Dosage regimen for combination therapy consisting of an engineered T cell and a PD-1 axis binding antagonist

Abstract

The present disclosure relates to a method of treating cancer and a population of modified T cells for use in a method of treating cancer.

Description

Technical Field

[0001] The present disclosure relates to a method of treating cancer, a population of modified T cells for use in a method of treating cancer, and a PD-1 axis-binding antagonist for use in a method of treating cancer.

Background Art

[0002] Immunotherapy is an important component of the anti-cancer toolkit. Immunological effectors such as anti-tumor monoclonal antibodies, T cells expressing chimeric antigen receptors (CAR T cells), and TCR-engineered T cells can be adoptively transferred into an individual to promote an anti-cancer immune response and thereby treat the disease.

[0003] However, the therapeutic ability of immunotherapy can be limited by the ability of cancer cells to regulate the immune response. To effectively eliminate tumors, it is important to maintain an activated and persistent T cell response. However, solid tumors have an immunosuppressive tumor microenvironment that is promoted by the cancer cells themselves as well as by the infiltration of suppressive immune cells such as myeloid-derived suppressor cells (MDSC) and regulatory T cells (Treg). The immunosuppressive microenvironment can suppress the anti-tumor T cell response and / or promote T cell exhaustion, making it difficult to maintain an activated T cell response and eliminate tumors.

[0004] Therefore, in order to overcome the immunosuppressive tumor microenvironment, it is necessary to develop a dosing regimen for T cells for immunotherapy that promotes the survival and function of T cells.

Summary of the Invention

[0005] The inventors have identified that an improved anti-tumor immune response can be obtained by administering T cells for tumor-specific immunotherapy in combination with a PD-1 axis-binding antagonist. In particular, the inventors propose that administration of a PD-1 axis-binding antagonist can sustain the activity of endogenous T cells present in the immunosuppressive tumor microenvironment. The inventors further propose that such administration helps maintain the function of adoptively transferred tumor-specific T cells, including their ability to migrate into memory T cells. In this way, the therapeutic effect of adoptively transferred tumor-specific T cells can be enhanced and / or extended. The inventors have further identified an optimal dosing schedule for such combination therapy.

[0006] Accordingly, the present disclosure provides a method of treating cancer in an individual, comprising (a) administering to the individual a population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous TCR capable of binding to MAGE-A4, and (b) administering to the individual a first dose of a PD-1 axis-binding antagonist about 3 to about 5 weeks after step (a), which comprises maintaining in the individual (i) the population of modified T cells and / or (ii) the function of endogenous T cells. The present disclosure also provides the following - A population of modified T cells for use in a method of treating cancer in an individual, wherein the modified T cells comprise a heterologous CD8 co-receptor and a heterologous TCR capable of binding to MAGE-A4, and the method comprises (a) administering the population to the individual, and (b) administering to the individual a first dose of a PD-1 axis-binding antagonist about 3 to about 5 weeks after step (a), which comprises maintaining in the individual (i) the population of modified T cells and / or (ii) the function of endogenous T cells. A PD-1 axis-binding antagonist for use in a method of treating cancer in an individual, the method comprising: (a) administering to the individual a population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous TCR capable of binding to MAGE-A4; and (b) administering to the individual a first dose of the PD-1 axis-binding antagonist about 3 to about 5 weeks after step (a), thereby maintaining the function of (i) the population of modified T cells and / or (ii) endogenous T cells in the individual. BRIEF DESCRIPTION OF THE DRAWINGS

[0007]

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[0008] It should be understood that the different applications of the disclosed methods and products can be adjusted according to specific needs in the art. It should also be understood that the terms used herein are for the purpose of describing only particular embodiments of the present disclosure and are not intended to be limiting.

[0009] All publications, patents, and patent applications cited herein, whether supra or infra, are hereby incorporated by reference in their entirety.

[0010] General Definitions Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0011] As used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural references unless the context 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.

[0012] Generally, 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 includes the step of administering such a population, but the method may include additional steps, such as the step of administering a further therapeutic agent.

[0013] In some aspects of the present 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 includes the step of administering such a population and does not include additional steps.

[0014] The terms "protein" and "polypeptide" are used interchangeably herein and are intended to refer to a polymer chain of amino acids of any length.

[0015] For the purposes of the present disclosure, 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 into the first sequence for optimal alignment with the second sequence). The nucleotide residues at corresponding nucleotide positions are then compared. If a position in the first sequence is occupied by the same nucleotide residue as the corresponding position in the second sequence, the nucleotides are identical at that position. The percent identity between 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 sequence × 100).

[0016] Typically, sequence comparisons are performed over the length of a reference sequence. For example, if a user desires to determine whether a given (“test”) sequence has a particular percent identity to sequence number X, then sequence number X is the reference sequence. For example, to evaluate whether a sequence is at least 80% identical to sequence number X (an example of a reference sequence), one of ordinary skill in the art aligns over the length of sequence number X and identifies how many positions in the test sequence are identical to the positions in sequence number X. If at least 80% of the positions are identical, then the test sequence is at least 80% identical to sequence number X. If the sequence is shorter than sequence number X, gap or missing positions should be considered non-identical positions.

[0017] One of ordinary skill in the art is aware of different computer programs available for determining homology or identity between two sequences. For example, sequence comparison and determination of the percent identity between two sequences can be accomplished using mathematical algorithms.

[0018] Treatment of cancer The present disclosure provides a method of treating cancer in an individual, comprising (a) administering to the individual a population of modified T cells comprising a heterologous CD8 coreceptor and a heterologous TCR capable of binding to MAGE-A4, and (b) administering to the individual a first dose of a PD-1 axis-binding antagonist about 3 to about 5 weeks after step (a), thereby maintaining the function of (i) the population of modified T cells and / or (ii) endogenous T cells in the individual. The PD-1 axis-binding antagonist may alternatively be known as a PD-PD-L1-PD-L2 axis inhibitor.

[0019] The present disclosure also provides a population of modified T cells for use in a method of treating cancer in an individual, the modified T cells comprising a heterologous CD8 coreceptor and a heterologous TCR capable of binding to MAGE-A4, the method comprising (a) administering the population to the individual, and (b) maintaining in the individual (i) the population of modified T cells and / or (ii) the function of endogenous T cells by administering a first dose of a PD-1 axis-binding antagonist to the individual about 3 to about 5 weeks after step (a). Additionally, the present disclosure provides the use of a population of modified T cells in the manufacture of a medicament for use in a method of treating cancer in an individual, the modified T cells comprising a heterologous CD8 coreceptor and a heterologous TCR capable of binding to MAGE-A4, the method comprising (a) administering the population to the individual, and (b) maintaining in the individual (i) the population of modified T cells and / or (ii) the function of endogenous T cells by administering a first dose of a PD-1 axis-binding antagonist to the individual about 3 to about 5 weeks after step (a).

[0020] The present disclosure further provides a PD-1 axis-binding antagonist for use in a method of treating cancer in an individual, the method comprising (a) administering to the individual a population of modified T cells comprising a heterologous CD8 coreceptor and a heterologous TCR capable of binding to MAGE-A4, and (b) maintaining in the individual (i) the population of modified T cells and / or (ii) the function of endogenous T cells by administering a first dose of a PD-1 axis-binding antagonist to the individual about 3 to about 5 weeks after step (a). Additionally, the present disclosure provides the use of a PD-1 axis-binding antagonist in the manufacture of a medicament for use in a method of treating cancer in an individual, the method comprising (a) administering to the individual a population of modified T cells comprising a heterologous CD8 coreceptor and a heterologous TCR capable of binding to MAGE-A4, and (b) maintaining in the individual (i) the population of modified T cells and / or (ii) the function of endogenous T cells by administering a first dose of a PD-1 axis-binding antagonist to the individual about 3 to about 5 weeks after step (a).

[0021] Treatment of Cancer in an Individual The present disclosure relates to the administration of (i) a population of modified T cells comprising a heterologous CD8 coreceptor and a heterologous TCR capable of binding to MAGE-A4, and (ii) a PD-1 axis binding antagonist for treating cancer in an individual. The individual may be, for example, a human. The individual may be, for example, a non-human mammal such as a dog, a cat or a horse.

[0022] The heterologous TCR comprised in the modified T cells is capable of binding to MAGE-A4. MAGE-A4 is a well-known cancer antigen with restricted expression in normal (i.e., non-cancerous) tissues. MAGE-A4 has been shown to suppress p53 targets (such as BAX and CDKN1A), and is a binding partner of the oncogene gankyrin. The heterologous TCR may bind, for example, to the peptide sequence GVYDGREHTV (SEQ ID NO: 1) known as MAGE-A4230-239 comprised in MAGE-A4. The heterologous TCR may bind, for example, to a complex comprising MAGE-A4 (such as SEQ ID NO: 1) and an HLA molecule such as * HLA-A02.

[0023] Accordingly, the cancer to be treated may be a cancer that expresses MAGE-A4. MAGE-A4 expression has been reported for many types of cancer. The cancer may be, for example, a solid tumor. The cancer may be, for example, urothelial cancer, head and neck cancer, non-small cell lung cancer (NSCLC), esophageal cancer, esophagogastric junction cancer, gastric cancer, ovarian cancer, melanoma, or endometrial cancer.

