0146] The PD-1 axis binding antagonist may, for example, be a PD-L1 binding antagonist. That is, the PD-1 axis binding antagonist may inhibit (e.g. prevent or reduce) binding of PD-L1 to a binding partner. For example, the PD-1 axis binding antagonist may inhibit the binding of PD-L1 to PD-1. The PD-L1 binding antagonist may, for example, be an antibody that binds PD-L1, or an antigen-binding variant or fragment thereof. The PD-L1 binding antagonist may, for example, be an antibody that binds to SEQ ID NO: 15, or an antigen-binding variant or fragment thereof. Antibodies that bind to PD-L1 are well-known in the art and include, for example, durvalumab, atezolizumab, avelumab, BMS 936559 (MDX-1105), envafolimab (KN035) and cosibelimab (CK-301). The PD-L1 axis binding antagonist may therefore be durvalumab, atezolizumab, avelumab, BMS 936559 (MDX-1105), envafolimab (KN035) or cosibelimab (CK-301), or any combination thereof. The PD-L1 binding antagonist may alternatively be any immunoadhesin, protein or oligopeptide that inhibits, decreases, prevent, or interferes with signal transduction due to the interaction of PD-L1 with PD-1.

[0147] The heavy chain sequence and light chain sequence of durvalumab are set out in SEQ ID NOs: 23 and 24 respectively. The heavy chain sequence and light chain sequence of atezolizumab are set out in SEQ ID NOs: 25 and 26 respectively. The heavy chain sequence and light chain sequence of avelumab are set out in SEQ ID NOs: 27 and 28 respectively. The heavy chain sequence and light chain sequence of BMS 936559 (MDX-1105) are set out in SEQ ID NOs: 29 and 30 respectively. A skilled person equipped with the heavy and light chain sequences of a given antibody may identify antigen-binding variants or fragments of the antibody using methods routine in the art. An antigen-binding variant or fragment of durvalumab may, for example, comprise the three CDRs comprised in SEQ ID NO: 23 and the three CDRs comprised in SEQ ID NO: 24. An antigen-binding variant or fragment of atezolizumab may, for example, comprise the three CDRs comprised in SEQ ID NO: 25 and the three CDRs comprised in SEQ ID NO: 26. An antigen-binding variant or fragment of avelumab may, for example, comprise the three CDRs comprised in SEQ ID NO: 27 and the three CDRs comprised in SEQ ID NO: 28. An antigen-binding variant or fragment of BMS 936559 (MDX-1105) may, for example, comprise the three CDRs comprised in SEQ ID NO: 29 and the three CDRs comprised in SEQ ID NO: 30. Methods for identifying CDRs within a heavy chain or light chain sequence are routine in the art.

[0148] The PD-1 axis binding antagonist may, for example, be a PD-L2 binding antagonist. That is, the PD-1 axis binding antagonist may inhibit (e.g. prevent or reduce) binding of PD-L2 to a binding partner. The PD-L2 binding antagonist may, for example, be an antibody that binds PD-L2, or an antigen-binding variant or fragment thereof. The PD-L2 binding antagonist may, for example, be an antibody that binds to SEQ ID NO: 16, or an antigen-binding variant or fragment thereof. The PD-L2 binding antagonist may alternatively be any immunoadhesin, protein or oligopeptide that inhibits, decreases, prevent, or interferes with signal transduction due to the interaction of PD-L2 with PD-1.

[0149] When the PD-1 axis binding antagonist is an antibody (such as a known antibody, or an antigen-binding variant thereof), the PD-1 axis binding antagonist may be a monoclonal antibody, a human or humanised antibody, a full-length antibody, a diabody, a linear antibody, or a single-chain antibody molecule, for example. The antibody isotype may be selected from any of the five classes of immunoglobulins: IgA, IgD, IgE, IgG and IgM, having heavy chains designated alpha, delta, epsilon, gamma and mu (M), respectively. The gamma and alpha class antibodies may be of any of subclasses: IgG1, IgG2A, IgG2B, IgG3, IgG4, IgAQ1 and IgA2. When the PD-1 axis binding antagonist is an antigen-binding fragment of an antibody, the PD-1 axis binding antagonist may be a Fv, Fab, Fab, Fab-SH, F(ab) 2, or scFv, for example.

[0150] When the PD-1 axis binding antagonist is an immunoadhesin, the immunoadhesin may comprise an adhesin domain conferring binding activity for a PD-1 axis component (e.g. PD-1, PD-L1, or PD-L2) and an immunoglobulin constant domain. The immunoglobulin constant domain may be from any isotype, such as IgG1, IgG2, IgG2A, IgG2B, IgG3, IgG4 subtypes, IgA, IgA1, IgA2, IgE, IgD or IgM. The immunoglobulin constant domain may, for example, comprise (i) the hinge, CH2 and CH3, or (ii) the hinge, CH1, CH2 and CH3 regions of an immunoglobulin molecule. Accordingly, the immunoadhesin may comprise (a) the extracellular or PD-1 binding portions of PD-L1 or PD-L2, or the extracellular or PD-L1 or PD-L2 binding portions of PD-1, fused to (b) a constant domain of an immunoglobulin sequence.

[0151] At least 1% of the gastroesophageal cancer cells from the individual may express PD-L1, such as at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70% or at least 80%. The gastroesophageal cancer from the individual may express PD-L1 with a tumour proportion score (TPS) of greater than or equal to () 1%, such as 2%, 10% or as 50%. The percentage of cells that express PD-L1 may be determined by any means known to the skilled person, such as IHC, flow-cytometry or ELISA.

[0152] At least 1% of the gastroesophageal cancer cells from the individual may express PD-L2, such as at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70% or at least 80%. The gastroesophageal cancer from the individual may express PD-L2 with a tumour proportion score (TPS) of greater than or equal to () 1%, such as 2%, 10% or as 50%. The percentage of cells that express PD-L2 may be determined by any means known to the skilled person, such as IHC, flow-cytometry or ELISA.

[0153] The expression of PD-1, PD-L1 and/or PD-L2 in the gastroesophageal cancer may have an intensity of greater than or equal to () 1+, such as 2+ or 3+. The intensity score may be assessed by IHC staining of the tumour, with the scoring as follows: negative=no staining or staining in less than or equal to 10% of the cells stained; 1+=incomplete staining in 10% of cells stained; 2+=weak to moderate staining in 10% of cells stained; strong and complete staining in 10% of cells stained.

[0154] The population of modified T cells and the checkpoint inhibitor may be administered any number of times, and in any order. The population of modified T cells may, for example, be administered as a single dose. A checkpoint inhibitor may, for example, be administered (a) before the modified T cells, (b) at the same time as the modified T cells, and/or (c) after the modified T cells. For instance, checkpoint inhibitor may, for example, be administered: (a); (b); (c); (a) and (b); (a) and (c); (b) and (c); or (a), (b) and (c). When the checkpoint inhibitor is administered at the same time as the population of modified T cells, the checkpoint inhibitor and the population of modified T cells may be comprised in the same composition or in separate compositions.