[0024] Accordingly, the cancer to be treated may be ovarian cancer. The term "ovarian cancer" is used herein to describe cancers that begin in the cells of the ovary, fallopian tube, or peritoneum. The term "ovarian cancer" includes epithelial cancers, and epithelial tumors include serous, endometrioid, clear cell, mucinous, mixed tumors, and Brenner tumors. The term "ovarian cancer" also includes germ cell malignancies, sex cord-stromal tumors, and fallopian tube cancers. The term "ovarian cancer" includes primary and metastatic ovarian cancers. The term "ovarian cancer" includes ovarian cancers that have recurred or are refractory. The term "ovarian cancer" includes ovarian cancers that may have been previously treated but have recurred and / or have become partially sensitive, intolerant, or resistant to platinum-based therapies. Ovarian cancer may have been previously treated surgically. Ovarian cancer may have been previously treated with radiation therapy. Ovarian cancer may have been previously treated with chemotherapeutic agents such as doxorubicin, docetaxel, paclitaxel, nab-paclitaxel, ifosfamide, capecitabine, fluorouracil, bleomycin, etoposide, gemcitabine, cyclophosphamide, irinotecan, melphalan, pemetrexed, vinorelbine, topotecan, vincristine, vinblastine, or dactinomycin. Ovarian cancer may have been previously treated with platinum-based therapies such as carboplatin, cisplatin, or oxaliplatin.Ovarian cancer may have been previously treated with targeted therapies such as PARP inhibitors, anti-angiogenesis inhibitors, or PK inhibitors, and targeted therapies include bevacizumab, olaparib, niraparib, rucaparib, pazopanib, sorafenib, entrectinib, larotrectinib, trametinib, dabrafenib, vemurafenib, cobimetinib, milbecycliximab sorabtansine, offranergen obadenovec (VB-111), upifitamab rilsodotin (XMT-1536), batiraxcept (AVB-500), navicixizumab (OMP-305B83), oregovomab, nemvaleukin alpha (ALKS 4230), adavosertib (AZD1775), bezosertib (M6620, VX-970, VE-822), cediranib (AZD-2171), alpelisib (BYL719), tumor treating fields (TTFields), relacorilant (CORT125134), and PC14586. Ovarian cancer may have been previously treated with hormonal therapies such as anastrozole, exemestane, letrozole, leuprolide acetate, tamoxifen, megestrol acetate, or fulvestrant. Ovarian cancer may have been previously treated with any combination of the above treatments. Ovarian cancer can recur after becoming intolerant or resistant to platinum-based treatments. Ovarian cancer may progress after one or more cycles of platinum-based treatment, for example, ovarian cancer may progress after 1, 2, 3, 4, 5, 6, 7, or 8 cycles of platinum-based treatment, and ovarian cancer may progress within about 300 days after the administration of platinum-based treatment, for example, within 14 to 300 days, 21 to 270 days, 30 to 240 days, 60 to 210 days, 90 to 195 days after the administration of platinum-based treatment. Ovarian cancer may progress within 120 to 185 days after 1, 2, or 3 cycles of platinum-based treatment.

[0025] Population of modified T cells In each of the above-described aspects of the present disclosure, the method comprises administering to an individual a population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous TCR capable of binding to MAGE-A4. The T cells are "modified" by the presence of the heterologous CD8 co-receptor and the heterologous TCR. 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 can express the heterologous CD8 co-receptor and the heterologous TCR on their surface.

[0026] 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., does not naturally occur in that system. A "heterologous" polypeptide or nucleic acid can be introduced into the system by artificial or recombinant means. Thus, heterologous expression of a TCR can alter the specificity of a T cell. Heterologous expression of a CD8 co-receptor can confer functions associated with the CD8 co-receptor to a T cell. The heterologous CD8 co-receptor and the heterologous TCR are described in detail below.

[0027] The modified T cells can include CD4+ T cells. That is, the modified T cells can include T cells that express an endogenous CD4 co-receptor. The modified T cells can include CD8+ T cells. That is, the modified T cells can include T cells that express an endogenous CD8 co-receptor. The modified T cells can include CD4+ T cells and CD8+ T cells. That is, the modified T cells can include T cells that express an endogenous CD4 co-receptor and can include T cells that express an endogenous CD8 co-receptor. Both CD4+ T cells and CD8+ T cells can carry the heterologous CD8 co-receptor.

[0028] The modified T cells may be allogeneic to the individual. The modified T cells may preferably be autologous to the individual. In this case, the modified T cells can be produced by modifying endogenous cells obtained from the individual. Thus, the method may include producing a population. Methods for producing modified T cells are known in the art and are considered in the following examples. Typically, the modified T cells of the present disclosure are produced from cells such as peripheral blood mononuclear cells (PBMCs). T cells are typically selected from the collected cells and engineered to contain the desired modifications (here, a heterologous CD8 coreceptor and a heterologous TCR). Thus, the method may include (i) obtaining peripheral blood mononuclear cells (PBMCs) from an individual, (ii) selecting T cells from the PBMCs, and (iii) producing a population by modifying the selected T cells to express a heterologous CD8 coreceptor and a heterologous TCR capable of binding to MAGE-A4.

[0029] Heterologous TCR The modified T cells include a heterologous TCR capable of binding to MAGE-A4. In other words, the modified T cells express, for example, on their surface, a heterologous TCR capable of binding to MAGE-A4.

[0030] The heterologous TCR may be, for example, 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 be, for example, an affinity-enhanced TCR, such as a specific peptide-enhanced affinity receptor (SPEAR™) TCR.

[0031] The heterologous TCR can bind to MAGE-A4. The heterologous TCR can bind, for example, to GVYDGREHTV (SEQ ID NO: 1). The heterologous TCR can bind, for example, to MAGE-A4 (e.g., GVYDGREHTV (SEQ ID NO: 1)) and HLA-A *It can bind to a complex containing an HLA molecule such as 02. In any case, the binding may be specific. Specificity refers to the strength of the binding between a heterologous TCR and its target antigen. Specificity can be described by the dissociation constant Kd, that is, the ratio of the bound state to the unbound state of the receptor-ligand system. Typically, the fewer different antigens to which a heterologous TCR can bind other than MAGE-A4, the higher its binding specificity.

[0032] A heterologous TCR can bind to MAGE-A4 with a dissociation constant (Kd) of, for example, 0.01 μM to 100 μM, 0.01 μM to 50 μM, 0.01 μM to 20 μM, 10 μM to 1000 μM, 10 μM to 500 μM, or 50 μM to 500 μM. For example, in a preferred embodiment of the present disclosure, the heterologous TCR binds to MAGE-A4 with a Kd of 0.05 μM to 20.0 μM. For example, the heterologous TCR can bind to MAGE-A4 with a Kd of 0.01 μM, 0.02 μM, 0.03 μΜ, 0.04 μΜ, 0.05 μΜ, 0.06 μΜ, 0.07 μΜ, 0.08 μΜ, 0.09 μΜ, 0.1 μΜ, 0.15 μΜ, 0.2 μΜ, 0.25 μΜ, 0.3 μΜ, 0.35 μΜ, 0.4 μΜ, 0.45 μΜ, 0.5 μΜ, 0.55 μΜ, 0.6 μΜ, 0.65 μΜ, 0.7 μΜ, 0.75 μΜ, 0.8 μΜ, 0.85 μΜ, 0.9 μΜ, 0.95 μΜ, 1.0 μΜ, 1.5 μΜ, 2.0 μΜ, 2.5 μΜ, 3.0 μΜ, 3.5 μΜ, 4.0 μΜ, 4.5 μΜ, 5.0 μΜ, 5.5 μΜ, 6.0 μΜ, 6.5 μΜ, 7.0 μΜ, 7.5 μΜ, 8.0 μΜ, 8.5 μΜ, 9.0 μΜ, 9.5 μΜ, 10.0 μΜ, 20 μΜ, 30 μΜ, 40 μΜ, 50 μΜ, 60 μΜ, 70 μΜ, 80 μΜ, 90 μΜ, 100 μΜ, 150 μΜ, 200 μΜ, 250 μΜ, 300 μΜ, 350 μΜ, 400 μΜ, 450 μΜ, 500 μM. The Kd can be measured, for example, using surface plasmon resonance, optionally at 25°C, and optionally at a pH of 6.5 to 6.9 or 7.0 to 7.5. The dissociation constant Kd or koff / kon can be determined by experimentally measuring the dissociation rate constant koff and the association rate constant kon. The TCR dissociation constant can be measured using the soluble form of the TCR, which contains the TCRα chain variable domain and the TCRβ chain variable domain.

[0033] The heterologous TCR may include an α-chain variable domain having at least 80% sequence identity with the sequence of amino acid residues 22-125 of SEQ ID NO: 2, for example. The heterologous TCR may include a β-chain variable domain having at least 80% sequence identity with the sequence of amino acid residues 22-123 of SEQ ID NO: 3, for example. The heterologous TCR may include an α-chain variable domain having at least 80% sequence identity with the sequence of amino acid residues 22-125 of SEQ ID NO: 2, and a β-chain variable domain having at least 80% sequence identity with the sequence of amino acid residues 22-123 of SEQ ID NO: 3. The α-chain variable domain may have at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% sequence identity with the sequence of amino acid residues 22-125 of SEQ ID NO: 2, for example. The α-chain variable domain may comprise, or consist of, for example, amino acid residues 22-125 of SEQ ID NO: 2. The β-chain variable domain may have at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% sequence identity with the sequence of amino acid residues 22-123 of SEQ ID NO: 3, for example. The β-chain amino acid sequence may comprise, or consist of, for example, amino acid residues 22-123 of SEQ ID NO: 3.

[0034] A heterologous TCR may include an α-chain having at least 80% sequence identity with the sequence of amino acid residues 22 to 282 of SEQ ID NO: 2, for example. A heterologous TCR may include a β-chain having at least 80% sequence identity with the sequence of amino acid residues 22 to 311 of SEQ ID NO: 3, for example. A heterologous TCR may include an α-chain having at least 80% sequence identity with the sequence of amino acid residues 22 to 282 of SEQ ID NO: 2, and a variable domain of a β-chain having at least 80% sequence identity with the sequence of amino acid residues 22 to 311 of SEQ ID NO: 3. The α-chain variable domain may have at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% sequence identity with the sequence of amino acid residues 22 to 282 of SEQ ID NO: 2, for example. The α-chain variable domain may, for example, contain or consist of amino acid residues 22 to 282 of SEQ ID NO: 2. The β-chain variable domain may have at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% sequence identity with the sequence of amino acid residues 22 to 311 of SEQ ID NO: 3, for example. The β-chain amino acid sequence may, for example, contain or consist of amino acid residues 22 to 311 of SEQ ID NO: 3.

[0035] Heterologous TCRs are typically expressed with an N-terminal signal peptide that is cleaved prior to expression on the surface of T cells. In this regard, amino acids 1-21 of SEQ ID NO: 2 and SEQ ID NO: 3, respectively, are typically cleaved prior to the expression of the TCR on the surface of T cells. A heterologous TCR can include, for example, an α-chain amino acid sequence having at least 80% sequence identity with SEQ ID NO: 2. A heterologous TCR can include, for example, a β-chain amino acid sequence having at least 80% sequence identity with SEQ ID NO: 3. A heterologous TCR can include, for example, an α-chain amino acid sequence having at least 80% sequence identity with SEQ ID NO: 2 and a β-chain amino acid sequence having at least 80% sequence identity with SEQ ID NO: 3. The α-chain amino acid sequence can have, for example, at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% sequence identity with SEQ ID NO: 2. The α-chain amino acid sequence can, for example, comprise or consist of SEQ ID NO: 2. The β-chain amino acid sequence can have, for example, at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% sequence identity with SEQ ID NO: 3. The β-chain amino acid sequence can, for example, comprise or consist of SEQ ID NO: 3.

[0036] The heterologous TCR may include, for example, (i) an α-chain variable domain including a CDR1 including the sequence of SEQ ID NO: 4 or an amino acid sequence including one, two, or three amino acid insertions, deletions, or substitutions relative to the sequence of SEQ ID NO: 4, (ii) an α-chain variable domain including a CDR2 including the sequence of SEQ ID NO: 5 or an amino acid sequence including one, two, or three amino acid insertions, deletions, or substitutions relative to the sequence of SEQ ID NO: 5, (iii) an α-chain variable domain including a CDR3 including the sequence of SEQ ID NO: 6 or an amino acid sequence including one, two, or three amino acid insertions, deletions, or substitutions relative to the sequence of SEQ ID NO: 6, (iv) a β-chain variable domain including a CDR1 including the sequence of SEQ ID NO: 7 or an amino acid sequence including one, two, or three amino acid insertions, deletions, or substitutions relative to the sequence of SEQ ID NO: 7, (v) a β-chain variable domain including a CDR2 including the sequence of SEQ ID NO: 8 or an amino acid sequence including one, two, or three amino acid insertions, deletions, or substitutions relative to the sequence of SEQ ID NO: 8, and / or (vi) a β-chain variable domain including a CDR3 including the sequence of SEQ ID NO: 9 or an amino acid sequence including one, two, or three amino acid insertions, deletions, or substitutions relative to the sequence of SEQ ID NO: 9.

[0037] The α-chain of the heterologous TCR may include, for example, (i) an α-chain variable domain including a CDR1 including the sequence of SEQ ID NO: 4 or an amino acid sequence including one, two, or three amino acid insertions, deletions, or substitutions relative to the sequence of SEQ ID NO: 4, (ii) an α-chain variable domain including a CDR2 including the sequence of SEQ ID NO: 5 or an amino acid sequence including one, two, or three amino acid insertions, deletions, or substitutions relative to the sequence of SEQ ID NO: 5, and (iii) an α-chain variable domain including a CDR3 including the sequence of SEQ ID NO: 6 or an amino acid sequence including one, two, or three amino acid insertions, deletions, or substitutions relative to the sequence of SEQ ID NO: 6. The α-chain of the heterologous TCR may include, for example, (i) an α-chain variable domain including a CDR1 including the sequence of SEQ ID NO: 4, (ii) an α-chain variable domain including a CDR2 including the sequence of SEQ ID NO: 5, and (iii) an α-chain variable domain including a CDR3 including the sequence of SEQ ID NO: 6.

[0038] The β-chain of the heterologous TCR may include, for example, a β-chain variable domain including a CDR1 including the sequence of SEQ ID NO: 7, or an amino acid sequence including an insertion, deletion or substitution of 1, 2 or 3 amino acids relative to the sequence of SEQ ID NO: 7; a β-chain variable domain including a CDR2 including the sequence of SEQ ID NO: 8, or an amino acid sequence including an insertion, deletion or substitution of 1, 2 or 3 amino acids relative to the sequence of SEQ ID NO: 8; and a β-chain variable domain including a CDR3 including the sequence of SEQ ID NO: 9, or an amino acid sequence including an insertion, deletion or substitution of 1, 2 or 3 amino acids relative to the sequence of SEQ ID NO: 9. The β-chain of the heterologous TCR may include, for example, a β-chain variable domain including a CDR1 including the sequence of SEQ ID NO: 7, a β-chain variable domain including a CDR2 including the sequence of SEQ ID NO: 8, and a β-chain variable domain including a CDR3 including the sequence of SEQ ID NO: 9.

[0039] The heterologous TCR may include, for example, an α-chain including 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 β-chain including 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 further have, for example, any of the percent identities in the α-chain and β-chain discussed herein.

[0040] Heterologous CD8 coreceptor The modified T cell includes a heterologous CD8 coreceptor. In other words, the modified T cell expresses, for example, a heterologous CD8 coreceptor on its surface.

[0041] CD8 is a cell surface glycoprotein that is naturally found on most cytotoxic T lymphocytes and mediates efficient cell-cell interactions within the immune system. CD8 acts as a coreceptor for the T cell receptor, such that CD8 and the T cell receptor together recognize antigen presented by antigen-presenting cells in the context of class I MHC molecules. The CD8 coreceptor binds to class 1 MHC and enhances TCR signaling. The functional coreceptor may be a homodimer consisting of two CD8α chains or a heterodimer consisting of one CD8α chain and one CD8β chain.

[0042] Thus, the heterologous CD8 coreceptor comprised in the modified T cell may be CD8α. In other words, the heterologous CD8 coreceptor may be a homodimer consisting of two CD8α chains. Alternatively, the heterologous CD8 coreceptor may be a heterodimer consisting of one CD8α chain and one CD8β chain. In either case, the CD8α chain may comprise, or consist of, an amino acid sequence having at least 80% (e.g., at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99% or 100%) sequence identity with SEQ ID NO: 10. Thus, the heterologous CD8 coreceptor may comprise an amino acid sequence having at least 80% (e.g., at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99% or 100%) sequence identity with SEQ ID NO: 10.

[0043] Both the CD8α chain and the CD8β chain share significant homology with the immunoglobulin variable light chain. A heterologous CD8 co-receptor may include, for example, a CD8α chain comprising (i) (1) the sequence of SEQ ID NO: 11, or (2) an amino acid sequence comprising an insertion, deletion or substitution of 1, 2 or 3 amino acids relative to the sequence of SEQ ID NO: 11 in α-chain CDR1, (ii) (1) the sequence of SEQ ID NO: 12, or (2) an amino acid sequence comprising an insertion, deletion or substitution of 1, 2 or 3 amino acids relative to the sequence of SEQ ID NO: 12 in α-chain CDR2, and / or (iii) (1) the sequence of SEQ ID NO: 13, or (2) an amino acid sequence comprising an insertion, deletion or substitution of 1, 2 or 3 amino acids relative to the sequence of SEQ ID NO: 13 in α-chain CDR3.

[0044] A heterologous CD8 co-receptor can bind to class I MHC molecules. A heterologous CD8 co-receptor can bind, for example, via the IgV-like domain of the CD8 co-receptor to, for example, the α3 portion of a class I MHC molecule. The α3 portion is typically found between residues 223 and 229 of the class I MHC molecule. The ability of a heterologous CD8 co-receptor to bind to a class I MHC molecule improves the ability of modified T cells to bind to allogeneic antigens via their heterologous TCRs. The allogeneic antigen MAGE-A4 is typically presented in a complex with a class I MHC molecule such as HLA-A * 02. A heterologous CD8 co-receptor can improve or increase the dissociation rate (k off ) of the TCR / peptide-MHCI interaction in modified cells. The improvement or increase may be compared to modified T cells that contain a heterologous TCR that binds to MAGE-A4 but lack the heterologous CD8 co-receptor. A heterologous CD8 co-receptor can assist, for example, in organizing the heterologous TCR on the surface of modified cells, thereby improving the ability of the heterologous TCR involved in the TCR / peptide-MHCI interaction. A heterologous CD8 co-receptor can bind or interact, for example, zinc-dependently with LCK (lymphocyte-specific protein tyrosine kinase), resulting in the activation of transcription factors such as NFAT, NF-κB, and AP-1.

[0045] Thus, expression of a heterologous CD8 co-receptor can confer on the engineered T cells an improved affinity and / or binding activity for MAGE-A4, and / or an improved activation upon binding to MAGE-A4. Methods for determining affinity, binding activity, and T cell activation are known in the art. Expression of a heterologous CD8 co-receptor can confer on the engineered T cells an improvement or increase in, for example, CD40L expression, cytokine production, cytotoxic activity, induction of dendritic cell maturation, or induction of dendritic cell cytokine production, in response to antigen (MAGE-A4) binding. The improvement or increase may be compared to engineered T cells that contain a heterologous TCR that binds MAGE-A4 but lack the heterologous CD8 co-receptor.

[0046] CD8α and HLA-A * Synergistic effects have been shown between CD8α and peptide antigens presented on 0201. Thus, in one aspect of the disclosure, the heterologous CD8 co-receptor may be CD8α, and the heterologous TCR may bind to the peptide antigen of MAGE-A4 that forms a complex with HLA-A * 0201. The peptide antigen may be, for example, SEQ ID NO: 1.

[0047] PD-1 axis binding antagonist Programmed cell death protein 1 (PD-1, also known as CD279) is a protein expressed on the surface of T cells that plays a role in regulating the immune response by maintaining T cell homeostasis. Ligation of PD-1 to one of its ligands (PD-L1 or PD-L2) transmits inhibitory signals within the T cell. In particular, the signal generated by PD-1 prevents phosphorylation of key TCR signaling intermediates, thereby terminating early TCR signaling and reducing T cell activation. T cell effector functions (such as proliferation, cytotoxicity, and cytokine production) are reduced, and the ability to migrate into memory T cells is impaired.