[0155] Administration of a checkpoint inhibitor at the same time as the modified T cells may refer to administration of the checkpoint inhibitor and the modified T cells at substantially the same time. In other words, a dose of the checkpoint inhibitor may be administered at about the same time as a dose of the population of modified T cells. For instance, a dose of the checkpoint inhibitor may be administered within about 2 hours, about 4 hours, about 6 hours, about 8 hours, about 12 hours, or about 24 hours as a dose of the population of modified T cells.

[0156] Administration of a checkpoint inhibitor before the modified T cells may refer to administration of the checkpoint inhibitor at any time before the modified T cells. In other words, a dose of the checkpoint inhibitor may be administered at any time before a dose of the population of modified T cells. For instance, a dose of the checkpoint inhibitor may be administered about 24 hours or more, about 36 hours or more, about 48 hours or more, about 72 hours or more, about 4 days or more, about 5 days or more, about 6 days or more, about one week or more, about two weeks or more, about three weeks or more, or about four weeks or more, about five weeks or more, about six weeks or more, about 8 weeks or more, or about 12 weeks or more, before the population of modified T cells. The checkpoint inhibitor may, for example, be administered before the modified T cells in order to initiate treatment while autologous modified T cells are produced.

[0157] Administration of a checkpoint inhibitor after the modified T cells may refer to administration of the checkpoint inhibitor at any time after the modified T cells. In other words, a dose of the checkpoint inhibitor may be administered at any time after a dose of the population of modified T cells. For instance, a dose of the checkpoint inhibitor may be administered about 24 hours or more, about 36 hours or more, about 48 hours or more, about 72 hours or more, about 4 days or more, about 5 days or more, about 6 days or more, about one week or more, about two weeks or more, about three weeks or more, or about four weeks or more, about five weeks or more, about six weeks or more, about 8 weeks or more, or about 12 weeks or more, after the population of modified T cells.

[0158] If the checkpoint inhibitor is administered at the same time as the population of modified T cells, the checkpoint inhibitor may be continued after administration of the modified T cells. Thus, in one aspect of the disclosure, administration of the checkpoint inhibitor begins at the same time as administration of the population of modified T cells, and continues after administration of the population of modified T cells. In other words, a dose of the checkpoint inhibitor may be administered at the same time as a dose of the population of modified T cells, and one or more further doses of the checkpoint inhibitor may be administered later. The doses of the checkpoint inhibitor may, for example, be administered in accordance with a known treatment regime for the checkpoint inhibitor. The purpose of the further doses of the checkpoint inhibitor may be to maintain the effects achieved by administration of the initial dose. Each of the one or more further doses may comprise the same checkpoint inhibitor as the initial dose, or a different checkpoint inhibitor from the initial dose.

[0159] If the checkpoint inhibitor is administered after the population of modified T cells, the checkpoint inhibitor may be continued after initial administration. Thus, in one aspect of the disclosure, administration of the checkpoint inhibitor begins after administration of the population of modified T cells, and continues after initial administration of the checkpoint inhibitor. In other words, a dose of the checkpoint inhibitor may be administered after a dose of the population of modified T cells, and one or more further doses of the checkpoint inhibitor may be administered later. The doses of the checkpoint inhibitor may, for example, be administered in accordance with a known treatment regime for the checkpoint inhibitor. The purpose of the further doses of the checkpoint inhibitor may be to maintain the effects achieved by administration of the initial dose. Each of the one or more further doses may comprise the same checkpoint inhibitor as the initial dose, or a different checkpoint inhibitor from the initial dose.

[0160] In any case, the one or more further doses of the checkpoint inhibitor may be administered at any appropriate interval. Suitable dosage intervals for checkpoint inhibitors are known in the art and may be peculiar to the identity of the checkpoint inhibitor. For example, for a PD-1 axis binding antagonist such as nivolumab or pembrolizumab, the one or more further doses may, for example, be administered about once every two weeks (Q2W) beginning two weeks from administration of the initial dose. The one or more further doses may, for example, be administered about once every three weeks (Q3W) beginning three weeks from administration of the initial dose. The one or more further doses may, for example, be administered about once every four weeks (Q4W) beginning four weeks from administration of the initial dose. The one or more further doses may, for example, be administered about once every five weeks (Q5W) beginning five weeks from administration of the initial dose. The one or more further doses may, for example, be administered about once every six weeks (Q6W) beginning six weeks from administration of the initial dose. In a preferred aspect of the disclosure, the checkpoint inhibitor is nivolumab and one or more further doses is administered about once every four weeks (Q4W) beginning four weeks from administration of the initial dose.

[0161] Any number of further doses of checkpoint inhibitor may be administered, such as one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, 15 or more, 20 or more, 25 or more, 30 or more, 35 or more, 40 or more, 45 or more or 50 or more further doses. Further doses may be administered until disease progression, unacceptable toxicity, withdrawal of consent, or death.

Combination Treatment Protocols

[0162] Examples of combination treatment protocols are shown in FIGS. 1 to 5.

[0163] FIG. 1 concerns the treatment of gastroesophageal cancer that has relapsed following a first-line treatment, for instance an approved or standard-of-care first-line treatment. In this case, the individual may be administered (i) the population of modified T cells, (ii) an additional anti-cancer therapy, and optionally (iii) a PD-1 axis binding antagonist. Administration of the population of modified T cells may be preceded by lymphodepletion.

[0164] The additional anti-cancer therapy may, for example, be an approved or standard-of-care second-line treatment for gastroesophageal cancer. The standard-of-care second-line treatment may, for instance, comprise paclitaxel and/or ramucirumab. For example, the standard-of-care second-line treatment may comprise paclitaxel and/or ramucirumab. Treatment regimens for paclitaxel and/or ramucirumab are known in the art.

[0165] The population of modified T cells and the additional anti-cancer therapy may be administered in either order. For instance, in accordance with Cohort 2A (left of FIG. 1), the additional anti-cancer therapy may be administered before the population of modified T cells. Alternatively, in accordance with Cohort 2B (right of FIG. 1), the additional anti-cancer therapy may be administered after the population of modified T cells. In either case, administration of the additional anti-cancer therapy maybe continued after administration of the modified T cells. In other words, one or more doses of the additional anti-cancer therapy may be administered after administration of the modified T cells.

[0166] If a PD-1 axis binding antagonist is administered, it is typically administered after the population of modified T cells and the additional anti-cancer therapy. For instance, in accordance with Cohort 2A (left of FIG. 1), the individual may be administered with the additional anti-cancer therapy, then the population of modified T cells, then a PD-1 axis binding antagonist. However, it is also possible that the individual may be administered with the population of modified T cells, then the additional anti-cancer therapy, then a PD-1 axis binding antagonist. The PD-1 axis binding antagonist may, for example, be nivolumab or pembrolizumab.

[0167] In FIG. 1, administration of the modified T cells and the additional anti-cancer therapy (with or without a PD-1 axis binding antagonist) may represent second-line treatment of the gastroesophageal cancer.