[0048] PD-L1 and PD-L2 are members of the B7 family. The PD-L1 protein is upregulated on certain activated immune cells (such as macrophages, dendritic cells, T cells, and B cells) and is also expressed on certain normal tissues. PD-L1 is also highly expressed in many cancers. PD-L2 is mainly expressed by dendritic cells and some tumors. Since many cancers express the PD-1 ligand, the PD-1 axis has established a role in cancer immune evasion and tumor resistance.

[0049] In the present disclosure, the method includes maintaining (i) a population of modified T cells and / or (ii) the function of endogenous T cells in an individual by administering a PD-1 axis-binding antagonist to the individual. Expression of the PD-1 ligand by cancers such as solid tumors makes the tumor microenvironment immunosuppressive. Thus, the function of modified T cells infiltrating the tumor can be inhibited. The endogenous antitumor T cell response can also be inhibited. In this way, the tumor can more readily evade the immune system. In the present disclosure, the PD-1 axis-binding antagonist is administered to counteract the inhibitory effect of PD-L1 and / or PD-L2 expression in the tumor microenvironment. By counteracting the inhibition, the function of modified and / or endogenous T cells can be maintained. That is, administration of the PD-1 axis-binding antagonist can sustain the function of the modified T cells included in the administration and / or their progeny. Administration of the PD-1 axis-binding antagonist can sustain the function of endogenous T cells in an individual. Preferably, administration of the PD-1 axis-binding antagonist maintains the function of the modified T cells (and / or their progeny) included in the administration, as well as the function of endogenous T cells in the individual. In either case, the endogenous T cells can be included in, for example, the tumor microenvironment. For example, the endogenous T cells may be tumor-infiltrating lymphocytes (TIL).

[0050] Maintaining the function of T cells can refer to, for example, maintaining, restoring, and / or enhancing T cell function. Maintaining T cell function can refer to, for example, maintaining T cell activation. In this way, the duration of an effective T cell response can be extended. In other words, sustained activation can be associated with an improved duration of effector functions (such as cytokine production, cytotoxicity, and / or proliferation). Sustained activation can also assist the ability of T cells to transition into memory T cells. The generation of memory T cells is advantageous as it enables the maintenance of anti-tumor immunity over a long period, for example, for several months or years. Methods for determining the activation, cytokine production, cytotoxicity, proliferation, and generation of memory T cells are known in the art.

[0051] Administration of a PD-1 axis-binding antagonist can sustain the function of engineered T cells and / or endogenous T cells by reducing exhaustion. Exhaustion can be reduced within a population of engineered T cells and / or within T cells that are progeny of a population of engineered T cells. Exhaustion can be reduced within the endogenous T cells of an individual. Exhaustion can be reduced (i) within a population of engineered T cells and / or within T cells that are progeny of a population of engineered T cells, and (ii) within the endogenous T cells of an individual. Exhausted T cells typically express high levels of PD-1 and experience loss of function. For example, exhausted T cells may have a reduced ability to produce cytokines such as IL-2 or TNFα. Exhausted T cells may have a reduced proliferative capacity. Exhausted T cells may have a reduced cytotoxic capacity. Ultimately, exhausted T cells can be targeted for destruction. Thus, exhaustion causes loss of T cell function or loss of the T cells themselves, which is disadvantageous for tumor immunity. Therefore, reducing T cell exhaustion can improve treatment outcomes.

[0052] In the context of the present disclosure, a PD-1 axis-binding antagonist is a molecule that inhibits the interaction between PD-1 and a PD-1 ligand, and / or the transmission of signals resulting from the interaction between PD-1 and a PD-1 ligand. The PD-1 ligand may be PD-L1 or PD-L2. A PD-1 axis-binding antagonist can, for example, reduce or prevent the interaction between PD-1 and a PD-1 ligand. A PD-1 axis-binding antagonist can, for example, reduce or prevent the transmission of signals resulting from the interaction between PD-1 and a PD-1 ligand. A PD-1 axis-binding antagonist can block, inhibit, or reduce the biological activity of PD-1 and / or a PD-1 ligand.

[0053] By inhibiting the interaction between PD-1 and a PD-1 ligand, and / or the transmission of signals resulting from the interaction between PD-1 and a PD-1 ligand, a PD-1 axis-binding antagonist can sustain (e.g., maintain, restore, or enhance) the function of endogenous T cells. Similarly, a PD-1 axis-binding antagonist can sustain (e.g., maintain, restore, or enhance) the function of modified T cells administered to an individual. Sustained function can be indicated, for example, by the maintenance or improvement of T cell proliferation, cytokine production, target cell killing, activation, CD28 signaling, the ability to infiltrate tumors, the ability to recognize and bind dendritic cell-presented antigens, and / or the ability to produce interferon. In this way, a PD-1 axis-binding antagonist counteracts the immunosuppressive nature of the tumor microenvironment.

[0054] The PD-1 axis-binding antagonist may be, for example, a PD-1-binding antagonist. That is, the PD-1 axis-binding antagonist may inhibit (e.g., prevent or reduce) the binding of PD-1 to its binding partner. 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 be, for example, an antibody that binds to PD-1, or an antigen-binding variant or fragment thereof. The PD-1-binding antagonist may be, for example, 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, semaprilumab, dostarlimab, JTX-4014, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, retifanlimab (INCMGA00012), AMP-224, MEDI0680 (AMP-514), sasanglimab, budigalimab, ezabenlimab (BI754091), zimberelimab (AB122). Thus, the PD-1 axis-binding antagonist may be nivolumab, pembrolizumab, semaprilumab, dostarlimab, JTX-4014, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, retifanlimab (INCMGA00012), AMP-224, MEDI0680 (AMP-514), sasanglimab, budigalimab, ezabenlimab (BI754091), zimberelimab (AB122) or any combination thereof. The PD-1 axis-binding antagonist may be, for example, nivolumab. Alternatively, the PD-1-binding antagonist may be any immunoadhesin, protein, or oligopeptide that inhibits, reduces, prevents, or interferes with signal transduction by the interaction of PD-1 with PD-L1 and / or PD-L2.

[0055] The heavy chain sequence and light chain sequence of nivolumab are shown in SEQ ID NO: 17 and SEQ ID NO: 18, respectively. The heavy chain sequence and light chain sequence of pembrolizumab are shown in SEQ ID NO: 19 and SEQ ID NO: 20, respectively. The heavy chain sequence and light chain sequence of semiprimab are shown in SEQ ID NO: 21 and SEQ ID NO: 22, respectively. The heavy chain sequence and light chain sequence of dostarlimab are shown in SEQ ID NO: 31 and SEQ ID NO: 32, respectively. One skilled in the art having the heavy chain and light chain sequences of a given antibody can identify antigen-binding variants or fragments of the antibody using methods routine in the art. An antigen-binding variant or fragment of nivolumab can include, for example, the three CDRs contained in SEQ ID NO: 17 and the three CDRs contained in SEQ ID NO: 18. An antigen-binding variant or fragment of pembrolizumab can include, for example, the three CDRs contained in SEQ ID NO: 19 and the three CDRs contained in SEQ ID NO: 20. An antigen-binding variant or fragment of semiprimab can include, for example, the three CDRs contained in SEQ ID NO: 21 and the three CDRs contained in SEQ ID NO: 22. An antigen-binding variant or fragment of dostarlimab can include, for example, the three CDRs contained in SEQ ID NO: 31 and the three CDRs contained in SEQ ID NO: 32. Methods for identifying CDRs within a heavy or light chain sequence are routine in the art.

[0056] The PD-1 axis-binding antagonist may be, for example, a PD-L1-binding antagonist. That is, the PD-1 axis-binding antagonist can inhibit (e.g., prevent or reduce) the binding to the binding partner of PD-L1. For example, the PD-1 axis-binding antagonist can inhibit the binding of PD-L1 to PD-1. The PD-L1-binding antagonist may be, for example, an antibody that binds to PD-L1, or an antigen-binding variant or fragment thereof. The PD-L1-binding antagonist may be, for example, an antibody that binds to SEQ ID NO: 15, or an antigen-binding variant or fragment thereof. Antibodies that bind to PD-L1 are known in the art and include, for example, durvalumab, atezolizumab, avelumab, BMS936559 (MDX-1105), envafolimab (KN035), and cosibelimab (CK-301). Therefore, the PD-L1 axis-binding antagonist may be durvalumab, atezolizumab, avelumab, BMS936559 (MDX-1105), envafolimab (KN035), cosibelimab (CK-301), or any combination thereof. Alternatively, the PD-L1-binding antagonist may be any immunoadhesin, protein, or oligopeptide that inhibits, reduces, prevents, or interferes with signal transduction by the interaction between PD-L1 and PD-1.

[0057] The heavy chain and light chain sequences of durvalumab are shown in SEQ ID NO: 23 and SEQ ID NO: 24, respectively. The heavy chain and light chain sequences of atezolizumab are shown in SEQ ID NO: 25 and SEQ ID NO: 26, respectively. The heavy chain and light chain sequences of avelumab are shown in SEQ ID NO: 27 and SEQ ID NO: 28, respectively. The heavy chain and light chain sequences of BMS936559 (MDX-1105) are shown in SEQ ID NO: 29 and SEQ ID NO: 30, respectively. One of ordinary skill in the art having the heavy chain and light chain sequences of a given antibody can identify antigen-binding variants or fragments of the antibody using methods routine in the art. Antigen-binding variants or fragments of durvalumab can include, for example, the three CDRs contained in SEQ ID NO: 23 and the three CDRs contained in SEQ ID NO: 24. Antigen-binding variants or fragments of atezolizumab can include, for example, the three CDRs contained in SEQ ID NO: 25 and the three CDRs contained in SEQ ID NO: 26. Antigen-binding variants or fragments of avelumab can include, for example, the three CDRs contained in SEQ ID NO: 27 and the three CDRs contained in SEQ ID NO: 28. Antigen-binding variants or fragments of BMS936559 (MDX-1105) can include, for example, the three CDRs contained in SEQ ID NO: 29 and the three CDRs contained in SEQ ID NO: 30. Methods for identifying CDRs within a heavy or light chain sequence are routine in the art.

[0058] A PD-1 axis-binding antagonist can be, for example, a PD-L2-binding antagonist. That is, a PD-1 axis-binding antagonist can inhibit (e.g., prevent or reduce) binding to the binding partner of PD-L2. A PD-L2-binding antagonist can be, for example, an antibody that binds to PD-L2, or an antigen-binding variant or fragment thereof. A PD-L2-binding antagonist can be, for example, an antibody that binds to SEQ ID NO: 16, or an antigen-binding variant or fragment thereof. Alternatively, a PD-L2-binding antagonist can be any immunoadhesin, protein, or oligopeptide that inhibits, reduces, prevents, or interferes with signal transduction by the interaction of PD-L2 and PD-1.

[0059] When the PD-1 axis-binding antagonist is an antibody (such as a known antibody or its antigen-binding variant), the PD-1 axis-binding antagonist may be, for example, a monoclonal antibody, a human or humanized antibody, a full-length antibody, a diabody, a linear antibody, or a single-chain antibody molecule. The antibody isotype can be selected from any of the five classes of immunoglobulins having heavy chains called alpha, delta, epsilon, gamma, and mu (M), namely IgA, IgD, IgE, IgG, and IgM. The gamma and alpha class antibodies may be any of the subclasses IgG1, IgG2A, IgG2B, IgG3, IgG4, IgA1, 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, for example, Fv, Fab, Fab’, Fab’-SH, F(ab’)2, or scFv.

[0060] When the PD-1 axis-binding antagonist is an immunoadhesin, the immunoadhesin may include an adhesion domain and an immunoglobulin constant domain that confer binding activity to a PD-1 axis component (such as PD-1, PD-L1, or PD-L2). The immunoglobulin constant domain may be derived from any isotype such as IgG1, IgG2, IgG2A, IgG2B, IgG3, IgG4 subtype, IgA, IgA1, IgA2, IgE, IgD, or IgM. The immunoglobulin constant domain may include, for example, (i) the hinge, CH2, and CH3, or (ii) the hinge, CH1, CH2, and CH3 regions of the immunoglobulin molecule. Thus, the immunoadhesin may include (a) the extracellular or PD-1-binding portion of PD-L1 or PD-L2, or the extracellular or PD-L1 or PD-L2-binding portion of PD-1, fused to (b) the constant domain of the immunoglobulin sequence.

[0061] Administration In the present disclosure, the method comprises maintaining the function of (i) a population of modified T cells and / or (ii) endogenous T cells in an individual by: (a) administering to the individual a population of modified T cells; and (b) administering a first dose of a PD-1 axis-binding antagonist.

[0062] The population can contain any number of modified T cells that is therapeutically effective. The number of modified T cells for a given individual can depend on factors such as the cancer being treated, the severity or stage of the cancer, and the age of the patient. The number administered can thus depend on the judgment of the physician and can be specific to each subject. By way of example, the population can contain from about 0.8×10 9 to about 10×10 9 modified T cells, e.g., from about 0.8×10 9 to about 1.2×10 9 modified T cells, from about 1.2×10 9 to about 6×10 9 modified T cells, or from about 1.0×10 9 to 10×10 9 modified T cells. The population can contain, for example, 1.0×10 9 modified T cells, about 5.0×10 9 modified T cells, or about 10×10 9 modified T cells.

[0063] Typically, the population of modified T cells is administered as a single dose. However, additional doses (e.g., 2 or more, 3 or more, 4 or more, or 5 or more) may be administered depending on patient factors and the judgment of the physician.

[0064] Typically, the population of modified T cells is administered intravenously. However, any suitable route, such as intramuscular, subcutaneous, intradermal, transdermal, or intraperitoneal, may be used.

[0065] The first dose of the PD-1 axis-binding antagonist is administered about 3 to about 5 weeks after the administration of the population of modified T cells. The first dose of the PD-1 axis-binding antagonist can be administered, for example, about 3 weeks, about 4 weeks, or about 5 weeks after the administration of the population of modified T cells. The first dose of the PD-1 axis-binding antagonist can be administered, for example, about 21 to about 35 days, for example about 25 to about 30 days, after the administration of the population of modified T cells. The first dose of the PD-1 axis-binding antagonist can be administered, for example, about 21 days, about 22 days, about 23 days, about 24 days, about 25 days, about 26 days, about 27 days, about 28 days, about 29 days, about 30 days, about 31 days, about 32 days, about 33 days, about 34 days, or about 35 days after the administration of the population of modified T cells. Preferably, the first dose of the PD-1 axis-binding antagonist is administered about 4 weeks (i.e., about 28 days) after the administration of the population of modified T cells. For example, the first dose of the PD-1 axis-binding antagonist can be administered about 25 to about 30 days (such as about 25 days, about 26 days, about 27 days, about 28 days, about 29 days, or about 30 days) after the administration of the population of modified T cells.

[0066] The inventors have identified that this window between the administration of a population of modified T cells and the administration of the first dose of a PD-1 axis-binding antagonist is surprisingly advantageous. As demonstrated in the examples, the inventors have demonstrated that the modified T cells peak at about 13-16 days post-administration and persist at optimal levels for over 24 weeks. In responsive individuals (determined by Best Overall Response, BOR), this unexpectedly early peak in T cell persistence led to a reduction in the diameter of target lesions at the earliest study time point, 4 weeks after administration. This indicates that the modified T cells have a therapeutic effect at a surprisingly early time point. Furthermore, the examples demonstrate that TILs are present in the target tumor by 4 weeks after administration of the modified T cells, and that the TILs contain both modified T cells and T cells endogenous to the individual. Thus, the inventors have shown that a PD-PD-L1-PD-L2 axis inhibitor can be optimally administered 4 weeks after or around the time of infusion of the modified T cells. The data indicate that at this time, the modified endogenous T cells are infiltrating the tumor and initiating an effective anti-tumor response. Administration of a PD-PD-L1-PD-L2 axis inhibitor 4 weeks after or around the time of infusion coincides with the time when T cell activity / functionality is strongly manifested and thus serves to sustain T cell activity / functionality by delaying T cell exhaustion mediated by PD-PD-L1-PD-L2 axis signaling. In this way, effector function can be extended and the durability of activated T cells increased.

[0067] Aligning the timing of administration of the first dose of a PD-1 axis-binding antagonist to coincide with the presence of TILs and / or the onset of the therapeutic effect of the modified T cells is beneficial in order to maintain the activation (and thus function) of adoptively transferred T cells and endogenous T cells at therapeutically effective levels. It also promotes the generation of memory T cells, thereby conferring long-term anti-tumor immunity and thus improving the patient's outcome.

[0068] The method may include administering one or more additional doses of a PD-1 axis-binding antagonist, for example. The purpose of the one or more additional administrations of the PD-1 axis-binding antagonist may be to maintain the effect achieved by the administration of the initial dose. Each of the one or more additional doses may include the same PD-1 axis-binding antagonist as the initial dose, or a PD-1 axis-binding antagonist different from the initial dose.

[0069] One or more additional doses of the PD-1 axis-binding antagonist may be administered at any suitable interval. Suitable dosing intervals for the PD-1 axis-binding antagonist are known in the art. The one or more additional doses may be administered, for example, starting 2 weeks after the administration of the initial dose, once every about 2 weeks (Q2W). The one or more additional administrations may be administered, for example, starting 3 weeks after the administration of the initial dose, once every about 3 weeks (Q3W). The one or more additional doses may be administered, for example, starting 4 weeks after the administration of the initial dose, once every about 4 weeks (Q4W). The one or more additional administrations may be administered, for example, starting 5 weeks after the administration of the initial dose, once every about 5 weeks (Q5W). The one or more additional doses may be administered, for example, starting 6 weeks after the administration of the initial dose, once every about 6 weeks (Q6W). In a preferred embodiment of the present disclosure, the one or more additional doses are administered starting 4 weeks after the administration of the initial dose, once every about 4 weeks (Q4W). Any number of additional doses 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, fifteen or more, twenty or more, twenty-five or more, thirty or more, thirty-five or more, forty or more, forty-five or more, or fifty or more additional doses. Additional doses can be administered until disease progression, unacceptable toxicity, withdrawal of consent, or death.

[0070] The initial dose of the PD-1 axis-binding antagonist, and / or any additional dose of the PD-1 axis-binding antagonist, may comprise any therapeutically effective amount of the PD-1 axis-binding antagonist. The amount for a given individual may depend on factors such as the cancer being treated, the severity or stage of the cancer, and the age of the patient. The number of administrations may thus depend on the judgment of the physician and may be specific to each subject. By way of example, the initial dose of the PD-1 axis-binding antagonist, and / or any additional dose of the PD-1 axis-binding antagonist, may comprise from about 200 mg to about 700 mg of the PD-1 axis-binding antagonist, such as from about 300 mg to about 600 mg, from about 400 mg to about 500 mg, or from about 450 mg to 500 mg of the PD-1 axis-binding antagonist. For example, the initial dose of the PD-1 axis-binding antagonist, and / or any additional dose of the PD-1 axis-binding antagonist, may comprise about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, or about 700 mg of the PD-1 axis-binding antagonist. The initial dose of the PD-1 axis-binding antagonist, and / or any additional dose of the PD-1 axis-binding antagonist, may comprise, for example, from about 450 mg to about 500 mg of the PD-1 axis-binding antagonist, such as about 480 mg of the PD-1 axis-binding antagonist. In a preferred embodiment of the present disclosure, the initial dose of the PD-1 axis-binding antagonist, and any additional dose of the PD-1 axis-binding antagonist, comprise 480 mg of nivolumab.