[0168] FIG. 2 concerns the treatment of gastroesophageal cancer that has relapsed following a second-line treatment, for instance an approved or standard-of-care second-line treatment. Implicitly, the gastroesophageal cancer has also relapsed following a first-line treatment, for instance an approved standard-of-care first-line treatment. To treat gastroesophageal cancer that has relapsed following a second-line treatment, the individual may be administered (i) the population of modified T cells and (ii) a checkpoint inhibitor. Administration of the population of modified T cells may be preceded by lymphodepletion.

[0169] The checkpoint inhibitor may, for example, comprise a PD-1 axis binding antagonist. The PD-1 axis binding antagonist may, for example, be nivolumab or pembrolizumab. Treatment regimens for PD-1 axis binding antagonists such as nivolumab and pembrolizumab are known in the art.

[0170] The checkpoint inhibitor may, for example, be administered before the population of modified T cells, after the population of modified T cells., or at the same time (or substantially the same time) as the modified T cells. In one aspect of the disclosure, a dose of the checkpoint inhibitor may be administered at about the same time as a dose of the population of modified T cells. For instance, a dose of the checkpoint inhibitor may be administered within about 2 hours, about 4 hours, about 6 hours, about 8 hours, about 12 hours, or about 24 hours of a dose of the population of modified T cells. Administration of the checkpoint inhibitor may, for example, be continued after administration of the modified T cells. In one aspect of the disclosure, a dose of the checkpoint inhibitor may be administered at about the same time as a dose of the population of modified T cells, and one or more further doses of the checkpoint inhibitor may be administered later. The doses of the checkpoint inhibitor may, for example, be administered in accordance with a known treatment regime for the checkpoint inhibitor. Treatment regimens for PD-1 axis binding inhibitors (e.g. nivolumab or pembrolizumab) are known in the art.

[0171] An additional anti-cancer therapy may optionally be administered in the protocol shown in FIG. 2. Additional anti-cancer therapies are described above.

[0172] In FIG. 2, administration of the modified T cells and the checkpoint inhibitor may represent third-line or subsequent-line treatment of the gastroesophageal cancer.

[0173] FIGS. 4 and 5 concerns the treatment of gastroesophageal cancer that (A) has relapse following curative intent treatment of locally advanced cancer, or (B) is the first diagnosis of unresectable locally advanced cancer or metastatic cancer. In this case, the individual may be administered (i) an oxaliplatin-based therapy and (ii) the population of modified T cells. Optionally, the individual may be administered, (iii) a PD-1 axis binding antagonist and/or an additional anti-cancer therapy. Administration of the population of modified T cells may be preceded by lymphodepletion.

[0174] The PD-1 axis binding antagonist may, for example, be an approved or standard-of-care treatment for gastroesophageal cancer. The PD-1 axis binding antagonist may, for example, be pembrolizumab or nivolumab. The additional anti-cancer therapy may, for example, be an approved or standard-of-care treatment for gastroesophageal cancer. The additional anti-cancer therapy may, for example, comprise fluorouracil (5FU).

[0175] The oxaliplatin-based therapy may be administered before the population of modified T cells. Administration of the additional anti-cancer therapy maybe continued after administration of the modified T cells. In other words, one or more doses of the additional anti-cancer therapy may be administered after administration of the modified T cells.

[0176] If a PD-1 axis binding antagonist and/or an additional anti-cancer therapy is administered, it is typically administered after the oxaliplatin-based therapy and the population of modified T cells. For instance, the individual may be administered with oxaliplatin-based therapy, then the modified T cells, and then a PD-1 axis binding antagonist and/or an additional anti-cancer therapy.

[0177] In FIGS. 4 and 5, administration of the modified T cells in combination with an oxaliplatin based therapy (and optionally an additional anti-cancer therapy and/or a PD-1 axis binding antagonist) may represent first-line treatment of the gastroesophageal cancer.

[0178] In combination therapies, administration of the modified T cells may be advantageous. In particular, administration of modified T cells may allow the dose of a checkpoint inhibitor or an additional anti-cancer therapy to be reduced. In turn, cytokine release syndrome (CRS) and/or cytopenia may be reduced, especially when the additional anti-cancer therapy is a chemotherapy.

[0179] In combination therapies, administration of a checkpoint inhibitor or an additional anti-cancer therapy may be advantageous. In particular, the checkpoint inhibitor or additional anti-cancer therapy may be implemented while an autologous population of modified T cells is produced, thereby allowing treatment to begin as soon as possible. This may improve therapeutic outcomes. Administration of the checkpoint inhibitor or additional anti-cancer therapy and the modified T cells may also have an adjunctive effect.

Administration

[0180] The population of modified T cells may, for example, be administered to the individual as soon as possible after diagnosis of relapse gastroesophageal cancer, such as relapsed gastroesophageal cancer. The population may be administered to the individual, for example, less than about 150 days after diagnosis of the gastroesophageal cancer, such as less than about 125 days, less than about 100 days, less than about 90 days, less than about 80 days or less than about 70 days after diagnosis of the gastroesophageal cancer. The population may be administered to the individual, for example, about 30 to about 150 days after diagnosis of the gastroesophageal cancer, such as about 40 to about 125, about 50 to about 100, about 90, about 85, about 80, about 75, about 70, about 65 or about 60 days after diagnosis of the gastroesophageal cancer.

[0181] Typically, the population of modified T cells is administered as a single dose. One or more (such as two or more, three or more, four or more, or five or more) further doses may though be administered depending on patient factors and the judgement of the practitioner.

[0182] The population may comprise any number of modified T cells that will be therapeutically effective. The number of modified T cells for a given individual may depend on factors such as the cancer to be treated, the severity or stage of the cancer, the age of the patient and so on. The number to be administered may thus depend on the judgement of the practitioner and may be peculiar to each subject. By way of example, the population may comprise about 0.810.sup.9 to about 1010.sup.9 modified T cells, such as about 0.810.sup.9 to about 1.210.sup.9 modified T cells, about 1.210.sup.9 to about 610.sup.9 modified T cells, or about 1.010.sup.9 to 1010.sup.9 modified T cells. The population may, for example, comprise 1.010.sup.9 modified T cells, about 5.010.sup.9 modified T cells, or about 1010.sup.9 modified T cells.

[0183] Typically, the population of modified T cells is administered intravenously. Any suitable route may though be used, for instance intramuscular, subcutaneous, intradermal, transdermal, or intraperitoneal routes.

[0184] The population of modified T cells may be administered in combination with a checkpoint inhibitor and/or an anti-cancer therapy, as set out above. The checkpoint inhibitor or additional anti-cancer therapy may be administered by any route suitable for the given therapy, such as intravenous, intramuscular, subcutaneous, intradermal, transdermal, or intraperitoneal. As set out above, one or more doses of the checkpoint inhibitor or anti-cancer therapy may be administered.

[0185] Each dose of the checkpoint inhibitor or anti-cancer therapy may comprise any therapeutically effective amount of the checkpoint inhibitor or anti-cancer therapy. The amount for a given individual may depend on factors such as the cancer to be treated, the severity or stage of the cancer, the age of the patient and so on. The amount to be administered may thus depend on the judgement of the practitioner and may be peculiar to each subject.