[0071] By way of example, the PD-1 axis-binding antagonist may be administered at an initial dose of about 480 mg, for example, about 4 weeks after administration of the population of modified T cells. One or more additional doses may be administered starting about 4 weeks after administration of the initial dose and at about 4-week intervals. Each additional dose may comprise about 480 mg of the PD-1 axis-binding antagonist. The PD-1 axis-binding antagonist may be, for example, nivolumab.

[0072] In another embodiment, the PD-1 axis-binding antagonist can be administered at an initial dose of about 240 mg, for example, about 4 weeks after administration of the population of modified T cells. One or more additional doses can be administered about every 2 weeks, starting 2 weeks after administration of the initial dose. Each additional dose can contain about 240 mg of the PD-1 axis-binding antagonist. The PD-1 axis-binding antagonist can be, for example, nivolumab.

[0073] Typically, the PD-1 axis-binding antagonist is administered intravenously. However, any suitable route, such as intramuscular, subcutaneous, intradermal, transdermal, or intraperitoneal routes, may be used.

[0074] In some aspects of the present disclosure, the method includes administering lymphodepleting chemotherapy to an individual prior to administration of a population of modified T cells. That is, lymphodepleting chemotherapy can be administered prior to step (a). Lymphodepleting chemotherapy can be administered, for example, from about 14 days prior to step (a) to about 1 day prior to step (a), such as from about 13 days prior to step (a) to about 2 days prior to step (a), from about 12 days prior to step (a) to about 3 days prior to step (a), from about 11 days prior to step (a) to about 4 days prior to step (a), from about 10 days prior to step (a) to about 5 days prior to step (a), from about 9 days prior to step (a) to about 6 days prior to step (a), from about 8 days prior to step (a) to about 7 days prior to step (a), from about 10 days prior to step (a) to about 1 day prior to step (a), from about 9 days prior to step (a) to about 2 days prior to step (a), from about 8 days prior to step (a) to about 3 days prior to step (a), from about 7 days prior to step (a) to about 4 days prior to step (a), or from about 6 days prior to step (a) to about 5 days prior to step (a). Preferably, lymphodepleting chemotherapy is administered from about 7 days prior to step (a) to about 4 days prior to step (a). The purpose of lymphodepleting chemotherapy may be to deplete the lymphocyte compartment of the individual so as to provide a space in which the adoptively transferred modified T cells can proliferate. In this way, the effect of a given dose of modified T cells can be maximized. Lymphodepleting chemotherapy can include any suitable lymphotoxic agent. Lymphotoxic agents and suitable dosages are known in the art. Lymphodepleting chemotherapy can include, for example, fludarabine and / or cyclophosphamide. Typically, lymphodepleting chemotherapy is administered intravenously. However, any suitable route, such as intramuscular, subcutaneous, intradermal, transdermal, or intraperitoneal routes, may be used.

Example

[0075] Dosing schedule of ADP-A2M4CD8 with a PD-1 axis-binding antagonist Introduction ADP-A2M4CD8 specific peptide enhanced affinity receptor (SPEAR™) T cells are genetically engineered to target the tumor antigen MAGE-A4 in the context of appropriate human leukocyte antigen (HLA) expression. ADP-A2M4CD8 are autologous CD4 and CD8 positive T cells transduced with a self-inactivating (SIN) lentiviral vector expressing a high-affinity MAGE-A4 specific T cell receptor (TCR) and an additional CD8α co-receptor.

[0076] The affinity-optimized TCR (ADP-A2M4 TCR) comprises an α-chain variable domain having the amino acid sequence of SEQ ID NO: 2 and a β-chain variable domain having the amino acid sequence of SEQ ID NO: 3. When expressed in T cells, the signal peptide is cleaved from SEQ ID NOs: 2 and 3 prior to surface expression. The A2M4 TCR targets the tumor antigen MAGE-A4 and activates engineered T cells. This recognizes the MAGE-A4230-239 (GVYDGREHTV; SEQ ID NO: 1) peptide sequence derived from MAGE-A4 when present in the HLA-A * 02-GVYDGREHTV antigen complex.

[0077] The CD8α co-receptor contained in ADP-A2M4CD8 SPEAR™ T cells is designed to provide additional functionality to CD4 T cells. Since CD4+ T cells have weak effector functions in response to class I antigens, the CD8α co-receptor was introduced together with the TCR to increase TCR binding activity and enhance the multifunctional response of engineered CD4+ T cells against MAGE-A4 positive tumors. Co-expression of CD8α confers the ability of CD8+ killer T cells to CD4+ helper T cells while maintaining / enhancing the helper cell ability of CD4+ T cells. Addition of the CD8α co-receptor directly affects TCR binding to HLA-peptide complexes in CD4+ T cells and enhances CD4+ T cell effector functions. Thus, ADP-A2M4CD8 SPEAR™ T cells are designed to improve ADP-A2M4-expressing T cells.

[0078] This has been confirmed in preclinical in vitro assays, and ADP-A2M4CD8 showed a clear improvement in T cell activation, as measured by an increase in CD40L surface expression, particularly in the CD4+ fraction, compared to ADP-A2M4-expressing T cells (when cultured with antigen-positive cells). When dendritic cells (DCs) were included in the co-culture, a marked improvement was seen with ADP-A2M4CD8 T cells. Cytokine release from both DCs (IL-12, MIG) and T cells (IFNγ, IL-2, and other Th1) was improved compared to cultures containing ADP-A2M4 cells. Furthermore, when ADP-A2M4CD8 was transduced, it was found that CD4+ T cells changed to a state with an effective cytotoxic function, as MAGE-A4-positive 3D microspheres could not be killed. Therefore, CD4+ T cells transduced with ADP-A2M4CD8 exhibit not only CD4+ helper function but also improved T cell effector function.

[0079] Despite this improvement, preliminary results from one trial showed that 55% of patients did not respond to ADP-A2M4CD8 treatment. Therefore, a plan to improve the response to ADP-A2M4CD8 therapy is desired. This example relates to the improvement of the response to ADP-A2M4CD8 therapy using agents that help maintain an activated T cell response.

[0080] In this regard, binding antagonists of the PD-L1 / PD-1 axis are considered. PD-1 is expressed on T cells, and PD-L1 is expressed on various different cancer cells. The physiological role of PD-1 is to maintain T cell homeostasis by restricting T cell activation and proliferation. Ligation of PD-L1 to PD-1 transmits an inhibitory exhaustion signal that reduces cytokine production and inhibits T cell proliferation. In particular, when T cells experience simultaneous engagement of the TCR and PD-1, the signal generated by PD-1 prevents phosphorylation of important TCR signaling intermediates, thereby terminating early TCR signaling and reducing T cell activation. T cell effector functions and the transition to memory T cells are impaired. In this way, the adaptive anti-cancer immune response is restricted and tumor tolerance is promoted.

[0081] PD-1 pathway blockade is hypothesized to improve the response to ADP-A2M4CD8 therapy.

[0082] Materials and Methods Subjects were selected to investigate the effect of PD-1 pathway blockade on the response to ADP-A2M4CD8 therapy. Briefly, subjects with a histologically or cytogenetically confirmed diagnosis of urothelial cancer, esophageal, esophagogastric junction cancer, gastric cancer, non-small cell lung cancer (NSCLC), head and neck cancer, ovarian cancer, melanoma, or endometrial cancer were pre-screened to determine the appropriate human leukocyte antigen (HLA) and tumor antigen status. For inclusion in the study, subjects had to be 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 domain. Patients positive for HLA-A * 02:05 were excluded from the study because alloreactivity from ADP-A2M4 and ADP-A2M4CD8 was seen against two HLA-A * 02:05 positive cell lines. HLA-A * 02:07 or any A* Patients having any of the 02 null alleles as the only HLA-A * 02 alleles were also excluded due to reduced activity of these alleles. The subject must also not have a tumor showing MAGE-A4 expression defined as ≥ 30% of tumor cells being ≥ 2+ as determined by immunohistochemistry (IHC).

[0083] For the treatment and manufacture of ADP-A2M4CD8, autologous cells were collected by leukapheresis from registered subjects. The heterologous TCRs contained in ADP-A2M4CD8 T cells include the α-chain sequence set forth in SEQ ID NO: 2 and the β-chain sequence set forth in SEQ ID NO: 3. The heterologous CD8 co-receptors contained in ADP-A2M4CD8 T cells include two CD8α chains each containing the amino acid sequence of SEQ ID NO: 10. The surface-expressed heterologous TCR and the surface-expressed heterologous CD8 co-receptor do not include a signal sequence.

[0084] A baseline tumor evaluation was obtained prior to treatment. The subject was then administered lymphodepleting chemotherapy using fludarabine and cyclophosphamide (-7 days to -4 days). The subject received a single intravenous infusion of ADP-A2M4CD8 on day 1. Approximately 60 subjects were treated in this manner (Groups 1 to 3 in Table 1 and Figure 1 below). An additional subject group, Group 4, was further treated with a PD-1 axis antagonist (nivolumab, and Figure 2).

[0085]

Table 1

[0086] The initial dose selected for ADP-A2M4CD8 was 1×10 9 transduced cells (range: 0.8×10 9 ~1.2×10 9 transduced cells). To date, doses of ADP-A2M4 cells in the range of 0.1×10 9 ~10×10 9 have been used clinically. The peak expansion of ADP-A2M4 cells after infusion is low, 1×109 At sub-therapeutic doses, persistence was transient and there was no potential for biological activity. The target antigen for both ADP-A2M4 and ADP-A2M4CD8 is MAGE-A4, but the effect of CD8α modification in humans was not known. Therefore, to maintain a positive benefit:risk balance, the starting dose in the study was selected to be potentially effective but below the tolerated dose of ADP-A2M4. The starting dose of ADP-A2M4CD8 is further supported by the experience of NY-ESO-1c259T in patients with synovial sarcoma. Subjects were injected with a transduced cell dose of NY-ESO-1c259T with a median of 3.6×10 9 (range, 0.45×10 9 ~14.4×10 9 )(Araujo, 2019). Two subjects administered a dose of less than 1×10 9 showed no response and had progressive disease (PD) by 12 weeks post-injection, indicating that response is likely to be observed at a dose of transduced T cells of 1×10 9 or greater.