[0186] By way of example, checkpoint inhibitor may be a PD-1 axis binding antagonist, such as nivolumab. The initial dose of the PD-1 axis binding antagonist, and/or any further dose of the PD-1 axis binding antagonist, may comprise about 200 mg to about 700 mg of nivolumab, such as about 200 mg to about 500 mg of nivolumab. For instance, the initial dose of nivolumab, and/or any further dose of nivolumab, may comprise about 240 mg, about 360 mg or about 480 mg of nivolumab. The initial dose of nivolumab, and/or any further dose of nivolumab, may, for example, comprise about 200 mg to about 500 mg of the nivolumab, such as about 480 mg of the nivolumab. In a preferred aspect of the disclosure, the initial dose of nivolumab comprises 240 mg, 360 mg or 480 mg nivolumab, and any further dose of the PD-1 axis binding antagonist, comprises 240 mg Q2W of nivolumab or 480 mg Q4W of nivolumab. Nivolumab may be administered at 3 mg/kg Q2W.

[0187] In some aspects of the disclosure, the method comprises administering lymphodepleting chemotherapy to the individual prior to administration of the population of modified T cells. That is, lymphodepleting chemotherapy may be administered before step (a). Lymphodepleting chemotherapy may, for example, be administered from about 14 days before step (a) to about 1 day before step (a), such as about 13 days before step (a) to about 2 days before step (a), about 12 days before step (a) to about 3 days before step (a), about 11 days before step (a) to about 4 days before step (a), about 10 days before step (a) to about 5 days before step (a), about 9 days before step (a) to about 6 days before step (a), about 8 days before step (a) to about 7 days before step (a), about 10 days before step (a) to about 1 day before step (a), about 9 days before step (a) to about 2 days before step (a), about 8 days before step (a) to about 3 days before step (a), about 7 days before step (a) to about 4 days before step (a), or about 6 days before step (a) to about 5 days before step (a). Preferably, lymphodepleting chemotherapy is administered from about 7 days before step (a) to about 4 days before step (a). The purpose of lymphodepleting chemotherapy may be to deplete the lymphocyte compartment of the individual, so as to provide space into which the adoptively-transferred modified T cells can expand. In this way, the effects of a given dose of the modified T cells can be maximised. The lymphodepleting chemotherapy may comprise any suitable lymphotoxic agent. Lymphotoxic agents and suitable dosages are known in the art. The lymphodepleting chemotherapy may, for example, comprise fludarabine and/or cyclophosphamide. Typically, the lymphodepleting chemotherapy is administered intravenously. Any suitable route may though be used, for instance intramuscular, subcutaneous, intradermal, transdermal, or intraperitoneal routes.

Medicaments and Medical Uses

[0188] The disclosure provides a population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous T-cell receptor capable of binding to a peptide antigen of MAGE-A4 for use in a method of treating gastroesophageal cancer in an individual. Any of the aspects described above in connection with the method of the disclosure may also apply to the population for use.

[0189] The disclosure also provides the use of a population of modified T cells in the manufacture of a medicament for use in a method of treating gastroesophageal cancer in an individual, wherein the modified T cells comprise a heterologous CD8 co-receptor and a heterologous T-cell receptor capable of binding to a peptide antigen of MAGE-A4. Any of the aspects described above in connection with the method of the disclosure may also apply to this use of the population.

[0190] The disclosure also provides the use of a population of modified T cells in a method of treating gastroesophageal cancer in an individual, wherein the modified T cells comprise a heterologous CD8 co-receptor and a heterologous T-cell receptor capable of binding to a peptide antigen of MAGE-A4. Any of the aspects described above in connection with the method of the disclosure may also apply to this use of the population.

EXAMPLES

Introduction

[0191] ADP-A2M4CD8 specific peptide enhanced affinity receptor (SPEAR) T cells are genetically engineered to target the tumour antigen MAGE-A4 in the context of the appropriate human leukocyte antigen (HLA) expression. ADP-A2M4CD8 are autologous CD4 and CD8 positive T cells that have been transduced with a self-inactivating (SIN) lentiviral vector expressing a high affinity MAGE-A4 specific T cell receptor (TCR) and an additional CD8a co-receptor.

[0192] The affinity-optimised TCR (ADP-A2M4 TCR) comprises an alpha chain variable domain comprised in SEQ ID NO: 2, and a beta chain variable domain comprised in SEQ ID NO: 3. When expressed in a T cell, the signal peptides are cleaved from SEQ ID NOs: 2 and 3 prior to surface expression. A2M4 TCR targets the tumour antigen MAGE-A4 and activates engineered T cells. It recognizes the MAGE-A4230-239 (GVYDGREHTV; SEQ ID NO: 1) peptide sequence derived from MAGE-A4, when presented in the HLA-A*02-GVYDGREHTV antigen complex.

[0193] The CD8 co-receptor comprised in ADP-A2M4CD8 SPEAR T cells is designed to provide additional functionality to CD4 T cells. Because CD4+ T cells have a weak effector function in response to Class I antigens, a CD8 co-receptor was introduced alongside the TCR, in order to increase TCR binding avidity and enhance the polyfunctional response of engineered CD4+ T cells against MAGE-A4 positive tumour. The co-expression of CD8 adds CD8+ killer T cell capability to CD4+ helper T cells, while also maintaining/enhancing the helper cell capabilities of CD4+ T cells. The addition of a CD8 co-receptor directly impacts TCR binding to the HLA-peptide complex in CD4+ T cells, enhancing CD4+ T cell effector function. ADP-A2M4CD8 SPEAR T cells are therefore designed to improve upon ADP-A2M4 expressing T cells.

[0194] This has been confirmed in preclinical in vitro assays, in which ADP-A2M4CD8 showed a clear improvement in T cell activation (when cultured with antigen positive cells) relative to ADP-A2M4 expressing T cells, as measured by increased CD40L surface expression, particularly in the CD4+ fraction. When dendritic cells (DCs) were included in co-cultures, a marked improvement was seen with ADP-A2M4CD8 T cells. Cytokine release from both DCs (IL-12, MIG) and T cells (IFNy, IL-2 and other Th1) was improved compared to cultures containing the ADP-A2M4 cells. Additionally, a conversion of CD4+ T cells was seen, from being unable to kill MAGE-A4 positive 3D microspheres, to having an effective cytotoxic function when transduced with ADP-A2M4CD8. Therefore, CD4+ T cells transduced with ADP-A2M4CD8 display not only CD4+ helper functions, but also improved T cell effector functions.

[0195] While 77% of individuals having gastroesophageal cancer are alive five years after diagnosis, treatment options are suboptimal for individuals with advanced/metastatic disease. Less than 25% of individuals treated with chemotherapy make it to third-line treatment, and the third-line median progression-free survival is around 4 to 7 months.

[0196] An improved regimen for treatment of gastroesophageal cancer is desired. Preliminary clinical trials results in connection with ADP-A2M4CD8 in 13 individuals with gastroesophageal demonstrate confirmed responses in two individuals, and clear antitumour activity with an encouraging disease control rate (10 out of 13 individuals).