[0087] Subjects were monitored immediately after injection (days 1 - 8). Subjects were monitored weekly until 4 weeks post-injection. Subjects were then monitored at 6, 8, 12, 16, and 24 weeks and thereafter at least every 3 months until disease progression. Tumor response was evaluated according to Response Evaluation Criteria in Solid Tumors (RECIST) v1.1. Additional data collected included the following. - Core needle biopsies to directly assess the "immune status" within the tumor at baseline and during the course of the study. - Cytokine levels in serum at baseline and during the course of the study. - Humoral immune responses to tumor antigens in serum at baseline and during the course of the study. - Antibodies to ADP-A2M4CD8 in serum at baseline and during the course of the study. - Soluble markers representing tumors and their microenvironment using liquid biopsy. For example, markers of circulating tumor cells (CTC), exosomes, and cell-free DNA (cfDNA) produced by dying tumor cells can be used to monitor both the molecular signature of tumor burden (including the expression of target antigens) and the immune response. Analysis of such soluble markers enables the estimation of overall tumor burden and gene profiling, including the expression and mutation profiling of MAGE-A4 mRNA. Additionally, analysis of such soluble markers enables the systemic evaluation of the immune response. - Phenotype and activity before and after gene-modified T cell injection. Related assays can be performed using blood and, if resection is performed, the tumor. Assays include (i) phenotypic analysis for determination of the T cell lineage in cell products after injection and in blood (and, if resection is performed, the tumor), (ii) quantification of the senescence and activation status of immune subsets from PBMC, (iii) analysis of gene expression or epigenetic profiles to reflect the phenotypic and functional state of the cells, and / or (iv) direct functional evaluation of the cells. - Persistence of the injected modified cells and correlation with the therapeutic effect. Persistence can be determined by data on the copy of gene-modified DNA per 1 μg of DNA and / or the number of transduced cells or the total lymphocyte count per 1 μL. Established methods include (i) quantification of ADP-A2M4CD8 cells by quantitative PCR of the transgene from DNA extracted from cryopreserved PBMC, and (ii) quantification and phenotyping of ADP-A2M4CD8 cells by flow cytometry, DNA, and RNA analysis from cryopreserved PBMC.

[0088] Results Throughout the study, up to 10×10 9ADP-A2M4CD8 was administered at a dose and showed good tolerance. The treatment-emergent adverse events (TEAEs) were consistent with the underlying diseases and known adverse event profiles using non-myeloablative lymphocyte depletion or cancer immunotherapy. No dose-limiting toxicity was observed. Ten subjects reported serious adverse events considered to be related to ADP-A2M4CD8. Most of the notable adverse events were consistent with those typically experienced by cancer subjects receiving cytotoxic chemotherapy or cancer immunotherapy.

[0089] As of May 24, 2021, 20 subjects (1 with MRCLS, 5 with EGJ cancer, 6 with ovarian cancer, 2 with head and neck squamous cell carcinoma, 2 with esophageal cancer, 1 with melanoma, 1 with NSCLC, 1 with synovial sarcoma, and 1 with urothelial cancer) had been administered ADP-A2M4CD8 (in the range of 1.0 to 590 million transduced cells), and treatment benefit / tumor regression was observed in patients with EGJ cancer, head and neck cancer, esophageal cancer, and ovarian cancer. On this day, overall, across all dose groups and tumor types, the best overall response (BOR) was 1 subject (5%) with a confirmed complete response, 4 subjects (20%) with a confirmed partial response (cPR), 2 subjects (10%) with an unconfirmed partial response (uPR), 8 subjects (40%) with stable disease (SD), 3 subjects (15%) with progressive disease (PD), and 1 subject (5%) lacking this evaluation. One subject with EGJ cancer had a cPR by RECIST and, as of the cutoff date of May 24, 2021, had a progression-free survival (PFS) of over 36 weeks. Two head and neck cancer patients had a cPR. Tumor regression (the sum of the diameters of the target lesions reaching the nadir) of 16.3% reduction from baseline was observed in ovarian cancer patients.

[0090] Studies of SPEAR™ T cells indicate that PD-1 is expressed on CD8+ T cells and CD4+ T cells in the majority of apheresis products, as well as in both transduced and untransduced T cells in the manufactured products. In addition, induction of intratumoral PD-L1 was observed early after T cell infusion in one subject with synovial sarcoma and another subject with ovarian cancer, and was associated with an increase in transduced and untransduced SPEAR™ T cells. Furthermore, in vitro experiments showed that PD-1 blockade by pembrolizumab partially restored IFN-γ production by pre-activated A2M4 SPEAR™ T cells during restimulation in multiple donors. PD-1 / PD-L1 blockade has also been demonstrated to enhance the effects of CAR-T therapy in vivo and ex vivo in hematologic malignancies and solid tumors.

[0091] Therefore, PD-1 pathway blockade may maintain the function of infused ADP-A2M4 CD8 T cells, such as effector function and the ability to migrate into memory T cells. Generation of memory T cells is advantageous in the treatment of tumors because the presence of memory T cells in the subject may allow for a long-term (e.g., months to years) anti-tumor effect that continues to suppress the tumor even when ADP-A2M4 CD8 T cell therapy is discontinued. Furthermore, the inventors propose that PD-1 pathway blockade may improve the patient response to ADP-A2M4 CD8 T cell therapy by engaging the patient's own immune system. In particular, PD-1 axis antagonists may bind to endogenous T cells, such as those that infiltrate the tumor microenvironment. PD-1 pathway blockade may maintain the function of these endogenous T cells, such as effector function and the ability to migrate into memory T cells. Therefore, PD-1 pathway blockade may (i) sustain the function of ADP-A2M4 CD8 T cells and / or (ii) maintain the function of endogenous T cells such that they can contribute to an anti-tumor response. In these ways, PD-1 pathway blockade may enhance and / or extend the effect of a single dose of ADP-A2M4 CD8 T cells administered to a subject.

[0092] In this regard, this study enabled the identification of an optimal dosing schedule for administering a PD-1 axis inhibitor in combination with ADP-A2M4 CD8 T cells. Tables 2 and 3 below summarize the peak persistence of ADP-A2M4 CD8 T cells and the time to peak persistence in subjects administered ADP-A2M4 CD8 T cells by infusion. Figure 4 is taken from the prior art and provides comparative data on the persistence of a single infusion of CAR T cells. Figure 5 shows the effect of a single infusion of ADP-A2M4 CD8 T cells on the target lesion (tumor).

[0093]

Table 2

[0094] The data show that ADP-A2M4 CD8 T cells optimally persist over a long period exceeding 24 weeks (see Tables 2 and 3). This is in contrast to CAR T cells, which have been demonstrated in the prior art to only persist at low levels after an initial peak at 7-14 days post-infusion (Figure 4).

[0095] The data further demonstrate that tumors show an early response to ADP-A2M4 CD8 T cells. The first tumor evaluation was performed at week 4. Figure 5 shows that even at this early time point, ADP-A2M4 CD8 T cells induce anti-tumor activity, which results in a reduction in tumor burden and, in some cases, meets the RECIST criteria for partial response.

[0096] The inventors also found that intratumoral ADP-A2M4 CD8 T cells were detected in 75% of evaluable biopsies obtained from patients administered with ADP-A2M4 CD8 T cells. Figure 6A shows that the tumor contains other T cells in addition to A2M4 CD8 T cells. That is, the tumor contains endogenous T cells derived from the patient, as well as the administered therapeutic T cells. As shown in Figure 6B, administration of ADP-A2M4 CD8 T cells can increase the infiltration of the patient's own T cells into the tumor. In particular, the box plot in Figure 6B shows the changes in the number of malignant cells, CD4 helper T cells, cytotoxic T cells, and regulatory T cells from baseline to after ADP-A2M4 CD8 T cell injection. The phenotype is also considered. In patients with urothelial cancer having a stable state (SD) after injection, the native (PDL1- and PDL1-Ki67-) type of malignant cells decreased from baseline to after injection. PDL1+ and proliferative PDL1+ malignant cells decreased more slowly from baseline to after injection, consistent with their having a stronger immunosuppressive phenotype and a reduced sensitivity to killing by immune cells. The native types of CD4 T helper cells, cytotoxic T cells, and regulatory T cells increased after injection, and PDL1+ CD4 T cells and PDL1+ cytotoxic T cells showed a slightly higher rate of increase. In esophageal cancer patients with progressive disease (PD), the native (PDL1- and PDL1-Ki67-) type of malignant cells increased very slightly from baseline to after injection. The density change rate of PDL1+ and proliferative PDL1+ malignant cells was increased compared to native malignant cells, consistent with their having a stronger immunosuppressive phenotype and a reduced sensitivity to killing by immune cells. The native types of CD4 T helper cells and especially cytotoxic T cells decreased after baseline, as predicted for progressive disease. Regulatory T cells showed a rapid increase from baseline to after injection. PDL1+ cytotoxic T cells showed a slightly slower rate of decrease from baseline to after injection, and PDL1+ proliferative cytotoxic T cells even showed an increase in density. PDL1+ and proliferative PDL1+ CD4 T cells showed an increase in density from baseline to after injection compared to the native type.Shown in Table 2 and FIGS. 3-6, the data described above demonstrate that modified endogenous TILs are present up to 4 weeks after administration of ADP-A2M4CD8 T cells and that tumor size reduction occurs concomitantly. Thus, the data indicate that a PD-PD-L1-PD-L2 axis inhibitor can be optimally administered 4 weeks or around that time after injection of ADP-A2M4CD8 T cells. In this regard, many patients in the initial clinical trials using either an anti-PD1 or anti-PD-L1 antibody showed an objective clinical response. The mechanism of such response is thought to be the sustained T cell activity / functionality of both engineered and naturally occurring T cells by delaying T cell exhaustion mediated by PD-PD-L1-PD-L2 axis signaling. Thus, administration of a PD-PD-L1-PD-L2 axis inhibitor at 4 weeks or around that time is thought to extend effector function and ultimately increase the durability of activated T cells.