[0197] Examples 1 and 2 consider regimens in which ADP-A2M4CD8 is used as a second-line treatment or third-line treatment for gastroesophageal cancer respectively.

Example 1. Second-Line Treatment of Gastroesophageal Cancer

[0198] Subjects are selected for treatment with ADP-A2M4CD8. In brief, subjects eligible for selection must have been previously treated with a first-line standard of care therapy for gastroesophageal cancer, and the first line therapy was unsuccessful or the cancer subsequently relapsed. For example, where the first line standard of care therapy comprises fluorouracil chemotherapy and/or a platinum-based chemotherapy, the patients for selection typically receive a follow up PET and/or CT scan after 5-8 weeks following completion of the treatment. The scans may identify persistent (recurrent) local disease, unresectable locally advanced disease or metastatic disease. Subjects having received fluorouracil chemotherapy and/or a platinum-based chemotherapy in the first line standard of care therapy may be particularly suitable for treatment.

[0199] In addition, selected subjects are positive for HLA-A*02:01, HLA-A*02:03, or HLA-A*02:06, or another HLA-A*02 allele having the same protein sequence in the peptide binding domains. Patients who are HLA-A*02:05 positive are excluded from the study, because alloreactivity from ADP-A2M4 and from ADP-A2M4CD8 has previously been seen towards two HLA-A*02:05 positive cell lines. Patients with either HLA-A*02:07 or any A*02 null allele as the sole HLA-A*02 allele are also excluded due to decreased activity with these alleles.

[0200] Selected subjects also have a tumour that shows MAGE-A4 expression defined as 30% of tumour cells that are 2+ by immunohistochemistry (IHC).

[0201] Autologous cells are collected from selected subjects by leukapheresis for processing and manufacture into ADP-A2M4CD8. The heterologous TCR comprised in ADP-A2M4CD8 T cells comprises an alpha chain sequence comprised in SEQ ID NO: 2 and a beta chain sequence comprised in SEQ ID NO: 3. The heterologous CD8 co-receptor comprised in ADP-A2M4CD8 T cells comprises two CD8 alpha chains, each comprising an amino acid sequence of SEQ ID NO: 10. The surface-expressed heterologous TCR and surface-expressed heterologous CD8 co-receptor do not comprise the signal sequence.

[0202] A baseline tumour assessment is obtained prior to treatment. Then, subjects are administered (1) a second-line standard of care treatment (paclitaxel and ramucirumab) followed by ADP-A2M4CD8, or (2) ADP-A2M4CD8 followed by a second-line standard of care treatment (paclitaxel and ramucirumab). Lymphodepleting chemotherapy using fludarabine and cyclophosphamide (Day-7 through Day-4 pre-ADP-A2M4CD8) is provided in anticipation of administration of ADP-A2M4CD8. Subjects are optionally administered nivolumab after protocol (1) or (2) above.

[0203] For subjects on protocol (1) ramucirumab may be administered on day 1 and 15, and paclitaxel on days 1, 8 and 15. Leukapheresis may be conducted between days 26 and 28. A second cycle of ramucirumab and paclitaxel may be administered between days 29 and 56. A third cycle of ramucirumab and paclitaxel may be administered between days 57 and 84. Lymphodepletion may begin on day 85. ADP-A2M4CD8 may be infused on day 92. If leukapheresis cannot be performed at the end of the first cycle of ramucirumab and paclitaxel then it may instead be performed at the end of the second cycle. ADP-A2M4CD8 may then be infused either after a fourth cycle of ramucirumab and paclitaxel, or as soon as ADP-A2M4CD8 are available after then third cycle of ramucirumab and paclitaxel.

[0204] Between 110.sup.8 to 110.sup.10 ADP-A2M4CD8 T cells are administered to the subjects. The initial dose selected for ADP-A2M4CD8 is 110.sup.9 transduced cells (Range: 0.810.sup.9-1.210.sup.9 transduced cells). Administration is via single intravenous infusion. Subjects are monitored immediately post-infusion monitoring (Day 1 through Day 8). Subjects are monitored weekly until Week 4 post-infusion. Then, subjects are monitored at Weeks 6, 8, 12, 16, and 24 and at least every 3 months thereafter until disease progression. Tumour response is assessed according to response evaluation criteria in solid tumours (RECIST) v1.1. Additional data collected includes: [0205] Core needle biopsies, to directly evaluate the immune landscape inside the tumour at baseline and during the course of the study. [0206] Cytokine levels in the serum at baseline and during the course of the study. [0207] Humoral immune responses to tumour antigens at baseline and during the course of the study, using serum. [0208] Antibodies to ADP-A2M4CD8 at baseline and during the course of the study, using serum. [0209] Soluble markers representing the tumour and its microenvironment, using liquid biopsies. For instance, markers of circulating tumour cells (CTCs), exosome, and cell-free DNA (cfDNA) produced by dying tumour cells) may be used to monitor both the molecular signature of the tumour burden (including the expression of the target antigen) and the immune response. The analysis of such soluble markers allows estimation and genetic profiling of the global tumour burden, including expression of MAGE-A4 mRNA and mutational profiling. The analysis of such soluble markers also allows systemic assessment of the immune response. [0210] The phenotype and activity of the gene-modified T cells before and after infusion. The relevant assays may be performed using blood and, if resection is performed, tumour. The assays include: (i) phenotype analysis for determination of T-cell lineages in cell product and in the blood (and, if resection performed, tumour) post-infusion; (ii) quantitation of the senescence and activation status of immune subsets from PBMCs; (iii) analysis of gene expression or epigenetic profile to reflect phenotype and functional state of the cells; and/or (iv) direct functional assessment of the cells. [0211] Persistence of infused engineered cells, and correlation with therapeutic effect. Persistence may be determined by the copies of gene-modified DNA per g DNA, and/or data on the number of transduced cells per L or relative to total lymphocyte number. Well-established methodologies include (i) quantitation of ADP-A2M4CD8 cells by quantitative PCR of transgene from DNA extracted from frozen PBMCs, and (ii) quantitation and phenotyping of ADP-A2M4CD8 cells by flow cytometry, DNA and RNA analysis from frozen PBMCs.

[0212] Doses of up to 1010.sup.9 ADP-A2M4CD8 have been administered and shown to be well-tolerated. Emerging data indicates that treatment outcomes may be improved when second-line treatment of gastroesophageal cancer comprises administration of ADP-A2M4CD8. Outcomes may, for instance, be improved relative to a standard-of care second-line treatment of relapsed gastroesophageal cancer, such as ramucirumab in combination with paclitaxel (and optionally nivolumab).

Example 2. Third Line Treatment of Gastroesophageal Cancer

[0213] Subjects are selected for treatment with ADP-A2M4CD8. In brief, subjects eligible for selection must have been previously treated with a first line standard of care therapy and a second-line standard of care therapy for gastroesophageal cancer, and the first- and second-line therapies were unsuccessful or the cancer subsequently relapsed. For example, the cancer may be relapsed esophageal squamous cell carcinoma.