[0097] Treatment with a PD-PD-L1-PD-L2 axis inhibitor at or around the fourth week also has a favorable meaning in terms of safety and tolerability. New data show that after four weeks, most patients recover from any safety events associated with either lymphopenia (cytopenia) or ADP-A2M4CD8 T cell infusion (CRS). Thus, administration of a PD-PD-L1-PD-L2 axis inhibitor at or around the fourth week also prevents overlapping toxicities and is considered to be well tolerated by patients. It is surprising that a PD-PD-L1-PD-L2 axis inhibitor can be effectively administered four weeks or around four weeks after administration of ADP-A2M4CD8 T cells. Prior art regarding administration of T cells with a PD-PD-L1-PD-L2 axis inhibitor (e.g., Cao et al. (2019), Anti-CD19 Chimeric Antigen Receptor T Cells in Combination With Nivolumab Are Safe and Effective Against Relapsed / Refractory B-Cell Non-hodgkin Lymphoma, Frontiers in Oncology, 9:767, doi: 10.3389 / fonc.2019.00767) shows that the administered T cells reach a peak around 7 to 14 days and then decrease to very low levels. Thus, the prior art teaches that a PD-PD-L1-PD-L2 axis inhibitor should be administered immediately after therapeutic T cells, e.g., on day 3.

[0098] SEQ ID NO: 1 - MAGE-A4 230-239 GVYDGREHTV SEQ ID NO: 2 - MAGE-A4 TCR α-chain (CDRs are underlined in bold, signal sequence is underlined in italics).

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Claims

**Claim 1** A method of treating cancer in an individual, comprising: (a) administering to the individual a population of modified T cells comprising a heterologous CD8 coreceptor and a heterologous TCR capable of binding to MAGE-A4; and (b) maintaining in the individual the function of (i) the population of modified T cells and / or (ii) endogenous T cells by administering to the individual a first dose of a PD-1 axis-binding antagonist about 3 to about 5 weeks after step (a). **Claim 2** A population of modified T cells for use in a method of treating cancer in an individual, wherein the modified T cells comprise a heterologous CD8 coreceptor and a heterologous TCR capable of binding to MAGE-A4, and the method comprises: (a) administering the population to the individual; and (b) maintaining in the individual the function of (i) the population of modified T cells and / or (ii) endogenous T cells by administering to the individual a first dose of a PD-1 axis-binding antagonist about 3 to about 5 weeks after step (a). **Claim 3** A PD-1 axis-binding antagonist for use in a method of treating cancer in an individual, wherein the method comprises: (a) administering to the individual a population of modified T cells comprising a heterologous CD8 coreceptor and a heterologous TCR capable of binding to MAGE-A4; and (b) maintaining in the individual the function of (i) the population of modified T cells and / or (ii) endogenous T cells by administering to the individual a first dose of the PD-1 axis-binding antagonist about 3 to about 5 weeks after step (a). **Claim 4** The method according to claim 1, the population for use according to claim 2, or the PD-1 axis-binding antagonist for use according to claim 3, wherein the first dose of the PD-1 axis-binding antagonist is administered to the individual about 4 weeks after administration of the population of modified T cells. **Claim 5** The method further comprises administering one or more additional doses of the PD-1 axis-binding antagonist, optionally, wherein the one or more additional doses are administered once every four weeks (Q4W) starting four weeks after the administration of the initial dose, the method according to claim 1 or 4, the population for use according to claim 2 or 4, or the PD-1 axis-binding antagonist for use according to claim 3 or 4.

6. The administration of the PD-1 axis-binding antagonist reduces exhaustion in (i) within the population of the modified T cells and / or in T cells that are descendants of the population of the modified T cells, and / or (ii) in endogenous T cells in the individual, the method according to any one of claims 1, 4, and 5, the population for use according to any one of claims 2, 4, and 5, or the PD-1 axis-binding antagonist for use according to any one of claims 3 to 5.

7. The heterologous TCR binds to SEQ ID NO: 1, the method according to any one of claims 1 and 4 to 6, the population for use according to any one of claims 2 and 4 to 6, or the PD-1 axis-binding antagonist for use according to any one of claims 3 to 6.

8. The heterologous TCR comprises an α-chain amino acid sequence having at least 80% sequence identity with SEQ ID NO: 2, the method according to any one of claims 1 and 4 to 7, the population for use according to any one of claims 2 and 4 to 7, or the PD-1 axis-binding antagonist for use according to any one of claims 3 to 7.

9. The heterologous TCR comprises a β-chain amino acid sequence having at least 80% sequence identity with SEQ ID NO: 3, the method according to any one of claims 1 and 4 to 8, the population for use according to any one of claims 2 and 4 to 8, or the PD-1 axis-binding antagonist for use according to any one of claims 3 to 8.

10. The CD8 co-receptor is CD8α, the method according to any one of claims 1 and 4 to 9, the population for use according to any one of claims 2 and 4 to 9, or the PD-1 axis-binding antagonist for use according to any one of claims 3 to 9.

11. The method according to any one of claims 1 and 4 to 10, the population for use according to any one of claims 2 and 4 to 10, or the PD-1 axis-binding antagonist for use according to any one of claims 3 to 10, wherein the CD8 co-receptor comprises an amino acid sequence having at least 80% sequence identity with SEQ ID NO:

10.

12. The method according to any one of claims 1 and 4 to 11, the population for use according to any one of claims 2 and 4 to 11, or the PD-1 axis-binding antagonist for use according to any one of claims 3 to 11, wherein the modified T cell is autologous to the individual.

13. The method comprises (i) obtaining peripheral blood mononuclear cells (PBMCs) from the individual, (ii) selecting T cells from the PBMCs, and (iii) producing a population by modifying the selected T cells to express a heterologous CD8 co-receptor and a heterologous TCR capable of binding to MAGE-A4, the method, population for use, or PD-1 axis-binding antagonist for use according to claim 12.

14. The method according to any one of claims 1 and 4 to 13, the population for use according to any one of claims 2 and 4 to 13, or the PD-1 axis-binding antagonist for use according to any one of claims 3 to 13, wherein the method comprises administering lymphodepleting chemotherapy to the individual prior to administration of the population of modified T cells.

15. The group comprises 0.8×10 ９ to 10×10 ９ modified T cells, and optionally, the group comprises 0.8×10 ９ to 1.2×10 ９ modified T cells, 1.2×10 ９ to 6×10 ９ modified T cells, or 1.0×10 ９ to 10×10 ９ modified T cells, a method according to any one of claims 1 and 4 to 14, a group for use according to any one of claims 2 and 4 to 14, or a PD-1 axis binding antagonist for use according to any one of claims 3 to 14.

16. The population is about 1.0×10 ９ modified T cells, about 5.0×10 ９ modified T cells, or about 10×10 ９ modified T cells, a method according to any one of claims 1 and 4 to 15, a population for use according to any one of claims 2 and 4 to 15, or a PD-1 axis binding antagonist for use according to any one of claims 3 to 15.

17. The method according to any one of claims 1 and 4 to 16, the population for use according to any one of claims 2 and 4 to 16, or the PD-1 axis-binding antagonist for use according to any one of claims 3 to 16, wherein the population of modified T cells is administered as a single dose.

18. The method according to any one of claims 1 and 4 to 17, the population for use according to any one of claims 2 and 4 to 17, or the PD-1 axis-binding antagonist for use according to any one of claims 3 to 17, wherein the population of modified T cells is administered intravenously.

19. The method according to any one of claims 1 and 4 to 18, the population for use according to any one of claims 2 and 4 to 18, or the PD-1 axis-binding antagonist for use according to any one of claims 3 to 18, wherein the PD-1 axis-binding antagonist is a PD1-binding antagonist.

20. The method according to claim 19, the population for use, or the PD-1 axis-binding antagonist for use, wherein the PD1-binding antagonist is an antibody that binds to PD-1.

21. The method according to claim 20, the population for use, or the PD-1 axis-binding antagonist for use, wherein the antibody is nivolumab.

22. The method according to any one of claims 1 and 4 to 21, the population for use according to any one of claims 2 and 4 to 21, or the PD-1 axis-binding antagonist for use according to any one of claims 3 to 21, wherein each dose of the PD-1 axis-binding antagonist comprises 200 mg to 700 mg of the PD-1 axis-binding antagonist.

23. The method according to claim 22, the population for use, or the PD-1 axis-binding antagonist for use, wherein each dose of the PD-1 axis-binding antagonist comprises 300 mg to 600 mg, 400 mg to 500 mg, 450 mg to 500 mg, or 480 mg of the PD-1 axis-binding antagonist.

24. The method according to any one of claims 1 and 4 to 23, the population for use according to any one of claims 2 and 4 to 23, or the PD-1 axis-binding antagonist for use according to any one of claims 3 to 23, wherein each dose of the PD-1 axis-binding antagonist comprises 480 mg of nivolumab.

25. The method according to any one of claims 1 and 4 to 24, the population for use according to any one of claims 2 and 4 to 24, or the PD-1 axis-binding antagonist for use according to any one of claims 3 to 24, wherein the cancer expresses MAGE-A4 and / or is a solid tumor, and optionally, the solid tumor is urothelial cancer, head and neck cancer, non-small cell lung cancer (NSCLC), esophageal cancer, esophagogastric junction cancer, gastric cancer, ovarian cancer, melanoma, or endometrial cancer.

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