[0214] Selected subjects are also positive for HLA-A*02:01, HLA-A*02:03, or HLA-A*02:06, or another HLA-A*02 allele having the same protein sequence in the peptide binding domains. Patients who are HLA-A*02:05 positive are excluded from the study, because alloreactivity from ADP-A2M4 and from ADP-A2M4CD8 has previously been seen towards two HLA-A*02:05 positive cell lines. Patients with either HLA-A*02:07 or any A*02 null allele as the sole HLA-A*02 allele are also excluded due to decreased activity with these alleles. In addition, selected subjects have a tumour that shows MAGE-A4 expression defined as 30% of tumour cells that are 2+ by immunohistochemistry (IHC).

[0215] Autologous cells are collected from enrolled subjects by leukapheresis for processing and manufacture into ADP-A2M4CD8. The heterologous TCR comprised in ADP-A2M4CD8 T cells comprises an alpha chain sequence comprised in SEQ ID NO: 2 and a beta chain sequence comprised in SEQ ID NO: 3. The heterologous CD8 co-receptor comprised in ADP-A2M4CD8 T cells comprises two CD8 alpha chains, each comprising an amino acid sequence of SEQ ID NO: 10. The surface-expressed heterologous TCR and surface-expressed heterologous CD8 co-receptor do not comprise the signal sequence.

[0216] A baseline tumour assessment was obtained prior to treatment. Then, the subjects are administered ADP-A2M4CD8 a third-line standard-of-care treatment, typically a PD-1 axis binding antagonist such as pembrolizumab. Lymphodepleting chemotherapy using fludarabine and cyclophosphamide (Day-7 through Day-4 pre-ADP-A2M4CD8) is provided in anticipation of administration of ADP-A2M4CD8.

[0217] Between 110.sup.8 to 110.sup.10 ADP-A2M4CD8 T cells are administered to the subjects. The initial dose selected for ADP-A2M4CD8 is 110.sup.9 transduced cells (Range: 0.810.sup.9-1.210.sup.9 transduced cells). Administration is via single intravenous infusion.

[0218] Subjects are monitored immediately post-infusion monitoring (Day 1 through Day 8). Subjects were monitored weekly until Week 4 post-infusion. Then, subjects are monitored at Weeks 6, 8, 12, 16, and 24 and at least every 3 months thereafter until disease progression. Tumour response is assessed according to response evaluation criteria in solid tumours (RECIST) v1.1. Additional data collected includes: [0219] Core needle biopsies, to directly evaluate the immune landscape inside the tumour at baseline and during the course of the study. [0220] Cytokine levels in the serum at baseline and during the course of the study. [0221] Humoral immune responses to tumour antigens at baseline and during the course of the study, using serum. [0222] Antibodies to ADP-A2M4CD8 at baseline and during the course of the study, using serum. [0223] Soluble markers representing the tumour and its microenvironment, using liquid biopsies. For instance, markers of circulating tumour cells (CTCs), exosome, and cell-free DNA (cfDNA) produced by dying tumour cells) may be used to monitor both the molecular signature of the tumour burden (including the expression of the target antigen) and the immune response. The analysis of such soluble markers allows estimation and genetic profiling of the global tumour burden, including expression of MAGE-A4 mRNA and mutational profiling. The analysis of such soluble markers also allows systemic assessment of the immune response. [0224] The phenotype and activity of the gene-modified T cells before and after infusion. The relevant assays may be performed using blood and, if resection is performed, tumour. The assays include: (i) phenotype analysis for determination of T-cell lineages in cell product and in the blood (and, if resection performed, tumour) post-infusion; (ii) quantitation of the senescence and activation status of immune subsets from PBMCs; (iii) analysis of gene expression or epigenetic profile to reflect phenotype and functional state of the cells; and/or (iv) direct functional assessment of the cells. [0225] Persistence of infused engineered cells, and correlation with therapeutic effect. Persistence may be determined by the copies of gene-modified DNA per g DNA, and/or data on the number of transduced cells per L or relative to total lymphocyte number. Well-established methodologies include (i) quantitation of ADP-A2M4CD8 cells by quantitative PCR of transgene from DNA extracted from frozen PBMCs, and (ii) quantitation and phenotyping of ADP-A2M4CD8 cells by flow cytometry, DNA and RNA analysis from frozen PBMCs.

[0226] Doses of up to 1010.sup.9 ADP-A2M4CD8 have been administered and shown to be well-tolerated. Emerging data indicates that treatment outcomes may be improved when third-line treatment of gastroesophageal cancer comprises administration of ADP-A2M4CD8. Outcomes may, for instance, be improved relative to a standard-of care third-line treatment of gastroesophageal cancer, such as PD-1 axis binding antagonist monotherapy.

Example 3. First-Line Treatment of Gastroesophageal Cancer

[0227] Subjects are selected for treatment with ADP-A2M4CD8. In brief, subjects eligible for selection must (A) have relapsed following curative intent treatment for locally advanced cancer (B) have received a first diagnosis of unresectable locally advanced cancer or metastatic cancer.

[0228] In addition, selected subjects are positive for HLA-A*02:01, HLA-A*02:03, or HLA-A*02:06, or another HLA-A*02 allele having the same protein sequence in the peptide binding domains. Patients who are HLA-A*02:05 positive are excluded from the study, because alloreactivity from ADP-A2M4 and from ADP-A2M4CD8 has previously been seen towards two HLA-A*02:05 positive cell lines. Patients with either HLA-A*02:07 or any A*02 null allele as the sole HLA-A*02 allele are also excluded due to decreased activity with these alleles.

[0229] Selected subjects also have a tumour that shows MAGE-A4 expression defined as 30% of tumour cells that are 2+ by immunohistochemistry (IHC).

[0230] Autologous cells are collected from selected subjects by leukapheresis for processing and manufacture into ADP-A2M4CD8. The heterologous TCR comprised in ADP-A2M4CD8 T cells comprises an alpha chain sequence comprised in SEQ ID NO: 2 and a beta chain sequence comprised in SEQ ID NO: 3. The heterologous CD8 co-receptor comprised in ADP-A2M4CD8 T cells comprises two CD8 alpha chains, each comprising an amino acid sequence of SEQ ID NO: 10. The surface-expressed heterologous TCR and surface-expressed heterologous CD8 co-receptor do not comprise the signal sequence.

[0231] A baseline tumour assessment is obtained prior to treatment. Then, subjects are administered an oxaliplatin-based regimen followed by ADP-A2M4CD8. Subjects may additionally be administered (i) PD-1 axis binding antagonist such a nivolumab or pembrolizumab and/or (ii) fluorouracil (5FU). Lymphodepleting chemotherapy using fludarabine and cyclophosphamide (Day-7 through Day-4 pre-ADP-A2M4CD8) is provided in anticipation of administration of ADP-A2M4CD8.

[0232] Between 110.sup.8 to 110.sup.10 ADP-A2M4CD8 T cells are administered to the subjects. The initial dose selected for ADP-A2M4CD8 is 110.sup.9 transduced cells (Range: 0.810.sup.9-1.210.sup.9 transduced cells). Administration is via single intravenous infusion.

[0233] Subjects are monitored immediately post-infusion monitoring (Day 1 through Day 8). Subjects are monitored weekly until Week 4 post-infusion. Then, subjects are monitored at Weeks 6, 8, 12, 16, and 24 and at least every 3 months thereafter until disease progression. Tumour response is assessed according to response evaluation criteria in solid tumours (RECIST) v1.1. Additional data collected includes: [0234] Core needle biopsies, to directly evaluate the immune landscape inside the tumour at baseline and during the course of the study. [0235] Cytokine levels in the serum at baseline and during the course of the study. [0236] Humoral immune responses to tumour antigens at baseline and during the course of the study, using serum. [0237] Antibodies to ADP-A2M4CD8 at baseline and during the course of the study, using serum. [0238] Soluble markers representing the tumour and its microenvironment, using liquid biopsies. For instance, markers of circulating tumour cells (CTCs), exosome, and cell-free DNA (cfDNA) produced by dying tumour cells) may be used to monitor both the molecular signature of the tumour burden (including the expression of the target antigen) and the immune response. The analysis of such soluble markers allows estimation and genetic profiling of the global tumour burden, including expression of MAGE-A4 mRNA and mutational profiling. The analysis of such soluble markers also allows systemic assessment of the immune response. [0239] The phenotype and activity of the gene-modified T cells before and after infusion. The relevant assays may be performed using blood and, if resection is performed, tumour. The assays include: (i) phenotype analysis for determination of T-cell lineages in cell product and in the blood (and, if resection performed, tumour) post-infusion; (ii) quantitation of the senescence and activation status of immune subsets from PBMCs; (iii) analysis of gene expression or epigenetic profile to reflect phenotype and functional state of the cells; and/or (iv) direct functional assessment of the cells. [0240] Persistence of infused engineered cells, and correlation with therapeutic effect. Persistence may be determined by the copies of gene-modified DNA per g DNA, and/or data on the number of transduced cells per L or relative to total lymphocyte number. Well-established methodologies include (i) quantitation of ADP-A2M4CD8 cells by quantitative PCR of transgene from DNA extracted from frozen PBMCs, and (ii) quantitation and phenotyping of ADP-A2M4CD8 cells by flow cytometry, DNA and RNA analysis from frozen PBMCs.

[0241] Doses of up to 1010.sup.9 ADP-A2M4CD8 have been administered and shown to be well-tolerated. Emerging data indicates that treatment outcomes may be improved when a first-line treatment of gastroesophageal cancer comprises administration of ADP-A2M4CD8. Outcomes may, for instance, be improved relative to a standard-of care first-line treatment, such as an oxaliplatin based regimen.

Example 4. Efficacy Data from the Phase I SURPASS Trial of ADP-A2M4CD8, a Next Generation T-Cell Receptor T-Cell Therapy, in Patients with Advanced Esophageal, Esophagogastric Junction, or Gastric Cancer

[0242] Autologous T-cells were obtained by leukapheresis, transduced with a self-inactivating lentiviral vector expressing the MAGE-A4-specific TCR and the CD8 co-receptor, and infused back to the patients as ADP-A2M4CD8 following lymphodepleting chemotherapy.

[0243] Fifteen patients with MAGE-A4-positive esophageal (n=3), esophagogastric junction (EGJ; n=10) and gastic (n=2) cancer were treated with ADP-A2M4CD8. Of these, three (EGJ n=2, gastric n=1) were in combination with nivolumab. Prior therapies included chemotherapy (n=15), PD-(L) 1 inhibitors (n=9, 60%), anti-VEGFR (n=9, 60% and anti-HER2 (n=4, 27%). All but one of the patients have liver, peritoneal and/or retroperitoneal metastases. The baseline characteristics of the patients are provided in the table below:

TABLE-US-00001 Nivolumab combination Monotherapy Characteristic (n = 3) (n = 12) Sex, n (%) Male 3 (100.0) 10 (83.3) Female 0 2 (16.7) Age, years, median (min, max) 59.0 (53, 73) 55.5 (31, 71) H-score.sup.a, median (min, max) 265 (145, 295) 227.5 (160, 300) ECOG at baseline, n (%) 0 1 (33.3) 1 (8.3) 1 2 (66.7) 11 (91.7) Bridging therapy, yes, n (%) 3 (100.0) 9 (75.0) Transduced cells, 10.sup.9 cells, 4.07, 7.95 1.02, 9.90 min, max Prior lines of systemic therapy, 2.0 (1, 2) 3.0 (1, 5) median (min, max) SLD 50 mm, n (%) 0 10 (83.3) .sup.aH-score: 1 (% of 1+ cells) + 2 (% of 2+ cells) + 3 (% of 3+ cells). ECOG, Eastern Cooperative Oncology Group; max, maximum; min, minimum; SLD, sum of the longest diameters of target lesions; EGJ, esophagogastric junction.

[0244] The overall response rate (ORR) per Response Evaluation Criteria in Solid Tumours (RECIST) v1.1 by investigator review was 20.0% (3 partial responses; FIG. 6).

[0245] The disease control rate was 80.0% (3 partial responses and nine stable disease).

[0246] The duration of the response ranged from 5.0 to 29.3 weeks. The data shown in FIG. 6 indicates changes from baseline SLD through progression of disease or prior to surgical resection.

SEQUENCE LISTING

TABLE-US-00002 MAGE-A4230-239 SEQIDNO:1 GVYDGREHTV MAGE-A4TCRalphachain(CDRsboldunderlined,signal sequenceitalicunderlined) SEQIDNO:2 MKKHLTTFLVILWLYFYRGNGKNQVEQSPQSLIILEGKNCTLQCNYTVSPFSNLRWYKQDT GRGPVSLTILTFSENTKSNGRYTATLDADTKQSSLHITASQLSDSASYICVVSGGTDSWGK LQFGAGTQVVVTPDIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKT VLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTN LNFQNLSVIGFRILLLKVAGENLLMTLRLWSSGSRAKR MAGE-A4TCRbetachain(CDRsboldunderlined,signalsequence italicunderlined) SEQIDNO:3 MASLLFFCGAFYLLGTGSMDADVTQTPRNRITKTGKRIMLECSQTKGHDRMYWYRQDPGLG LRLIYYSFDVKDINKGEISDGYSVSRQAQAKFSLSLESAIPNQTALYFCATSGQGAYEEQF FGPGTRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKE VHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQD RAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVK RKDSRG MAGE-A4TCRalphachainCDR1 SEQIDNO:4 VSPFSN MAGE-A4TCRalphachainCDR2 SEQIDNO:5 LTFSEN MAGE-A4TCRalphachainCDR3 SEQIDNO:6 CVVSGGTDSWGKLQF MAGE-A4TCRbetachainCDR1 SEQIDNO:7 KGHDR MAGE-A4TCRbetachainCDR2 SEQIDNO:8 SFDVKD MAGE-A4TCRbetachainCDR3 SEQIDNO:9 CATSGQGAYEEQFF CD8alphachain(CDR-likeloopsboldunderlined, signalsequenceitalicunderlined) SEQIDNO:10 MALPVTALLLPLALLLHAARPSQFRVSPLDRTWNLGETVELKCQVLLSNPTSGCSWLFQPR GAAASPTFLLYLSQNKPKAAEGLDTQRFSGKRLGDTFVLTLSDFRRENEGYYFCSALSNSI MYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDI YIWAPLAGTCGVLLLSLVITLYCNHRNRRRVCKCPRPVVKSGDKPSLSARYV CD8alphachainCDR1 SEQIDNO:11 VLLSNPTSG CD8alphachainCDR2 SEQIDNO:12 YLSQNKPK CD8alphachainCDR3 SEQIDNO:13 LSNSIM PD1-HumanProgrammedcelldeathprotein(Homosapiens) SEQIDNO:14 MQIPQAPWPVVWAVLQLGWRPGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSESNTS ESFVLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGRDFHMSVVRARRNDSGT YLCGAISLAPKAQIKESLRAELRVTERRAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGS LVLLVWVLAVICSRAARGTIGARRTGQPLKEDPSAVPVFSVDYGELDFQWREKTPEPPVP CVPEQTEYATIVEPSGMGTSSPARRGSADGPRSAQPLRPEDGHCSWPL PD1L1-HumanProgrammedcelldeath1ligand1(Homosapiens) SEQIDNO:15 MRIFAVFIFMTYWHLLNAFTVTVPKDLYVVEYGSNMTIECKFPVEKQLDLAALIVYWEME DKNIIQFVHGEEDLKVQHSSYRQRARLLKDQLSLGNAALQITDVKLQDAGVYRCMISYGG ADYKRITVKVNAPYNKINQRILVVDPVTSEHELTCQAEGYPKAEVIWTSSDHQVLSGKTT TTNSKREEKLFNVTSTLRINTTTNEIFYCTFRRLDPEENHTAELVIPELPLAHPPNERTH LVILGAILLCLGVALTFIFRLRKGRMMDVKKCGIQDTNSKKQSDTHLEET PD1L2-HumanProgrammedcelldeath1ligand2(Homosapiens) SEQIDNO:16 MIFLLLMLSLELQLHQIAALFTVTVPKELYIIEHGSNVTLECNEDTGSHVNLGAITASLQ KVENDTSPHRERATLLEEQLPLGKASFHIPQVQVRDEGQYQCIIIYGVAWDYKYLTLKVK ASYRKINTHILKVPETDEVELTCQATGYPLAEVSWPNVSVPANTSHSRTPEGLYQVTSVL RLKPPPGRNFSCVFWNTHVRELTLASIDLQSQMEPRTHPTWLLHIFIPFCIIAFIFIATV IALRKQLCQKLYSSKDTTKRPVTTTKREVNSAI NivolumabHeavyChainSequence SEQIDNO:17 QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRYY ADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVTVSSASTKGPS VFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKP KDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVESCSV MHEALHNHYTQKSLSLSLGK NivolumabLightChainSequence SEQIDNO:18 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPA RFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQGTKVEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSENRGEC PembrolizumabHeavyChainSequence SEQIDNO:19 QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQGLEWMGGINPSNGGTNE NEKFKNRVTLTTDSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMGEDYWGQGTTVTVSS ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSV FLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQENSTY RVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEG NVFSCSVMHEALHNHYTQKSLSLSLGK PembrolizumabLightChainSequence SEQIDNO:20 EIVLTQSPATLSLSPGERATLSCRASKGVSTSGYSYLHWYQQKPGQAPRLLIYLASYLES GVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQHSRDLPLTFGGGTKVEIKRTVAAPSVE IFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS STLTLSKADYEKHKVYACEVTHQGLSSPVTKSENRGEC CemiplimabHeavyChainSequence SEQIDNO:21 EVQLLESGGVLVQPGGSLRLSCAASGFTFSNFGMTWVRQAPGKGLEWVSGISGGGRDTYF ADSVKGRFTISRDNSKNTLYLQMNSLKGEDTAVYYCVKWGNIYFDYWGQGTLVTVSSAST KGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY SLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVELE PPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQENSTYRVV SVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQV SLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVE SCSVMHEALHNHYTQKSLSLSLGK CemiplimabLightChainSequence SEQIDNO:22 DIQMTQSPSSLSASVGDSITITCRASLSINTELNWYQQKPGKAPNLLIYAASSLHGGVPS RFSGSGSGTDFTLTIRTLQPEDFATYYCQQSSNTPFTFGPGTVVDERRTVAAPSVFIEPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSENRGEC DurvalumabHeavyChainSequence SEQIDNO:23 EVQLVESGGGLVQPGGSLRLSCAASGFTFSRYWMSWVRQAPGKGLEWVANIKQDGSEKYY VDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREGGWFGELAFDYWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEFEG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKENWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPSRE EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK DurvalumabLightChainSequence SEQIDNO:24 EIVLTQSPGTLSLSPGERATLSCRASQRVSSSYLAWYQQKPGQAPRLLIYDASSRATGIP DRFSGSGSGTDETLTISRLEPEDFAVYYCQQYGSLPWTFGQGTKVEIKRTVAAPSVFIFP PSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSENRGEC AtezolizumabHeavyChainSequence SEQIDNO:25 EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYY ADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSAS TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPS VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKENWYVDGVEVHNAKTKPREEQYAST YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ GNVFSCSVMHEALHNHYTQKSLSLSPGK AtezolizumabLightChainSequence SEQIDNO:26 DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPS RFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLINNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSENRGEC AvelumabHeavyChainSequence SEQIDNO:27 EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYIMMWVRQAPGKGLEWVSSIYPSGGITFY ADTVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARIKLGTVTTVDYWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGG PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKENWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDE LTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK AvelumabLightChainSequence SEQIDNO:28 QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYDVSNRPSGV SNRFSGSKSGNTASLTISGLQAEDEADYYCSSYTSSSTRVFGTGTKVTVLGQPKANPTVT LFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASS YLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS MDX1105HeavyChainSequence SEQIDNO:29 QVQLVQSGAEVKKPGSSVKVSCKTSGDTESTYAISWVRQAPGQGLEWMGGIIPIFGKAHYAQKFQG RVTITADESTSTAYMELSSLRSEDTAVYFCARKFHFVSGSPFGMDVWGQGTTVTVSSASTKGPSVE PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTQTYICNVNHKPSNTKVDKKVEPKSCDKT MDX1105LightChainSequence SEQIDNO:30 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSG SGTDFTLTISSLEPEDFAVYYCQQRSNWPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVV CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSENRGEC DostarlimabHeavyChainSequence SEQIDNO:31 EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVSTISGGGSYTYYQDSVKG RFTISRDNSKNTLYLQMNSLRAEDTAVYYCASPYYAMDYWGQGTTVTVSSASTKGPSVFPLAPCSR STSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYT CNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQ EDPEVQFNWYVDGVEVHNAKTKPREEQENSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEK TISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS DGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK DostarlimabLightChainSequence SEQIDNO:32 DIQLTQSPSELSAYVGDRVTITCKASQDVGTAVAWYQQKPGKAPKLLIYWASTLHTGVPSRFSGSG SGTEFTLTISSLQPEDFATYYCQHYSSYPWTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASV VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSENRGEC