Methods for treating cancer or tumors

Abstract

The present invention relates to a method of treating, preventing, or delaying the progression of cancer and / or tumors in a subject, comprising administering to the subject a therapeutic regimen comprising an effective amount of modified immunoresponsive cells that express or display a heterologous T cell receptor (TCR) having the property of binding to MAGE A4.

Description

[Technical Field]

[0001] The present invention relates to a method for treating, inhibiting, or delaying the progression of cancer and / or tumors in a subject, comprising a therapeutic regimen comprising a step of applying to the treatment of gastroesophageal cancer, comprising an effective amount of modified immune-responsive cells expressing or presenting a heterologous T cell receptor (TCR) having the property of binding to MAGE A4 or its antigen peptide. [Background technology]

[0002] Gastric and esophageal cancer and / or tumors include cancer and / or tumors of the esophagus, gastroesophageal junction, or stomach, i.e., gastric cancer.

[0003] Esophageal cancer: Esophageal cancer is the sixth leading cause of cancer-related death worldwide. Generally, esophageal cancer arises from the epithelium of the esophagus and is classified into two classes: esophageal squamous cell carcinoma (ESCC), which is strongly associated with tobacco and alcohol consumption, and esophageal adenocarcinoma (EAC), which is commonly associated with GERD and Barrett's esophagus. Esophageal cancer includes esophagogastric junction cancer, or carcinoma or adenocarcinoma (EGJ), which is cancer of the lower part of the esophagus and is often associated with Barrett's esophagus. EGJ is a highly mutated and heterogeneous disease with an increased number of somatic mutations across several genes, such as CR2, HGF, FGFR4, ESRRB, TP53, SYNE1, and ARID1A. Treatment options for esophagogastric junction adenocarcinoma are limited, and the overall prognosis is extremely poor. ESCC (esophageal squamous cell carcinoma) accounts for 60–70% of all esophageal cancer cases worldwide, with another 20–30% being EAC (esophageal adenocarcinoma). Other less common forms of this cancer include melanoma, leiomyosarcoma, carcinoid, and lymphoma. Generally, the prognosis for esophageal cancer is quite poor, with an overall 5-year survival rate of approximately 15% in the United States, and most people die within the first year of diagnosis. Recent data from England and Wales show that only 10% of people survive esophageal cancer for at least 10 years. The poor prognosis explains the prevalence of the disease, and the low survival rate is also linked to a low rate of early detection, as most patients present with advanced disease by the time they experience the first symptoms, such as difficulty swallowing. In the United States, esophageal cancer is the seventh leading cause of cancer death in men. Curative treatment options for localized disease include a combination of surgery, radiation, and chemotherapy.

[0004] Metastatic or recurrent disease is generally managed temporarily with radiotherapy and chemotherapy, along with stent placement to alleviate symptoms and facilitate swallowing.

[0005] Gastric and stomach cancers are closely linked to tobacco, alcohol, and H. pylori, as well as salty or pickled foods. Treatment generally includes chemotherapy (with drugs such as 5-fluorouracil, cisplatin, epirubicin, etoposide, docetaxel, oxaliplatin, capecitabine, or irinotecan), radiotherapy, or surgery combined with targeted therapy, such as treatment with the human epidermal growth factor receptor 2 (HER2) inhibitor trastuzumab, which is overexpressed in 13-22% of patients with gastric cancer. While some types of gastric lymphoma can be cured by eliminating H. pylori, the prognosis is generally poor, mainly because most people with this condition present with progressive disease. Globally, stomach cancer is the third leading cause of cancer death, occurring twice as often in men as in women, accounting for 9% of cancer deaths in the United States, with a 5-year survival rate of 31.5%.

[0006] Therefore, it is desirable to provide a therapy for tumors and / or cancer treatment, such as the treatment of cancers and / or tumors of the esophagus, gastroesophageal junction, or gastroesophageal cancers and / or tumors, the therapy being cancer-specific and capable of treating intermediate or late-stage cancers or single or multiple solid tumors that have failed or recurred, particularly after first-line therapy or surgery, and preferably the therapy further minimizes or reduces the risk of toxicity or side effects, such as systemic toxicity of chemotherapeutic agents (e.g., nausea, vomiting, anemia, and thrombocytopenia) or tissue damage caused by radiotherapy.

[0007] The present invention relates to and illustrates a therapy for gastroesophageal cancer and / or tumors in a subject, comprising the step of administering a therapeutic regimen to the subject that comprises an effective amount of modified T cells expressing or presenting a heterologous T cell receptor (TCR) having the property of binding to MAGE A4, in particular GVYDGREHTV, SEQ ID NO: 2; in particular, GVYDGREHTV, SEQ ID NO: 2, an HLA-A2 restricted MAGE A4 peptide that provides a suitable target for a novel immunotherapeutic intervention; this peptide is naturally processed and isolated from gastroesophageal cancer cell lines. [Overview of the Initiative]

[0008] According to a first aspect of the present invention, a method is provided for treating, inhibiting, or delaying the progression of gastroesophageal cancer and / or tumor in a subject, comprising the step of administering a therapeutic regimen to the subject comprising an effective amount of modified immune-responsive cells expressing or presenting a heterologous T cell receptor (TCR) or chimeric antigen receptor (CAR) bound to MAGE A4 or its MAGE A4 antigen peptide.

[0009] According to the present invention, TCR or CAR can bind to MAGE A4 or its antigenic peptide, for example, human MAGE A4 or MAGE A4 of Sequence ID No. 1 or its antigenic peptide. TCR or CAR can bind to antigenic peptides including Sequence ID No. 2, GVYDGREHTV.

[0010] In one embodiment, the present invention provides modified immune-responsive cells expressing or presenting a MAGE A4 peptide antigen or a heterologous T cell receptor (TCR) or CAR that binds to the antigen peptide, for use in treating, inhibiting, or delaying the progression of cancer and / or tumors in a subject, wherein the cancer and / or tumor is gastroesophageal cancer and / or tumor.

[0011] In embodiments, the TCR or CAR may bind to human MAGE A4 or MAGE A4 of SEQ ID NO: 1 or its antigenic peptide. The TCR or CAR may bind to antigenic peptides including SEQ ID NO: 2, GVYDGREHTV.

[0012] In this embodiment, the cells are administered to a subject in a therapeutic regimen containing an effective amount of the cells. [Brief explanation of the drawing]

[0013] [Figure 1] These CT scan (computed tomography) data demonstrate a >42% reduction in target lesions in esophagogastric junction (EGJ) cancer over a 12-week period following MAGE-A4 CD8 T cell infusion in a cohort of subjects who received approximately 10 billion cells. [Figure 2] This table shows the tumor response for esophagogastric junction (EGJ) cancer after MAGE-A4 CD8 T cell infusion, representing the percentage change in the sum of diameters (sum of diameters = sum of the long axis for non-narrow lesions and the short axis for nodular lesions) from baseline lesion SLC, i.e., the percentage change in the sum of diameters in the target lesion measurement. The response was evaluated according to RECIST v1.1. [Modes for carrying out the invention]

[0014] Immune-responsive cells According to the present invention, modified immune-responsive cells may be lymphoid cells, including B, T, or natural killer (NK) cells. Modified immune-responsive cells may be lymphoid cells, including T cells, natural killer T (NKT) cells, and their precursors, including embryonic stem cells and pluripotent stem cells (e.g., from which lymphoid cells can differentiate). T cells may be lymphocytes that mature in the thymus and are primarily involved in cell-mediated immunity and also in the adaptive immune system. According to the present invention, T cells may include, but are not limited to, helper T cells, cytotoxic T cells, memory T cells (including central memory T cells, stem cell-like memory T cells (or stem-like memory T cells), and two types of effector memory T cells: e.g., TEM cells and TEMRA cells), regulatory T cells (also known as suppressor T cells), natural killer T cells, mucosa-associated invariant T cells, and gamma-delta T cells. Cytotoxic T cells (CTLs or killer T cells) are a subset of T lymphocytes capable of inducing the death of infected somatic or tumor cells. The subject's own T cells can be genetically modified to target specific antigens by introducing xenolytic TCRs or CARs. Preferably, the modified immune-responsive cells are T cells, optionally CD4 + T cells or CD8 + These are T cells. Therefore, modified immune-responsive cells may be T cells, optionally CD4+ T cells or CD8+ T cells, or modified immune-responsive cells may be a population of modified T cells, optionally CD4+ T cells; or CD8+ T cells, or a mixed population of CD4+ T cells and CD8+ T cells.

[0015] Heterogeneous TCR / CAR According to the present invention, modified immunoreactive cells can express a heterologous T cell receptor (TCR) or a heterologous chimeric antigen receptor (CAR) (e.g., the cells are engineered to be transduced with or to contain a nucleic acid sequence encoding a heterologous TCR or CAR, such as by gene knock-in). When bound to an antigen, the modified immunoreactive cells can exhibit T cell effector functions and / or a cytolytic effect against cells bearing the antigen and / or can undergo proliferation and / or cell division. In certain embodiments, modified immunoreactive cells comprising a heterologous TCR exhibit comparable or superior therapeutic efficacy compared to cells comprising a chimeric antigen receptor (CAR) that targets the same cancer and / or tumor antigen and / or peptide (antigenic peptide). Activated modified immunoreactive cells comprising a heterologous TCR or CAR can secrete anti-tumor cytokines, including but not limited to TNF alpha, IFNγ, and IL2.

[0016] According to the present invention, modified immunoreactive cells can contain a nucleic acid, construct, or vector encoding a heterologous T cell receptor (TCR) or a heterologous chimeric antigen receptor (CAR), or a heterologous nucleic acid, construct, or vector. Optionally, the TCR can be an affinity-enhanced TCR, such as a specific peptide-enhanced affinity receptor (SPEAR) TCR.

[0017] The terms "heterologous" or "exogenous" refer to a polypeptide or nucleic acid that is foreign to a particular biological system, such as a cell or host cell, does not naturally occur in that system, and can be introduced into the system by artificial or recombinant means. Thus, the expression of a heterologous TCR or CAR can thereby alter the immunogenic specificity of immunogenic cells, such as T cells, such that they recognize or exhibit enhanced recognition of one or more tumor or cancer antigens and / or peptides present on the surface of cancer cells of an individual having cancer. Modification of immunogenic cells or T cells and their subsequent expansion can be performed in vitro and / or ex vivo.

[0018] Cancer / tumor antigen or peptide antigen According to the present invention, the cancer and / or tumor antigen or its peptide antigen can be a cancer-testis antigen such as MAGE, melanoma-associated antigen, or a member of the MAGEA gene family, for example, MAGE A1, A2, A3, A4, A5, A6, A7, A8, A9, A10, A11, or A12, or any one of their peptide antigens. Preferably, the tumor antigen is MAGE-A4, SEQ ID NO: 1, or its peptide antigen. Preferably, the cancer and / or tumor antigen peptide contains or has the amino acid sequence GVYDGREHTV, SEQ ID NO: 2.

[0019] Costimulatory ligand According to the present invention, modified immune-responsive cells may further comprise at least one, optionally one, two, three, or four, co-stimulatory ligands, which are exogenous or recombinant (for example, cells may be transduced or manipulated to contain a nucleic acid sequence encoding a co-stimulatory ligand, e.g., by gene knock-in). Modified immune-responsive cells may co-express a heterologous TCR or CAR and at least one exogenous or heterologous co-stimulatory ligand. The interaction between the heterologous TCR or CAR and at least one exogenous co-stimulatory ligand may provide non-antigen-specific signaling and / or cell activation. Co-stimulatory ligands include, but are not limited to, members of the tumor necrosis factor (TNF) superfamily and immunoglobulin (Ig) superfamily ligands. TNF is a cytokine involved in systemic inflammation and stimulates the acute phase response. The TNF superfamily members include, but are not limited to, nerve growth factor (NGF), CD40L (CD40L) / CD154, CD137L / 4-1BBL, TNF-alpha, CD134L / OX40L / CD252, CD27L / CD70, Fas ligand (FasL), CD30L / CD153, tumor necrosis factor beta (TNFP) / lymphotoxin-alpha (LTa), lymphotoxin-beta (TTb), CD257 / B cell activator (BAFF) / Blys / THANK / Tall-I, glucocorticoid-inducible TNF receptor ligand (GITRL), and TNF-related apoptosis-inducing ligand (TRAIL), LIGHT (TNFSF14). The immunoglobulin (Ig) superfamily is a large group of soluble proteins on the cell surface involved in cell recognition, binding, or adhesion processes. These proteins share structural characteristics with immunoglobulins, and they possess an immunoglobulin domain (fold). Immunoglobulin superfamily ligands include, but are not limited to, CD80 and CD86, both of which are ligands for CD28. In certain embodiments, at least one co-stimulatory ligand is selected from the group consisting of 4-1BBL, CD275, CD80, CD86, CD70, OX40L, CD48, TNFRSF14, and combinations thereof.At least one exogenous or recombinant costimulatory ligand may be 4-1BBL or CD80, preferably at least one exogenous or recombinant costimulatory ligand is 4-1BBL. Modified immune-responsive cells may contain two exogenous or recombinant costimulatory ligands, preferably the two exogenous or recombinant costimulatory ligands are 4-1BBL and CD80.

[0020] Modified immune-responsive cells may comprise at least one exogenous or recombinant construct (e.g., cells are transduced or engineered to express, e.g., by gene knock-in) that overcomes the immunosuppressive tumor microenvironment. Such constructs may include, but are not limited to, cyclic AMP phosphodiesterase and dominant-negative transforming growth factor beta (TGF beta) receptor II. Modified immune-responsive cells, modified T cells, or populations of modified T cells may be engineered to release cytokines that have a positive effect on the cytolytic activity of the cells. Such cytokines may include, but are not limited to, interleukin-7, interleukin-15, and interleukin-21.

[0021] Specific binding TCR / CAR According to the present invention, modified immune-responsive cells, such as modified T cells, may optionally be modified to express tumor cells and / or tissues of a subject, patient, or cancer patient suffering from a diseased or cancerous condition, as well as heterologous TCRs or CARs that bind to or specifically bind to cancer cells and / or tissues, expressing or presenting cancer and / or tumor antigens or their peptide antigens as described herein. The subject, patient, or cancer patient may then be treated with modified immune-responsive cells or modified T cells or populations thereof in accordance with the present invention. Cancer patients suitable for treatment according to the present invention using modified immune-responsive cells or modified T cells may be identified by a method comprising the steps of: obtaining a sample of tumor and / or cancer cells from an individual or subject having tumor and / or cancer; and identifying cancer cells as binding to TCRs or CARs expressed by modified immune-responsive cells.

[0022] According to the present invention, heterologous TCRs or CARs bind to or specifically bind to cancer and / or tumor antigens or their peptide antigens, which are associated with a cancerous condition and / or presented by a tumor or cancer cells or tissues.

[0023] According to the present invention, the cancerous state may be gastroesophageal cancer and / or tumor. Specificity describes the strength of binding between a heterologous TCR or CAR and a specific target cancer and / or tumor antigen or its peptide antigen, and can be described by a dissociation constant Kd, which is the ratio between bound and unbound states in the receptor-ligand system. Additionally, the fewer different cancer and / or tumor antigens or their peptide antigens to which a heterologous TCR or CAR can bind, the greater its binding specificity.

[0024] According to the present invention, heterologous TCRs or CARs can bind to 10, 9, 8, 7, 6, 5, 4, 3, or fewer than 2 different cancer and / or tumor antigens or their peptide antigens.

[0025] According to the present invention, heterogeneous TCR or CAR is measured at an optional pH of 6.5-6.9 or 7.0-7.5, optionally at 25°C, and optionally using surface plasmon resonance, in the following ranges: 0.01 μM-100 μM, 0.01 μM-50 μM, 0.01 μM-20 μM, 0.05 μM-20 μM, or 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1 μM, 0.15 μM, 0.2 μM, 0.25 μM, 0.3 μM, 0.35 μM, 0.4 μM, 0.45 μM, 0.5 μM, 0.55 μM, 0.6 μM, 0.65 μM, 0.7 μM, 0.75 μM, 0. 8μM, 0.85μM, 0.9μM, 0.95μM, 1.0μM, 1.5μM, 2.0μM, 2.5μM, 3.0μM, 3.5μM, 4.0μM, 4. 5μM, 5.0μM, 5.5μM, 6.0μM, 6.5μM, 7.0μM, 7.5μM, 8.0μM, 8.5μM, 9.0μM, 9.5μM, 10.0μM M; or dissociation constants of 10 μM to 1000 μM, 10 μM to 500 μM, 50 μM to 500 μM, or 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 μM, 150 μM, 200 μM, 250 μM, 300 μM, 350 μM, 400 μM, 450 μM, 500 μM, which can bind to, for example, MAGE A4 or its antigen peptide, for example, human MAGE A4 or MAGE A4 of SEQ ID NO: 1 or its antigen peptide, or antigen peptides containing or consisting of SEQ ID NO: 2, GVYDGREHTV. Dissociation constant K D or k off / k on The dissociation rate constant k off and the association rate constant k onThis can be determined by experimental measurement. The TCR dissociation constant can be measured using the soluble form of the TCR, which includes the TCR alpha-chain variable domain and the TCR beta-chain variable domain. Thus, heterologous TCRs or CARs for use according to the present invention can efficiently and / or bind with high affinity to HLA presenting GVYDGREHTV, SEQ ID NO: 2, by optionally forming complexes with peptide-presenting molecules, such as HLA, for example, HLA-A*02 or HLA-A*0201, or alternatively without complexing with peptide-presenting molecules.

[0026] According to the present invention, modified immune-responsive cells, such as modified T cells, may optionally include heterologous TCRs or CARs that are associated with a cancerous state and / or presented by a tumor or cancer cells or tissue, and that can specifically bind to and / or bind with high affinity to cancer and / or tumor antigens or their peptide antigens; optionally, cancer and / or tumor antigens or their peptide antigens may be recognized by heterologous TCRs or CARs by optionally forming a complex with a peptide-presenting molecule, such as HLA, such as HLA-A*02 or HLA-A*0201, or alternatively, without presentation that forms a complex with a peptide-presenting molecule, such as HLA (i.e., MAGE-A4 or its peptide antigen, or the peptide antigen of MAGE A4 including GVYDGREHTV, SEQ ID NO: 2, may be presented independently of a peptide-presenting molecule). For example, cancer and / or tumor antigens or their peptide antigens are antigen peptides containing or consisting of MAGE A4 or its antigenic peptide, such as human MAGE A4 or MAGE A4 of Sequence ID No. 1 or its antigenic peptide, or Sequence ID No. 2, GVYDGREHTV.

[0027] According to the present invention, heterologous T cell receptors (TCRs) or CARs, and modified immune-responsive cells containing heterologous T cell receptors (TCRs) or CARs, may have the property of binding to endogenously expressed tumor cell surfaces, cancer and / or tumor antigens or their peptide antigens, and optionally, the binding is independent of the presentation of cell surface antigens as a complex with peptide-presenting or antigen-presenting molecules, such as major histocompatibility complex (MHC) or human leukocyte antigen (HLA) or major histocompatibility complex class-associated protein (MR) 1. For example, the cancer and / or tumor antigen or its peptide antigen is an antigen peptide containing or consisting of MAGE A4 or its antigen peptide, such as human MAGE A4 or MAGE A4 of SEQ ID NO: 1, or SEQ ID NO: 2, GVYDGREHTV.

[0028] According to the present invention, the TCR or CAR binding may optionally be specific to one cancer and / or tumor antigen, such as human MAGE A4 or MAGE A4 of SEQ ID NO: 1, or its antigenic peptide, or to an antigenic peptide containing or consisting of SEQ ID NO: 2, GVYDGREHTV, compared to a closely related cancer and / or tumor antigen or peptide antigen sequence. The closely related cancer and / or tumor antigen or peptide antigen sequences may be of similar or identical length and / or have a similar or identical number of amino acid residues. The closely related peptide antigen sequences may share 50, 60, 70, or 80-90% identity, preferably 80-90% identity, and / or differ by only 1, 2, 3, or 4 amino acid residues. The closely related peptide sequence may contain the sequence or be derived from a polypeptide sequence having the sequence GVYDGREHTV, SEQ ID NO: 2.

[0029] Binding affinity can be determined by equilibrium methods (e.g., enzyme-linked immunosorbent assay (ELISA) or radioimmunoassay (RIA)) or kinetics (e.g., BIACORE® analysis). Binding strength is the sum of the strengths of binding between two molecules at multiple sites, taking into account, for example, the valence of the interaction. According to the present invention, immune-responsive cells may exhibit improved affinity and / or binding strength to cancer and / or tumor antigens or their peptide antigens, or cancer and / or tumor antigens or their peptide antigens presented by tumors or cancer cells or tissues and recognized by heterologous TCRs or CARs, compared to immune-responsive cells lacking or having alternative heterologous TCRs or CARs.

[0030] Selective binding TCR / CAR According to the present invention, heterologous TCRs or CARs may selectively bind to cancer and / or tumor antigens or their peptide antigens that are optionally associated with a cancerous condition and / or presented by a tumor or cancer cells or tissue; optionally, the cancer and / or tumor antigens or their peptide antigens may optionally bind to peptide-presenting molecules, such as major histocompatibility complexes (MHC) or HLA, optionally class I or II, such as HLA-A2 or HLA-A*O2, HLA-A*O2:0 1. Forms a complex with HLA-A*02:02, HLA-A*02:03, HLA-A*02:04, HLA-A*02:05, HLA-A*02:06, HLA-A*02:642, or HLA-A*02:07, preferably selected from HLA-A*02:01 or HLA-A*0; alternatively, is recognized by heterologous TCRs or CARs without presentation, preferably without forming a complex with peptide-presenting molecules or HLA expressed by tumor cells or cancer cells or tissues. Preferably, the malignant condition is gastroesophageal cancer and / or tumor.

[0031] Selective binding refers to the binding of a heterologous TCR or CAR to a particular cancer and / or tumor antigen or its peptide antigen with higher affinity than to another. Selective binding is represented by the equilibrium constant for the substitution of one ligand antigen by the other ligand antigen in the complex with the heterologous TCR or CAR.

[0032] According to the present invention, the cancerous condition may be gastroesophageal cancer and / or tumor.

[0033] Specific / selective binding of TCRs / CARs According to the present invention, heterologous TCR or CAR binding is selective and / or specific to cancer and / or tumor antigens or their peptide antigens, which may be MAGE-A4 or its peptide antigen. Preferably, the tumor antigen is MAGE-A4 or its peptide antigen. Preferably, the cancer and / or tumor antigen peptide contains or has the amino acid sequence GVYDGREHTV, SEQ ID NO: 2. According to the present invention, heterologous TCRs or CARs can bind to peptide-presenting molecules, such as cancer and / or tumor antigens or HLAs that present or display such peptide antigens, i.e., peptide fragments (pHLAs) of cancer and / or tumor antigens, and can bind specifically and / or selectively, wherein the HLAs correspond to MHC class I (A, B, and C), all of which are HLA class I or their specific alleles, or the HLAs correspond to MHC class II (DP, DM, DO, DQ, and DR) or their specific alleles, preferably the HLAs are class I, and preferably the alleles are HLA-A2 or HLA-A*02 or HLA-A2+ or HLA-A*02 positive HLA, preferably HLA-*0201. Preferably the HLAs are neither HLA-A*02:07P nor HLA-A*02 null. Alternatively, heterologous TCRs or CARs may bind to cancer and / or tumor antigens or their peptide antigens that are not presented or expressed by HLA, and / or may bind specifically and / or may bind selectively.

[0034] Preferably, the heterologous TCR or CAR is not spontaneously expressed by immune-responsive cells (i.e., the TCR or CAR is exogenous or heterologous). The heterologous TCR may include an αβ TCR heterodimer. The heterologous TCR or CAR may be a recombinant, synthetic, or artificial TCR or CAR, i.e., a CAR or TCR that does not exist in nature. For example, the heterologous TCR may be engineered to increase its affinity or binding affinity to specific cancer and / or tumor antigens or their peptide antigens (i.e., affinity-enhancing TCR or specific peptide-enhancing affinity receptor (SPEAR) TCR). The affinity-enhancing TCR or (SPEAR) TCR may contain one or more mutations compared to the naturally occurring TCR, for example, one or more mutations in the hypervariable complementarity-determining regions (CDRs) of the variable regions of the TCRα and β chains. These mutations, when expressed selectively by tumors and / or cancer cells and / or tissues, may increase the affinity of TCRs to peptide fragments of cancer and / or tumor antigens or their peptide antigens, or to MHCs presenting peptide fragments of cancer and / or tumor antigens. Appropriate methods for affinity enhancement or the creation of mature TCRs are well known in the art and involve a step of screening a library of TCR variants using phage or yeast display (see, for example, Robbins et al J Immunol (2008) 180(9):6116; San Miguel et al (2015) Cancer Cell 28 (3) 281-283; Schmitt et al (2013) Blood 122 348-256; Jiang et al (2015) Cancer Discovery 5 901). Preferred affinity-enhancing TCRs can bind to tumor or cancer cells expressing cancer and / or tumor antigens of the MAGE family, such as MAGE A4 or its peptide antigen, such as sequence GVYDGREHTV, SEQ ID NO: 2, or a peptide comprising the same.

[0035] According to the present invention, a heterologous TCR may be a MAGE A4 TCR that may include the α-chain reference amino acid sequence of SEQ ID NO: 5 or a variant thereof and the β-chain reference amino acid sequence of SEQ ID NO: 7 or a variant thereof. The variant may have an amino acid sequence that has sequence identity of at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% with respect to the reference amino acid sequence (for example, with respect to either the α-chain reference sequence and / or the β-chain reference sequence). The TCR may be encoded by the α-chain reference nucleotide sequence of SEQ ID NO: 6 or a variant thereof and the β-chain reference nucleotide sequence of SEQ ID NO: 8 or a variant thereof. The variant may have a nucleotide sequence that has at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity with respect to the reference nucleotide sequence (for example, with respect to either the α-chain reference sequence and / or the β-chain reference sequence).

[0036] According to the present invention, the TCR may include a TCR alpha chain variable domain and a TCR beta chain variable domain. (i) The alpha chain variable domain is sequence VSPFSN(αCDR1), SEQ ID NO: 11, or amino acid residues 48-53 of SEQ ID NO: 5 LTFSEN(αCDR2), amino acid residues 71-76 of SEQ ID NO: 12, or SEQ ID NO: 5, and CVVSGGTDSWGKLQF(αCDR3), SEQ ID NO: 13, or amino acid residues 111-125 of SEQ ID NO: 5 Includes a CDR having and / or (ii) The beta chain variable domain is sequence KGHDR(βCDR1), amino acid residues 46-50 of SEQ ID NO: 14, or SEQ ID NO: 7, SFDVKD(βCDR2), amino acid residues 68-73 of SEQ ID NO: 15, or SEQ ID NO: 7, and CATSGQGAYEEQFF(βCDR3), amino acid residues 110-123 of SEQ ID NO: 16, or SEQ ID NO: 7, or sequences with at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity, or optionally sequences with 100% sequence identity. Includes a CD-R containing [specific feature].

[0037] Therefore, a TCR may include one in which the alpha-chain variable domain contains an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 9, or the sequence of amino acid residues 1-136 of SEQ ID NO: 6, and / or the beta-chain variable domain contains an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 10, or the sequence of amino acid residues 1-133 of SEQ ID NO: 7.

[0038] The terms “parent TCR” or “precursor TCR” are used herein to refer to TCRs comprising the MAGE A4 TCR α chain and MAGE A4 TCR β chain of Sequence ID No. 5 and 7, respectively. It is desirable to provide a mutated or modified TCR compared to the precursor TCR that has equal, equivalent, or higher affinity and / or equal, equivalent, or slower off-rate to the peptide-HLA complex than the precursor TCR. According to the present invention, heterologous TCRs may have one or more mutations present in the alpha chain variable domain and / or beta chain variable domain compared to the precursor TCR, and may be referred to as “engineered TCRs” or “mutant TCRs.” These mutations may improve binding affinity and / or specificity and / or selectivity and / or binding strength to MAGE A4 or its peptide antigen. In certain embodiments, there may be 1, 2, 3, 4, 5, 6, 7, or 8 mutations in the alpha chain variable domain, e.g., 4 or 8 mutations, and / or 1, 2, 3, 4, or 5 mutations in the beta chain variable domain, e.g., 5 mutations. In some embodiments, the α-chain variable domain of the TCR of the present invention may include an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the amino acid residue sequence of SEQ ID NO: 9. In some embodiments, the β-chain variable domain of the TCR of the present invention may include an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the amino acid residue sequence of SEQ ID NO: 10.

[0039] According to the present invention, the heterologous TCR has an alpha chain variable domain in which the amino acid sequence of amino acid residues 1-136 of SEQ ID NO: 9 or SEQ ID NO: 5, or the amino acid residues 1-47, 54-70, 77-110, and 126-136 of SEQ ID NO: 9 are at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 9% of the amino acid sequence of amino acid residues 1-47, 54-70, 77-110, and 126-136 of SEQ ID NO: 9. The TCR may include an amino acid sequence having 9% identity and / or where amino acid residues 48-53, 71-76, and 111-125, CDR1, CDR2, and CDR3 respectively have at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with the sequences of amino acid residues 48-53, 71-76, and 111-125, CDR1, CDR2, and CDR3 of SEQ ID NO: 9.

[0040] According to the present invention, the TCR, in the alpha chain variable domain, has the following sequence: (i) The amino acid residues 1-47 may (a) have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 1-47 of SEQ ID NO: 9, or (b) may have 1, 2, or 3 amino acid residues inserted or deleted compared to residues 1-47 of SEQ ID NO: 9. (ii) Amino acid residues 48-53 are amino acid residues 48-53 of VSPFSN, CDR1, SEQ ID NO: 11, or SEQ ID NO: 9, (iii) The amino acid residues 54-70 may (a) have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 54-70 of SEQ ID NO: 9, or (b) may have one, two, or three amino acid residues inserted or deleted compared to the sequence of amino acid residues 54-70 of SEQ ID NO: 9. (iv) Amino acid residues 71-76 may be amino acid residues 71-76 of LTFSEN, CDR2, SEQ ID NO: 12, or SEQ ID NO: 9, (v) The amino acid residues 77-110 may have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 77-110 of SEQ ID NO: 9, or may have 1, 2, or 3 insertions, deletions, or substitutions compared to the sequence of amino acid residues 77-110 of SEQ ID NO: 9 (vi) Amino acid residues 111-125 may be CVVSGGTDSWGKLQF, CDR3, SEQ ID NO: 13, or SEQ ID NO: 9, (vii) The amino acid residues 126-136 may have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 126-136 of SEQ ID NO: 9, or may have 1, 2, or 3 insertions, deletions, or substitutions compared to the sequence of amino acid residues 126-136 of SEQ ID NO: 9 May contain TCRs.

[0041] According to the present invention, a TCR may include an amino acid sequence in which the beta-chain variable domain has at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the amino acid sequence of SEQ ID NO: 10, or its amino acid residues 1-45, 51-67, 74-109, and 124-133, respectively, of the sequences of amino acid residues 1-45, 51-67, 74-109, and 124-133, respectively, and amino acid residues 46-50, 68-73, and 110-123, respectively, of the sequences of amino acid residues 46-50, 68-73, and 110-123, respectively, of CDR1, CDR2, and CDR3.

[0042] According to the present invention, the TCR has a sequence in the beta chain variable domain: (i) The amino acid residues 1-45 may (a) have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 1-45 of SEQ ID NO: 10, or (b) may have 1, 2, or 3 amino acid residues inserted or deleted compared to residues 1-45 of SEQ ID NO: 10. (ii) Amino acid residues 46-50 are amino acid residues 46-50 of KGHDR, CDR1, SEQ ID NO: 14, or SEQ ID NO: 10, (iii) The amino acid residues 51-67 may (a) have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 51-67 of SEQ ID NO: 10, or (b) have one, two, or three amino acid residues inserted or deleted compared to the sequence of amino acid residues 51-67 of SEQ ID NO: 10. (iv) Amino acid residues 68-73 may be amino acid residues 68-73 of SFDVKD, CDR2, SEQ ID NO: 15, or SEQ ID NO: 10, (v) The amino acid residues 74-109 may have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 74-109 of SEQ ID NO: 10, or may have 1, 2, or 3 insertions, deletions, or substitutions compared to the sequence of amino acid residues 74-109 of SEQ ID NO: 10. (vi) Amino acid residues 110-123 may be amino acid residues 110-123 of CATSGQGAYEEQFF, CDR3, SEQ ID NO: 16, or SEQ ID NO: 10, (vii) The amino acid residues 124-133 may have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 124-133 of SEQ ID NO: 10, or may have 1, 2, or 3 insertions, deletions, or substitutions compared to the sequence of amino acid residues 124-133 of SEQ ID NO: 10. May contain TCRs.

[0043] According to the present invention, a TCR may include a TCR comprising the alpha chain variable domain of SEQ ID NO: 9 and / or the beta chain variable domain of SEQ ID NO: 10. According to the present invention, a TCR may include a TCR comprising the alpha chain of SEQ ID NO: 5 and / or the beta chain of SEQ ID NO: 7.

[0044] Amino acid and nucleotide sequence identity is generally defined by the GAP algorithm (GCG Wisconsin Package®, Accelrys, San Diego, CA). GAP aligns two complete sequences using the Needleman & Wunsch algorithm (J. Mol. Biol. (48): 444-453 (1970)), which maximizes the number of matches and minimizes the number of gaps. Typically, initial parameters with a gap creation penalty of 12 and a gap extension penalty of 4 are used. While the use of GAP may be preferred, other algorithms may be used, such as BLAST, psiBLAST, or TBLASTN (using the method described in Altschul et al. (1990) J. Mol. Biol. 215: 405-410), FASTA (using the method described in Pearson and Lipman (1988) PNAS USA 85: 2444-2448), or the Smith-Waterman algorithm (Smith and Waterman (1981) J. Mol Biol. 147: 195-197), which generally employ default parameters.

[0045] A particular amino acid variant may differ from the reference sequence by one, two, three, four, five to ten, ten to twenty, or twenty to thirty amino acid insertions, additions, substitutions, or deletions. In some embodiments, the variant sequence may include a reference sequence having one, two, three, four, five, six, seven, eight, nine, ten, or more inserted, deleted, or substituted residues. For example, up to 15, up to 20, up to 30, or up to 40 residues may be inserted, deleted, or substituted.

[0046] In some preferred embodiments, the variant may differ from the reference sequence by one, two, three, four, five, six, seven, eight, nine, ten, or more conserved substitutions. Conservative substitutions involve the replacement of an amino acid with a different amino acid having similar properties. For example, an aliphatic residue may be replaced by another aliphatic residue, a nonpolar residue by another nonpolar residue, an acidic residue by another acidic residue, a basic residue by another basic residue, a polar residue by another polar residue, or an aromatic residue by another aromatic residue. Conservative substitutions may, for example, be between amino acids within the following group: Alanine and glycine; Glutamic acid, aspartic acid, glutamine, and asparagine Arginine and lysine; Asparagine, glutamine, glutamic acid, and aspartic acid Isoleucine, leucine, and valine; Phenylalanine, tyrosine, and tryptophan Serine, threonine, and cysteine.

[0047] CD8α coreceptor According to the present invention, modified immune-responsive cells expressing or presenting a heterologous TCR or CAR may further express or present a heterologous coreceptor (for example, cells may be transduced or manipulated to include a nucleic acid sequence encoding the coreceptor, for example, by gene knock-in). The heterologous coreceptor may be a CD8 coreceptor. The CD8 coreceptor may include a dimer or pair of CD8 chains, comprising CD8-α and CD8-β chains or CD8-α and CD8-α chains. Preferably, the CD8 coreceptor is a CD8αα coreceptor comprising CD8-α and CD8-α chains. The CD8α coreceptor may include an amino acid sequence, SEQ ID NO: 3, or a variant thereof, that is at least 80% identical to SEQ ID NO: 3. The CD8α coreceptor may be a homodimer. Preferably, modified immune-responsive cells expressing or presenting a heterologous TCR or CAR further express or present a heterologous coreceptor, which is a CD8 coreceptor as defined herein (for example, cells are transduced or manipulated to include a nucleic acid sequence encoding the coreceptor, for example, by gene knock-in).

[0048] The CD8 coreceptor binds to class 1 MHC and enhances TCR signaling. According to the present invention, the CD8 coreceptor may include the reference amino acid sequence of SEQ ID NO: 3, or a variant thereof. The variant may have an amino acid sequence having at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity with the reference amino acid sequence of SEQ ID NO: 3. The CD8 coreceptor may be encoded by the reference nucleotide sequence of SEQ ID NO: 4, or a variant thereof. The variant may have a nucleotide sequence that has at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity with reference nucleotide sequence number 4.

[0049] According to the present invention, heterologous CD8 coreceptors have an Ig-like V-type domain with a sequence; (i) Amino acid residues 45-53 of VLLSNPTSG, CDR1, SEQ ID NO: 17, or SEQ ID NO: 3, (ii) amino acid residues 72-79 of YLSQNKPK, CDR2, SEQ ID NO: 18, or SEQ ID NO: 3, (iii) amino acid residues 80-117 of LSNSIM, CDR3, SEQ ID NO: 19, or SEQ ID NO: 3, Alternatively, a sequence having at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or 99% sequence identity with it. It may include CD8 coreceptors, including CDRs that have the following properties.

[0050] According to the present invention, a heterologous CD8 coreceptor may include a CD8 coreceptor in which amino acid residues 22-135 of the amino acid sequence of SEQ ID NO: 3, or amino acid residues 22-44, 54-71, 80-117, and 124-135, have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequences of amino acid residues 22-44, 54-71, 80-117, 124-135, CDR1, CDR2, and CDR3 of SEQ ID NO: 3, and amino acid residues 45-53, 72-79, and 118-123 have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequences of amino acid residues 45-53, 72-79, and 118-123 of SEQ ID NO: 3, or are included in an Ig-like V-type domain.

[0051] According to the present invention, the CD8 coreceptor has a sequence, or a sequence in the Ig-like V domain: (i) The amino acid residues 22-44 may (a) have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 22-44 of SEQ ID NO: 3, or (b) may have one, two, or three amino acid residues inserted or deleted compared to residues 22-44 of SEQ ID NO: 3. (ii) Amino acid residues 45-53 are amino acid residues 45-53 of VLLSNPTSG, SEQ ID NO: 17, CDR1, or SEQ ID NO: 3, (iii) The amino acid residues 54-71 may (a) have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 54-71 of SEQ ID NO: 3, or (b) may have one, two, or three amino acid residues inserted or deleted compared to the sequence of amino acid residues 54-71 of SEQ ID NO: 3. (iv) Amino acid residues 72-79 may be YLSQNKPK, CDR2, SEQ ID NO: 18, or SEQ ID NO: 3, (v) The amino acid residues 80-117 may have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 80-117 of SEQ ID NO: 3, or may have one, two, or three insertions, deletions, or substitutions compared to the sequence of amino acid residues 80-117 of SEQ ID NO: 3; (vi) Amino acid residues 118-123 may be amino acid residues 80-117 of LSNSIM, CDR3, SEQ ID NO: 19, or SEQ ID NO: 3, (vii) The amino acid residues 124-135 may have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 124-135 of SEQ ID NO: 3, or may have 1, 2, or 3 insertions, deletions, or substitutions compared to the sequence of amino acid residues 124-135 of SEQ ID NO: 3. May include CD8 coreceptors.

[0052] Modified immune-responsive cells expressing heterologous CD8 coreceptors may exhibit improved affinity and / or binding affinity, as determined by assays disclosed herein, and / or improved T cell activation to or in response to antigen peptides, tumor or cancer antigens, when selectively presented on HLA, as determined by assays disclosed herein, compared with modified immune-responsive cells that do not express heterologous CD8 coreceptors. Heterologous CD8 in modified immune-responsive cells may interact with or specifically bind to MHC, which may be class I or class II, preferably class I major histocompatibility complex (MHC), HLA-I molecules, or MHC class I HLA-A / B2M dimers, and preferably CD8-α interacts with the α3 portion (residues 223-229) of class I MHC, preferably via the IgV-like domain of CD8. Therefore, heterologous CD8 enhances TCR binding of immune-responsive cells to HLA and / or antigen peptides bound or presented by HLA pMHCI or pHLA on the surface of antigen-presenting cells, dendritic cells, and / or tumor or cancer cells, or tumor or cancer tissue, compared to immune-responsive cells lacking heterologous CD8. Therefore, heterologous CD8 enhances the off-rate (k) of cell (TCR) / peptide-major histocompatibility complex class I (pMHCI) interactions of immune-responsive cells on the surface of antigen-presenting cells, dendritic cells, and / or tumor or cancer cells, or tumor or cancer tissue, compared to cells lacking heterologous CD8. off Therefore, its half-life may be improved or increased, thereby also providing improved ligation affinity and / or binding affinity. Heterogeneous CD8 may improve the organization of TCRs on the surface of immune-responsive cells, enabling cooperability in pHLA binding and potentially providing improved therapeutic binding affinity. Thus, heterogeneous CD8 coreceptor-modified immune-responsive cells may bind to or interact with LCK (lymphocyte-specific protein tyrosine kinase) in a zinc-dependent manner, leading to the activation of transcription factors such as NFAT, NF-κB, and AP-1.

[0053] According to the present invention, modified immune-responsive cells may, compared to immune-responsive cells lacking heterologous CD8 coreceptors, have improved or increased CD40L expression, cytokine production, cytotoxic activity, induction of dendritic cell maturation, or induction of dendritic cell cytokine production in an optional response to cancer and / or tumor antigens or their peptide antigens presented by cancer cells or tumor tissues.

[0054] therapy Gastric and esophageal cancer and / or tumors include cancer and / or tumors of the esophagus, gastroesophageal junction, or stomach, i.e., gastric cancer.

[0055] According to the present invention, cancer and / or tumor may be gastroesophageal cancer and / or tumor, for example, esophageal cancer, carcinoma, or tumor, which may be primary, secondary, recurrent, metastatic, or progressive. Preferably, esophageal cancer, carcinoma, or tumor may be optionally selected from any one of esophageal squamous cell carcinoma (ESCC), esophageal adenocarcinoma (EAC), or esophagogastric junction cancer, carcinoma, adenocarcinoma, or tumor (EGJ) associated with gastroesophageal reflux disease (GERD) or Barrett's esophagus.

[0056] Therefore, cancer and / or tumors may be gastroesophageal cancer and / or tumors, such as esophageal squamous cell carcinoma, or non-epithelial tumors such as leiomyosarcoma, malignant melanoma, rhabdomyosarcoma, or lymphoma.

[0057] Therefore, cancer and / or tumors may be gastroesophageal cancer and / or tumors, such as upper, middle, or lower junctional esophageal cancer, carcinoma, or tumor.

[0058] Therefore, cancer and / or tumors may be gastroesophageal cancers and / or tumors, for example, esophageal cancer, carcinoma, or tumors that are associated with lymph nodes or have metastasized to, for example, lymph nodes, liver, lungs, bones, or head and neck.

[0059] Therefore, it may be gastroesophageal cancer and / or tumor, for example, and / or esophageal cancer and / or tumor, carcinoma, or tumor may express the MAGE protein, peptide, antigen, or its peptide antigen, optionally as described herein, the MAGE-A4 protein, peptide, antigen, or its peptide antigen. According to the present invention, cancer and / or tumor may be recurrent or metastatic HNSCC having disease progression after platinum-containing chemotherapy and optionally expressing the MAGE-A4 protein, peptide, antigen, or its peptide antigen (e.g., the MAGE A4 peptide antigen, including GVYDGREHTV, SEQ ID NO: 2).

[0060] If esophagogastric junction cancer is lower esophageal junction cancer, it has a high probability of spreading to the stomach. Therefore, according to the present invention, cancer can be gastroesophageal cancer and / or tumor, e.g., stomach or gastric cancer, carcinoma, or tumor, which can be primary, secondary, recurrent, metastatic, or progressive.

[0061] Preferably, stomach or gastric cancer, carcinoma, or tumor may be selected from any one of the following: gastric cancer, gastric adenocarcinoma, gastric lymphoma, extranodal marginal zone B-cell lymphoma (MALT type), diffuse large B-cell lymphoma, gastric mesenchymal tumor, or hereditary diffuse gastric cancer.

[0062] Therefore, cancer may be gastroesophageal cancer and / or tumors, such as stomach or gastric cancer, carcinoma, or tumor, which may be associated with lymph nodes or have metastasized to, for example, the liver, lungs, bones, abdominal endometrium, and lymph nodes.

[0063] Therefore, stomach or gastric cancer, carcinoma, or tumor may express MAGE protein, peptide, antigen, or its peptide antigen, or optionally the MAGE-A4 protein, peptide, antigen, or its peptide antigen as described herein. According to the present invention, cancer may be recurrent or metastatic HNSCC having disease progression after platinum-containing chemotherapy and optionally expressing MAGE-A4 protein, peptide, antigen, or its peptide antigen as described herein (e.g., MAGE A4 peptide antigen including GVYDGREHTV, SEQ ID NO: 2).

[0064] Standard care Gastric and esophageal cancer and / or tumors include cancer and / or tumors of the esophagus, gastroesophageal junction, or stomach, i.e., gastric cancer.

[0065] Standard care for gastroesophageal cancer and / or tumors, including, optionally, gastroesophageal junction (EGJ) cancer and / or tumors, may be systemic platinum-based chemotherapy, consisting of cisplatin or carboplatin chemotherapy, or alternatively, any combination of cisplatin, fluorouracil, and leucovorin, or cisplatin and etoposide, or carboplatin and paclitaxel, or cisplatin and capecitabine, or alternatively, platinum-fluoropyrimidine doublets and, optionally, anthracyclines. Alternatively, standard care may be selected from treatment with any one of the following: ECF (epirubicin, cisplatin, 5-FU), ECX (epirubicin, cisplatin, capecitabine), EOF (epirubicin, oxaliplatin, 5-FU), and EOX (epirubicin, oxaliplatin, capecitabine).

[0066] Alternatively, standard care may be selected from treatment with lapatinib, FLOT / FOLFIRI chemotherapy, 5-fluorouracil, epirubicin, etoposide, docetaxel, oxaliplatin, capecitabine or irinotecan, ifosfamide, mytomycin C, vindesine, vinblastine, etoposide, gemcitabine, or one of the following taxanes such as paclitaxel (Taxol) or docetaxel. Alternatively, standard care may be selected from targeted therapies using cetuximab in combination with any one of the following: a human epidermal growth factor receptor 2 (HER2) inhibitor such as trastuzumab, lapatinib, or pertuzumab; a vascular endothelial growth factor (VEGF) inhibitor such as ramucirumab or bevacizumab; an EGFR inhibitor antibody such as panitumumab or cetuximab; a human hepatocyte growth factor (HGF) and / or Met inhibitor such as onarutuzumab or rilotumumab; a PD1 or PD-L1 inhibitor such as nivolumab, pembrolizumab, avelumab, atezolizumab, or cetuximab; or a taxane such as fluorouracil, methotrexate, cisplatin, carboplatin, paclitaxel (Taxol), or docetaxel.

[0067] Accordingly, the present invention, as well as the methods, treatments, and uses thereof, provide a reduction in MAGE-A4 expression or concentration in a subject compared to placebo treatment, or compared to no treatment, or compared to pre-treatment, or compared to treatment including standard care.

[0068] Disease biomarkers The present invention, as well as the methods, treatments, and uses and / or kits of the present invention, provide treatment, prevention, or delay of progression of gastroesophageal cancer and / or tumors in a subject, as determined by changes in the expression or concentration of a target disease biomarker compared to the expression or concentration of the disease biomarker before treatment, compared to placebo treatment or no treatment, or compared to treatment including standard care.

[0069] Changes in disease biomarker levels from baseline (before treatment) correlate with the response to treatment and correspond to the efficacy and success of cancer and / or tumor treatment.

[0070] Disease biomarkers for gastroesophageal cancer may include one or more of the following: glycan antigen (CA)72-4, CA19-9 glycolipid antigen, alpha-fetoprotein, glycan antigen (CA)12-5, SLE, BCA-225, hCG, pepsinogen I / II, carcinoembryonic antigen (CEA), HER2 proto-oncogene, CD44, or CA19-9, and these are biomarkers frequently used in clinical practice for gastroesophageal cancer and / or tumors.

[0071] Disease biomarkers for gastroesophageal cancer may also include expression or plasma levels of VEGF-A or angiopoietin-2 (Ang-2), overexpression or amplification of any of the following: EGFR, mesenchymal epithelial transition factor receptor (MET), hepatocyte growth factor (HGF), FGFR1, FGFR2, FGFR3, and FGFR4 (fibroblast growth factor receptors), FGFR amplification which induces receptor overexpression, chromosomal translocation, and point mutations, or enhanced kinase activity, expression levels of PD-1 and PD-L1, elevated levels of programmed death ligands 1 and 2 (PD-L1 / 2), and any one of the following gene mutations in known cancer-associated genes in gastroesophageal cancer and / or tumors, such as TP53, PTEN, ARID1A, APC, CTNNB1, CDH1, PI3KCA, and KMT2C.

[0072] Disease biomarkers for gastroesophageal cancer and / or tumors may also be selected from one or more of the following genes (tumor series): TP53, ELMO1, DOCK2, CDKN2A, ARID1A, SMAD4, PIK3CA, KRAS, HER2, EGFR, CND1, and MET, which may include gene mutations, deletions, or copy number alterations.

[0073] EBV-positive subtype gastroesophageal cancer and / or tumor biomarkers may include any of CXXC4, TIMP2, and PLXND1. COL9A2, EYA1, and ZNF365 are highly methylated in EBV-positive individuals, somatic mutations in BMP (bone morphogenetic protein), amplification or mutations in any of JAK2, MET, ERBB2, or PIK3CA, or mutations in ARID1A or BCO.

[0074] Disease biomarkers for gastroesophageal cancer and / or tumors include CXC chemokine receptor 2 (CXCR2) expression or mRNA expression, CC chemokine receptor 4 (CCR4) expression or mRNA expression, CC chemokine receptor 7 (CCR7) expression or mRNA expression, detected loss of heterozygosity in tumor cell-derived DNA, presence or level of hypermethylation of cytosine-phosphate-guanine (CpG) rich promoter regions, and metalloproteinase expression, such as MMP-1 or gelatinase MMP One or more of the following may be selected: -2 or MMP-9 or stromelysin, MMP-3 and MMP-10, interleukin IL-6 and IL-8 levels or expression, MAGE-A1, 2, 3, 4, 5, 6, cytokeratin (e.g., CK6, 16, or 17) expression, or actin or myosin concentration or expression level, overexpression of eukaryotic translation factor 4E (eIF4E), or mutation levels in DNA repair genes, such as those in the nucleotide excision repair (NER) group.

[0075] Disease biomarkers for gastroesophageal cancer and / or tumors may be further selected from one or more of the following: presence or level of anaplastic lymphoma kinase (ALK) translocation, presence or level of epidermal growth factor receptor (EGFR) mutation, presence or level of Kirsten rat sarcoma virus oncogene homolog (KRAS) mutation, human epidermal growth factor receptor-2 (HER2 / neu) expression, presence or level of B-Raf oncogene serine / threonine kinase (BRAF) mutation, presence or level of C-KIT oncogene mutation, expression of MAGE-A1, 2, 3, 4, 5, 6, and presence or level of Janus kinase 2 (JAK2) mutation.

[0076] Further disease biomarkers for gastroesophageal cancer and / or tumors may include any one of the following: EML4-ALK tyrosine kinase fusion, alterations in DNA methylation, epigenetic changes such as histone tail modifications, or microRNA regulation that leads to inactivation of tumor suppressor, as well as mutations and amplifications of c-MET, NKX2-1, LKB1, PIK3CA, and BRAF.

[0077] Disease biomarkers for gastroesophageal cancer and / or tumors may also include circulating tumor cells (CTCs), cell-free DNA, microRNA, cell-free RNA, and cell-derived vesicles such as exosomes that can circulate in the biological samples described herein. CTCs are disseminated tumor cells circulating in the bloodstream, and their presence is clinically associated with cancer or progressive or metastatic disease.

[0078] According to the present invention, disease biomarkers can be measured in, for example, biological samples of the subject described herein.

[0079] Biological samples Biological samples may be any subject or patient fluids that may contain disease biomarkers or cells or genetic material derived from cancer or tumors (e.g., gastroesophageal cancer and / or tumors, i.e., cancer and / or tumors of the esophagus, gastroesophageal junction, or stomach, i.e., gastric cancer), such as blood, serum, plasma, urine, tissue, cells, cell cultures, saliva, sputum, cerebrospinal fluid, lavage or fluids derived from the lungs, nasal cavity, bronchi, bronchoalveolars, esophagogastric tract or gastric tract, gastric or esophagogastric lavage or gastric juice, or peripheral blood samples, containing cell-derived vesicles such as circulating tumor cells (CTCs), cell-free DNA, microRNA, cell-free RNA, and exosomes. CTCs are disseminated tumor cells circulating in the bloodstream, either as single cells or as less numerous cell clusters, originating from either a primary tumor or metastasis. The presence of CTCs, like other disease biomarkers, is clinically associated with tumors and / or cancer, progressive or metastatic disease. For example, in gastroesophageal cancer and / or tumors, disease biomarkers or biomarkers or antibodies against MAGE-A4 may be detected as biomarkers for MAGE-A4-expressing cancer and / or tumors and / or tissues in biological samples, such as body fluids, such as fluids derived from the esophagus, gastric tract, or digestive tract, such as stomach or gastric lavage fluid or gastric juice.

[0080] Therapeutic effect Analysis of serum cytokines and soluble factors, and T cell infiltration of tumors The present invention, as well as the methods, treatments, and uses thereof, provide an increase in serum cytokine and / or interferon levels or concentrations in a subject compared to pre-treatment serum cytokine and / or interferon levels or concentrations, or compared to treatments including placebo, no treatment, or standard care.

[0081] Accordingly, the present invention, as well as the methods, treatments, and uses of the present invention, provide an enhancement or enhancement of cancer and / or tumor immunogenicity, measured, for example by the ability to induce an immune response in response to cancer and / or tumors or cancer and / or tumor antigens, as determined by increased secretion of cytokines and / or interferons, increased T cell proliferation, antigen responsiveness, target cell killing, T cell activation, CD28 signaling, tumor T cell infiltration, and increased ability to recognize and bind antigens presented by dendritic cells, compared to such levels before treatment or intervention, or compared to placebo, or compared to no treatment, or compared to treatment including standard care, by, for example, at least 10%, alternatively 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 120%, 150%, 200%, or more.

[0082] The efficacy of cancer immunotherapy is conditioned by the infiltration of tumors by activated tumor-specific T cells. The activity of these T cells, in turn, will be affected by the presence of an immunosuppressive environment (e.g., regulatory T cells) in the tumor. Therefore, direct assessment of the "immune landscape" inside the tumor is of great value in monitoring the efficacy of T cell immunotherapy and can be quantified by tumor biopsies that evaluate the immune state of the tumor before and after T cell injection. Thus, the present invention provides an improvement in T cell infiltration of the tumor and / or a decrease in T cell suppressor factors, determined by a decrease in the level of, for example, T-reg, myeloid-derived suppressor cells (MDSC), PD-L1 protein expression, CCL3, IL8, IL1β, CXCL10, or serum cytokine levels selected from sIL2Rα, or the level of inhibitory receptors selected from PD-1, CTLA-4, TIM-3, LAG-3, BTLA, or TIGIT, as compared to before treatment or without treatment, or as compared to treatment including standard care. Alternatively, an increase in the level of interferon-γ, interleukin-6, interleukin-10, cytokine production, such as IL-2, TNF-α, IFN-γ, and granzyme B, or innate immune cells such as NK cells, adaptive immune cells (CD4 + and CD8 + ), or an improvement in proliferation in T cells determined by, for example, the Ki67 expression level, as compared to before treatment or without treatment, or as compared to treatment including standard care described hereinbefore.

[0083] Tumor size and tumor burden The present invention, as well as the methods, treatments, and uses thereof, provide an improvement or enhancement of the level or response to reducing tumor growth or tumor growth rate, or maintaining tumor size after discontinuation of treatment, or to the level or response, as determined by, for example, measurement of tumor size or tumor number, compared to pre-treatment, placebo treatment, no treatment, or treatment including standard care, preferably an improvement or enhancement of at least 10%, alternatively 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 120%, 150%, 200%, or more, compared to pre-treatment, placebo treatment, no treatment, or treatment including standard care. Preferably, the improvement or enhancement of the level or response may be a continuous improvement or enhancement of the level or response and / or may have a duration of at least the same duration as the duration of treatment, at least 1.5, 2.0, 2.5, or 3.0 times the length of the duration of treatment, or more. Such an increase or enhancement of levels or response may be determined from RECIST 1.1 measurements [EA Eisenhauer., et.al., EUROPEAN JOURNAL OF CANCER 45 (2009) 228-247], or from tumor biopsy or liquid biopsy (peripheral blood-derived plasma) determining tumor-associated circulating free DNA (cfDNA) or exosomes (a stable source of mRNA). Both tumor burden and immune response can be monitored using exosomes (produced by all cells, including tumor cells and immune cells) and cfDNA (produced by dying tumor cells). Analysis of exosomes and cfDNA may enable (a) estimation of overall tumor burden and genetic profiling (including MAGE-A4 mRNA expression or mutation profiling) from exosomes and cfDNA, and (b) systemic assessment of immune response (gene expression by cytotoxic and regulatory immune cells) from exosomes.

[0084] As stated above, standard care treatment for gastroesophageal cancer and / or tumors may be as described above herein.

[0085] MAGE-A4 TCR+ Cell Persistence The present invention, as well as the methods, treatments, and uses thereof, provide improved therapeutic efficacy in gastroesophageal cancer and / or tumors in subjects, and improved treatment, inhibition, or delay of their progression, as determined by measuring the persistence of injected engineered and modified immune-responsive cells expressing or presenting heterologous T cell receptors (TCRs), as described herein, for example. The persistence of injected engineered and modified immune-responsive cells correlates with therapeutic efficacy and is also a measure of long-term safety. Cell persistence can be determined by qPCR or flow cytometry (FCM). For example, quantification of MAGE-A4 or MAGE-A4+CD8 TCR+ cells by PCR of transgenes from DNA extracted from frozen subject PBMCs can be used as an evaluation measure, similar to quantification of MAGE-A4 or MAGE-A4+CD8 TCR-expressing cells by FCM from frozen subject PBMCs. T cell phenotype and activity can be assessed using various assays, e.g., • Phenotypic analysis for T cell lineage determination in cell products before and after injection, and in the target blood. • Quantification of the senescence and activation status of T cells derived from target PBMCs. • Quantification of soluble factors reflecting the in vivo function of injected T cells, such as MAGE-A4 or MAGE-A4+CD8 TCR+ T cells. • Ex vivo activity of target transdextrins at various time points to assess the potential functionality of those cells before and / or during the treatment timeframe. It can be determined by [the relevant factor].

[0086] T cell function The present invention, as well as the methods, treatments, and uses thereof, provide enhancement of T cell function compared to pre-treatment, placebo-based treatment, no treatment, or treatment including standard care. Preferably, T cell function is enhanced by at least 10%, alternatively 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 120%, 150%, 200%, or more, as determined by, for example, increased secretion of γ-interferon from CD8+ T cells, increased T cell proliferation, increased internal signaling, increased antigen responsiveness, increased secretion of cytokines and / or interferon, increased target cell killing, increased T cell activation, increased CD28 signaling, increased T cell infiltration ability, and increased ability to recognize and bind antigens presented by dendritic cells.

[0087] According to the present invention, and the methods and uses thereof, tumor immunity or evasion of immune recognition by tumors can be weakened, resulting in improved tumor recognition and attack by the immune system, thereby treating tumor immunity as measured, for example, by tumor binding, tumor reduction, and tumor clearance. Accordingly, the present invention provides a treatment for tumor immunity and / or a treatment for tumor immunity that is enhanced by at least 10%, alternatively 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 120%, 150%, 200%, or more, compared to pre-treatment, placebo treatment, no treatment, or treatment including standard care, as measured, for example, by tumor binding, tumor reduction, or tumor clearance.

[0088] In the context of T cell activity, the term “dysfunction” refers to a state of reduced immune responsiveness to antigen stimulation, including T cell exhaustion and / or anergy, in which T cells are able to recognize and bind to antigens, such as cancer and / or tumor antigens or their peptide antigens, but exhibit reduced effectiveness in driving the immune response or fighting cancer progression and / or tumor growth. Dysfunctioning T cells exhibit an impaired ability to convert antigen recognition into downstream T cell effector functions such as proliferation, cytokine and interferon production, or target cell killing, as is characteristic of T cell dysfunction, as well as apparent unresponsiveness or unreliability to antigen recognition. “T cell dysfunction” may be associated with or detected as: insufficient increase in T cell signaling via PD-1; T cells having reduced ability to proliferate and / or produce cytokines and / or cytolytic activity; T cell anergy; and tumor immunity.

[0089] "T cell exhaustion" includes a state of T cell dysfunction resulting from continuous TCR signaling as part of the response to cancer, which inhibits the optimal response to the tumor. Exhaustion can be exerted through intracellular negative regulatory (co-stimulatory) pathways (e.g., PD-1, PD-1 axis, B7-H3, B7-H4) or through extracellular negative regulatory pathways (immunomodulatory cytokines). T cell exhaustion is characterized by poor effector function, continuous expression of inhibitory receptors, and altered transcriptional activity distinct from that of functional effector or memory T cells. T cell anergy arises through deficient signaling via T cell receptors and, more often in co-stimulatory situations, a resulting state of unresponsiveness to antigenic stimulation, as such T cells do not undergo clonal growth and / or acquire effector function.

[0090] Treatment and administration According to the present invention, modified immune-responsive cells can be administered sequentially or intermittently, either as a single dose or in more than one dose, at the discretion of the present invention.

[0091] Therefore, modified immune-responsive cells can be administered as a single dose or in multiple doses. Modified immune-responsive cells can be administered in doses ranging from approximately 500 million cells to any one of the following: approximately 1 billion cells, 2 billion cells, 3 billion cells, 4 billion cells, 5 billion cells, 6 billion cells, 7 billion cells, 8 billion cells, 9 billion cells, 10 billion cells, 11 billion cells, 12 billion cells, 13 billion cells, 14 billion cells, 15 billion cells, 16 billion cells, 17 billion cells, 18 billion cells, 19 billion cells, 20 billion cells, or 21 billion cells. Modified immune-responsive cells can be administered in doses of approximately 100 million to 200 million cells, 300 million to 400 million cells, 500 million to 600 million cells, 700 million to 800 million cells, or 900 million to 1 billion cells, or optionally 500 million to 1 billion cells, 2 billion to 5 billion cells, or 6 billion to 10 billion cells.

[0092] According to the present invention, modified immune-responsive cells can be administered intravenously, intramuscularly, subcutaneously, locally, orally, percutaneously, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intravenously, or intranasally, or by intravenous injection. Preferably, modified immune-responsive cells can be administered intravenously or by intravenous injection.

[0093] According to the present invention, modified immune-responsive cells are (a) Single dose in each of one or more medication cycles, (b) One or more doses in each of one or more medication cycles, (c) Single dose on day 1 of each of one or more medication cycles, (d) One or more doses in each of one or more medication cycles, including the dose on day 1 of each of one or more medication cycles. (e) One or more doses in each of one or more medication cycles, where at least one dose is on day 1 of each cycle. (f) Single dose It may be administered as such.

[0094] According to the present invention, modified immune-responsive cells may be administered in a drug delivery cycle, which may be any of 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28 weeks, or any of 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months (for example, since the previous last drug delivery). Therefore, a drug delivery cycle may be any of 10-12 weeks, 11-13 weeks, 14-17 weeks, 18-21 weeks, 22-24 weeks, 24-27 weeks, 28-30 weeks, 3 months, 4 months, 5 months, or 6 months (for example, since the previous last drug delivery).

[0095] According to the present invention, modified immune-responsive cells may be administered in a drug delivery cycle, and the drug delivery cycle is (a) Disease progression following previous administration of modified immune-responsive cells, and / or (b) at least 12 weeks after previous administration of modified immune-responsive cells It may be in, or may begin from, or may resume from, (c) Tumors and / or cancers express MAGE-A4 and / or its peptide antigen, and / or (d) MAGE-A4 and / or its peptide antigen are detected in the biological sample and / or exceed the normal range.

[0096] According to the present invention, modified immune-responsive cells may be administered in a drug delivery cycle, and the drug delivery cycle is (a) a confirmed response, complete response, or partial response following prior administration of modified immune-responsive cells, or (b) stable disease for a period of two, three, or four months or longer, or equivalent, following prior administration of modified immune-responsive cells, followed by disease progression, and / or (c) After more than 12 weeks or equal to the previous administration of modified immune-responsive cells It may be in, or may begin from, or may resume from, (d) Tumors and / or cancers express MAGE-A4 and / or its peptide antigen, and / or (e) MAGE-A4 and / or its peptide antigen are detected in the biological sample and / or exceed the normal range.

[0097] Tumors and / or cancers may express MAGE-A4 and / or its peptide antigen at levels exceeding or equal to an immunohistochemical intensity of 1+ as determined by immunohistochemistry, and / or at immunohistochemical antigen expression frequencies exceeding or equal to 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50%, preferably exceeding or equal to 30 or 32%. The MAGE-A4 and / or peptide antigen levels of the target biological sample exceeding the normal range may be above or equal to 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, or 500 ng / mL, preferably above or equal to 50 or 100 ng / mL.

[0098] According to the present invention, the dose may be a fixed dose or a variable dose. For example, if more than one dose is administered, i.e., in the case of multiple doses, the dose may be fixed or variable, and if more than one dose is administered, the dose may be increased or increased, for example, in each dosing cycle, i.e., increasing levels of dose, for example, gradually, for example, from 100 million cells to 500 million cells to 1 billion cells to 5 billion cells to 10 billion cells.

[0099] According to the present invention, modified immune-responsive cells are preferably administered as a single dose of approximately 5 billion to 10 billion cells.

[0100] According to the present invention, modified immune-responsive cells may be administered for a specified period, meaning that a modified immune-responsive cell administration cycle may be performed for a specified period. The specified period may be any of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, or 48 months, preferably 24 months.

[0101] According to the present invention, the method is (a) Modified immune-responsive cells are administered as a single dose, (b) If the disease state is determined during the period following the administration of modified immune-responsive cells and compared to the state before the administration of modified immune-responsive cells, and a progressive disease is determined, (c) Modified immune-responsive cells are administered as a single dose, and optionally, tumors and / or cancers express MAGE-A4 and / or its peptide antigen, and / or MAGE-A4 and / or its peptide antigen is detected in the biological sample and / or exceeds the normal range. The steps may include. Preferably, the period is greater than or equal to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48 weeks, preferably greater than or equal to 12 weeks.

[0102] According to the present invention, the method is (a) Modified immune-responsive cells are administered as a single dose, (b) If the disease state is determined in the first and second periods after administration of modified immune-responsive cells and compared to the state before administration of modified immune-responsive cells, and a stable disease is determined after the first period and a progressive disease is determined after the second period, (c) Modified immune-responsive cells are administered as a single dose, and optionally, tumors and / or cancers express MAGE-A4 and / or its peptide antigen, and / or MAGE-A4 and / or its peptide antigen is detected in the biological sample and / or exceeds the normal range. The steps may include: Preferably, the first period is greater than or equal to one of 1, 2, 3, 4, 5, 6, 7, or 8 months, preferably greater than or equal to 4 months.

[0103] Preferably, the second period is greater than or equal to any one of the following periods after the first period: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, or 48 months, preferably greater than or equal to 4 months.

[0104] According to the present invention, "complete response" (CR) is determined when all target lesions or tumors are assessed or measured as having disappeared. "Partial response" (PR) is determined when there is a measurement of at least a 30% reduction in the sum of the longest diameters (SLD) of the target lesions or tumors, for example with reference to a control or pre-treatment comparator. "Progressive disease" (PD) is determined when there is a measurement of at least a 20% increase in the sum of the longest diameters (SLD) of the target lesions or tumors, or the presence of one or more new lesions, since the initiation of treatment, for example with reference to a control or pre-treatment comparator. "Stable disease" (SD) is determined when, using the smallest SLD since the initiation of treatment as a reference, there is neither a sufficient reduction or decrease in the sum of the longest diameters (SLD) of the target lesions or tumors to the quality of PR nor a sufficient increase to the quality of PD.

[0105] According to the present invention, the pre-treatment subject is tumor and / or cancer cells, as determined by immunohistochemistry, with tumor and / or cancer cell MAGE-A4 and / or its peptide antigen expression exceeding or equal to an intensity of 1+ in immunohistochemistry, and / or 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, The immunohistochemical antigen expression frequencies may include 43, 44, 45, 46, 47, 48, 49, or more than or equal to 50%, preferably more than or equal to 30 or 32%, and the non-cancerous MAGE-A4 and / or its peptide antigen expression is less than or equal to 1, 2, 3, 5, 6, 7, 8, 9, or 10% of cells in non-cancerous or non-tumor tissue at any intensity by immunohistochemistry, preferably less than or equal to 1% or 5%.

[0106] According to the present invention, the pretreatment subject may contain biological sample levels of MAGE-A4 and / or its peptide antigen greater than or equal to 10, 25, 50, 100, 200, 300, or 400 ng / mL, preferably greater than or equal to 50 ng / mL, wherein MAGE-A4 expression is less than or equal to 1, 2, 3, 5, 7, 9% or less than 10%, preferably less than 1 or 5%, of cells in noncancerous or nontumor tissue at any intensity determined by immunohistochemistry.

[0107] According to the present invention, pre-treatment subjects may include 0-1 Eastern Cooperative Oncology Group (ECOG) and / or measurable diseases (gastroesophageal cancer and / or tumors) according to the Response Evaluation Criteria in Solid Tumors (RECIST) 1.1, and / or histologically confirmed gastroesophageal cancer and / or tumors.

[0108] According to the present invention, the subject before treatment is determined to be HLA-A*02 positive, and / or the subject's cancer or tumor preferably exhibits expression of MAGE-A4 and / or its peptide antigen, such as MAGE-A4 RNA or protein, as described above herein.

[0109] According to the present invention, before treatment, the subject is (a) The HLA-A genotype is HLA-A*02:07P as the sole HLA-A*02 allele. (b) The HLA-A genotype is HLA-A* of any A*02 null allele as the sole HLA-A*02 allele, or (c) Symptomatic CNS metastasis If a patient has one or more of the following conditions, they are not eligible for treatment.

[0110] According to the present invention, subjects may be positive for HLA-A*02 selected from, for example, HLA-A*02:01, HLA-A*02:02, HLA-A*02:03, HLA-A*02:04, HLA-A*02:05, HLA-A*02:06, HLA-A*02:642, or HLA-A*02:07, preferably HLA-A*02:01 or HLA-A*02:642, and / or gastroesophageal cancer and / or tumors express MAGE-A4, MAGE-A4 peptide antigen, GVYDGREHTV, SEQ ID NO: 2, and MAGE-A4 peptide antigen.

[0111] According to the present invention, the subject may preferably be intolerant to standard care treatment as described herein, and additionally or alternatively, the subject and / or cancer and / or tumor may have previously failed treatment with, for example, standard care treatment as described herein, or treatment with any of the following: surgery (resection), radiotherapy, targeted therapy, immunotherapy, or chemotherapy, or chemotherapy in combination with surgery (resection), radiotherapy, radiotherapy, targeted therapy, or immunotherapy; or treatment with local area therapy, optionally selected from chemical and / or thermal percutaneous ablation and intra-arterial chemoembolization (chemoembolization).

[0112] According to the present invention, cancers that are gastroesophageal cancer and / or tumors may be primary cancers, secondary cancers, relapsing or refractory cancers or recurrent or locally recurrent cancers, advanced or locally advanced or metastatic cancers, unresectable or locally limited cancers for which there are no surgical or radiotherapy options, or inoperable cancers, cancers unsuitable for transplantation or localized therapy, or any combination thereof. The subjects may have relapsing or refractory cancers or recurrent or locally recurrent cancers or metastatic cancers or locally limited or inoperable cancers, or any combination thereof.

[0113] Preferably, gastroesophageal cancer and / or tumor is inoperable and / or metastatic and / or advanced and / or locally advanced gastroesophageal cancer and / or tumor, for example, any of the esophageal, gastroesophageal junction cancer and / or tumor, or gastric cancer and / or tumor, preferably esophageal squamous cell carcinoma (ESCC), esophageal adenocarcinoma (EAC), or esophagogastric junction cancer, carcinoma, adenocarcinoma, or tumor (EGJ), or stomach or gastric cancer, carcinoma, or tumor, preferably inoperable, metastatic, advanced, or locally advanced esophagogastric junction cancer, carcinoma, adenocarcinoma, or tumor (EGJ).

[0114] Further, the present invention provides treatments or uses in which a subject has not received prior treatment for gastroesophageal cancer and / or tumors, or alternatively, a subject has received prior treatment for gastroesophageal cancer and / or tumors and / or has not responded to prior treatment.

[0115] According to the present invention, prior treatment may include one or more systemic and / or local therapies, such as surgery, radiotherapy, cryotherapy, laser therapy, local therapy, chemotherapy, hormone therapy, targeted drugs, or immunotherapy. Therefore, prior treatment may include one or more local therapies, such as surgery, radiotherapy, cryotherapy, laser therapy, or local therapy, and / or one or more systemic therapies, such as chemotherapy, hormone therapy, targeted drugs, or immunotherapy. According to the present invention, prior treatment may include one of the following: systemic therapy for the initial diagnosis of a localized disease, systemic therapy after the diagnosis of a recurrent or metastatic disease, systemic therapy after an oligometastatic disease, or systemic therapy after a locally recurrent disease.

[0116] Therefore, if the treatment is for gastroesophageal cancer and / or tumors, prior treatment may include the relevant standard care described herein for the treatment of primary tumors in locally advanced or metastatic settings, for example, failed due to advanced disease or unacceptable toxicity or intolerance. Therefore, if the treatment is for gastroesophageal cancer and / or tumors, prior treatment may include systemic platinum-containing chemotherapy, which may be selected from cisplatin or carboplatin chemotherapy, for the treatment of primary tumors in locally advanced or metastatic settings, for example, failed due to advanced disease or unacceptable toxicity or intolerance. Therefore, if the treatment is for gastroesophageal cancer and / or tumors, prior treatment may include cisplatin, fluorouracil, and leucovorin, or cisplatin and etoposide, or carboplatin and paclitaxel, or a combination of cisplatin and capecitabine, alternatively platinum-fluoropyrimidine doublets and optionally anthracyclines; optionally, immune checkpoint inhibitors such as pembrolizumab or nivolumab during or after platinum-based chemotherapy; any of the following may be included: targeted therapies or targeted antibody therapies such as trastuzumab, cetuximab, bevacizumab, or ramucirumab; optionally, prior treatment has failed, for example, due to advanced disease or unacceptable toxicity or intolerance.

[0117] According to the present invention, the prior treatment may include a PD-1 axis-binding antagonist, a PD-L1-binding antagonist, or a PD-1-binding antagonist. Therefore, the prior treatment is (a) Anti-PD-L1 antibodies that inhibit the binding between PD-L1 and PD-1 and / or between PD-L1 and B7-1, (b) Anti-PD-L1 antibodies that inhibit PD-L1 on the surface of cancer cells from converting signals into intracellular pathways, (c) Anti-PD-1 antibodies that inhibit the binding between PD-L1 and PD-1 and / or between PD-L2 and PD-1. (d) Anti-PD-1 antibodies that inhibit PD-1 on the surface of T cells from converting signals into intracellular pathways, (e) (i) durvalumab, Imfinzi, or MEDI4736, (ii) Atezolizumab, Tecentriq, or MPDL3280A, (iii) Avelumab, Bavencio, or MSB0010718C (iv) MDX-1105, BMS-936559 A PD-L1-binding antagonist selected from the following, (f) (i) Pembrolizumab, Keytruda, lambrolizumab, or MK-3475, (ii) Semiprimab, Libtayo, or REGN-2810, (iii) BMS / ONO, nivolumab, Opdivo, ONO-4538, BMS-936558, or MDX1106 Selected PD-1 binding antagonist It may include.

[0118] According to the present invention, prior treatment may include an epidermal growth factor receptor antagonist, optionally cetuximab. According to the present invention, if prior treatment includes chemotherapy, it may optionally include one or more platinum compounds selected from lipoplatin, cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenantriplatin, satraplatin, and picoplatin. Additionally or alternatively, if prior treatment includes chemotherapy, it may include one or more chemotherapeutic agents selected from methotrexate, capecitabine, taxanes, anthracyclines, paclitaxel, docetaxel, paclitaxel protein-binding particles, doxorubicin, epirubicin, 5-fluorouracil, cyclophosphamide, afatinib, vincristine, etoposide, or a combination thereof. Additionally or alternatively, if prior treatment includes chemotherapy, this may include one or more chemotherapeutic agents selected from FEC: 5-fluorouracil, epirubicin, cyclophosphamide; FAC: 5-fluorouracil, doxorubicin, cyclophosphamide; AC: doxorubicin, cyclophosphamide; EC: epirubicin, cyclophosphamide. According to the present invention, prior treatment may include one or more of sorafenib, PD1 or PD-L1 antagonists or inhibitors, regorafenib, cabozantinib, sunitinib, brivanib, everolimus, tivantinib, linifanib, or optionally one or more local therapies selected from chemical and / or thermal percutaneous ablation and intra-arterial chemoembolization, optionally, prior treatment has failed, for example, due to progressive disease or unacceptable toxicity or intolerance.

[0119] According to the present invention, the subject may not have received prior treatment in a relapse occurring less than 12 months or equal to 12 months after the last treatment, or less than 6 months or equal to 12 months after the last treatment. According to the present invention, the subject may not have received any prior adjuvant therapy (surgery, followed by radiation and / or chemotherapy) in a relapse occurring less than 12 months or equal to 12 months after the last treatment, or less than 6 months or equal to 12 months after the last treatment.

[0120] According to the present invention, the treatment is compared to a control such as placebo treatment, or before treatment, or no treatment, or treatment including standard care as described herein. (a) Progression-free survival, (b) progression-free period, (c) Duration of response, (d) overall survival; (e) Objective response or objective response rate, (f) Total response or total response rate, (g) Partial response or partial response rate, (h) Complete response or complete response rate; (i) Rate of stable disease or median of stable disease (j) median progression-free survival, (k) median progression-free interval, (l) median duration of response, or (m) Median overall survival; (n) Median objective response or median objective response rate, (o) Median overall response rate or median overall response rate, (p) median partial response or median partial response rate, (q) Median complete response or median complete response rate, (r) Median rate of stable disease or median of stable disease, To extend or improve, or to effectively extend or effectively improve.

[0121] According to the present invention, the treatment is compared to a control such as placebo treatment, or before treatment, or no treatment, or treatment including standard care as described herein. (a) Best Overall Response (BOR), (b) Time to Confirmed Response (TTR), (c) Duration of Response (DoR), (d) Duration of Stable Disease (DoSD), (e) Progression-Free Survival (PFS), or (f) Overall Survival (OS) To extend or improve, or effectively extend or effectively improve, one or more of the following.

[0122] Best Overall Response (BOR) may be defined as the best response recorded from the day of T-cell infusion to disease progression. Time to Confirmed Response (TTR) may be defined as the period between T-cell infusion and the first day of confirmed response. Duration of Response (DoR) may be defined as the period from the first day of confirmed response to the day of disease progression, disease progression (or death). Duration of Stable Disease (DoSD) may be defined as the period from the day of T-cell infusion to the day of disease progression, disease progression (or death). Progression-Free Survival (PFS) may be defined as the interval between the day of T-cell infusion and the earliest date of disease progression or death due to any cause, according to RECIST v1.1. Overall Survival (OS) may be defined as the period between T-cell infusion and death due to any cause.

[0123] Progression-free survival (PFS) refers to the time from treatment (or randomization) to the first disease progression or death. Progression-free progression (TTP) is otherwise equivalent to PFS, but does not count patients who die from causes other than the treated cancer or tumor. Duration of response (DoR) is the length of time that cancer, tumor, or lesion continues to respond to treatment without growing or spreading. According to the present invention, DoR, TTP, and PFS may be assessed by the Response Evaluation Criteria in Solid Tumors (RECIST), or by CA-125 levels (cancer antigen 125) as a determinant of progression, or optionally by referring to the expression of disease biomarkers or MAGE-A4 in the subject biological sample, cancer, and / or tumor tissue or cells, i.e., MAGE-A4 protein, peptide, or mRNA. Duration of response, rate, readout, evaluation scale, or time point may be measured from the day treatment begins, e.g., the day modified immune-responsive cells are administered to the subject, or the day standard care or placebo is administered.

[0124] According to the present invention, PFS and / or TTP and / or DoR, or the median thereof, can be extended or improved by at least 1, 2, 3, or 4 weeks, 1 month, 2 months, 2.3 months, 2.5 months, 2.9 months, 3 months, 3.5 months, 3.8 months, 4 months, 4.5 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 16 months, 18 months, 20 months, 22 months, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, or 10 years compared to placebo treatment, or compared to pre-treatment, or compared to no treatment, or compared to treatment including standard care as described herein (control).

[0125] In one embodiment, PFS and / or TTP and / or DoR, or the median thereof, are extended by approximately 2.9 to 3.8 months compared to the control. In another embodiment, PFS and / or TTP and / or DoR, or the median thereof, are extended by at least approximately 3.8 months compared to the control. In yet another embodiment, PFS and / or TTP and / or DoR, or the median thereof, are extended by approximately 2.3 months compared to placebo treatment, or compared to pre-treatment, or compared to no treatment, or compared to treatment including standard care as described herein (control), and in one embodiment, PFS and / or TTP and / or DoR, or the median thereof, are extended by approximately 6 months.

[0126] "Overall survival" refers to the period of time a subject remains alive. According to the present invention, overall survival or its median is improved or extended by approximately 1 month, 2 months, 2.3 months, 2.5 months, 2.9 months, 3 months, 3.5 months, 3.8 months, 4 months, 4.5 months, 5 months, approximately 6 months, approximately 7 months, approximately 8 months, approximately 9 months, approximately 10 months, approximately 11 months, approximately 12 months, approximately 1.5 years, approximately 2 years, approximately 3 years, approximately 4 years, approximately 5 years, approximately 6 years, approximately 7 years, approximately 8 years, approximately 9 years, or approximately 10 years from the start of the method or treatment according to the present invention, or from the initial diagnosis, and the event used for survival analysis at any discretion may be death from any cause. "Survival" refers to the period of time a subject remains alive and includes progression-free survival (PFS) and overall survival (OS). "Overall survival" is the length of time from the date of diagnosis or the initiation of treatment for the disease, tumor, and / or cancer, during which a subject diagnosed with a disease is still alive. Survival may be estimated by the Kaplan-Meier method, and any difference in survival may be calculated using a stratified log-rank test; "extending survival" or "increasing the likelihood of survival" means increasing PFS and / or OS in a treated subject compared to placebo treatment, or compared to pre-treatment, or compared to no treatment, or compared to treatment including, for example, standard care as described herein. According to the present invention, overall survival or survival can be extended or improved by at least one of the following periods compared to placebo treatment, or compared to before treatment, or compared to no treatment, or compared to treatment including standard care (control): 1 month, 2 months, 2.3 months, 2.5 months, 2.9 months, 3 months, 3.5 months, 3.8 months, 4 months, 4.5 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 16 months, 18 months, 20 months, 22 months, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, or 10 years.

[0127] "Objective response rate" (ObRR) is the proportion of subjects having a predetermined amount of tumor size reduction, determined by the sum of the longest diameters (SLD) of target lesions or tumors and over the shortest period of time, at the discretion of the patient. "Overall response rate (ORR)" is defined as the proportion of subjects having a partial or complete response to therapy; stable disease is not included. ORR is generally defined as the sum of complete responses (CR) and partial responses (PR) over a specified period of time. According to the present invention, ObRR and / or ORR and / or PR and / or CR and / or SD can be extended or improved by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, and 95% compared to placebo treatment, or compared to pre-treatment, or compared to no treatment, or compared to treatment including standard care as described herein.

[0128] According to the present invention, the method may further include the step of determining the expression level of a biomarker in a sample derived from a subject, a biological sample, comparing the level of the biomarker to a reference level to determine the likelihood of the subject responding to treatment, or to the level of the subject responding to treatment, wherein the sample is acquired either before, during, or after treatment. The reference level may be the level of the subject before treatment, or a level associated with the presence or absence of cancer. The biomarker may be a T-effector-related gene, such as CD8A, perforin (PRF1), granzyme A (GZMA), granzyme B (GZMB), interferon-γ (IFN-v), CXCL9, or CXCL10. The biomarker may be an activated stroma-related gene, such as transforming growth factor-β (TGF-β), fibroblast-activating protein (FAP), podoplanin (PDPN), collagen gene, or biglycan (BGN). The biomarker may be a myelokJ-derived suppressor cell-related gene, such as CD68, CD163, FOXP3, or androgen regulatory gene 1. Alternatively, the biomarker may be PD-L1, CD8, or an androgen receptor (AR) gene. Alternatively, the biomarker may be a disease biomarker previously described herein.

[0129] According to the present invention, subjects undergo lymphatic depletion chemotherapy prior to the administration of modified immune-responsive cells expressing or presenting heterologous T cell receptors (TCRs). Lymphatic depletion chemotherapy may include the administration of cyclophosphamide and / or fludarabine. Preferably, cyclophosphamide is administered in doses of about 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, or 850 mg / m². 2 / d[d=day], preferably about 500 or 600 mg / m² 2The drug is administered in doses of / d, preferably for 1 day, 2 days (x2d), 3 days (x3d), 4 days (x4d), or 5 days (x5d). Preferably, fludarabine is administered in doses of about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, or 85 mg / m². 2 The drug is administered in doses of / d, preferably for 1 day, 2 days (x2d), 3 days (x3d), 4 days (x4d), or 5 days (x5d). Preferably, lymphatic depletion chemotherapy is optionally administered at 500 mg / m². 2 In doses of cyclophosphamide at 20 mg / m² / d x 3d and fludarabine at 20 mg / m² / d x 3d, or 600 mg / m² 2 Cyclophosphamide at 3 / d x 3d and 30 mg / m² 2 The treatment involves the administration of cyclophosphamide and fludarabine at a dose of fludarabine / d × 4d. According to the present invention, lymph depletion chemotherapy may be administered 3, 4, 5, 6, 7, 8, 9, or 10 days prior, preferably 7 to 5 or 7 to 4 days prior, to the administration of modified immune-responsive cells expressing or presenting xenotypic T cell receptors (TCRs). The administration of cyclophosphamide and fludarabine may be sequential, separate, or simultaneous, and the administration may be intravenous or by intravenous infusion.

[0130] The present invention relates to subjects with gastroesophageal cancer and / or tumors, and includes, optionally, a step of administering a therapeutic regimen to a subject comprising an effective amount of modified immune-responsive cells expressing or presenting a heterologous T cell receptor (TCR) that binds to MAGE-A4 or a peptide antigen of MAGE-A4, or GVYDGREHTV, SEQ ID NO: 2, compared to pre-treatment, placebo-based treatment, no treatment, or standard care as described herein, as described previously herein with reference to the method of treatment and aspects, embodiments, and characteristics relating thereto. (a) For example, reducing the expression or concentration of the target MAGE-A4 in the target biological sample, (b) Enhance immune function, (c) reducing tumor growth or tumor growth rate, maintaining tumor size after discontinuation of treatment, or reducing the number or volume of tumors. (d) Increase serum cytokine and / or interferon levels or concentrations (e) To improve T cell persistence, (f) To improve T cell infiltration of tumors (g) Inducing changes in disease biomarkers that indicate effective treatment for gastroesophageal cancer and / or tumors, Further methods will be provided.

[0131] Accordingly, the present invention relates to a method for enhancing immune function, comprising the step of administering a therapeutic regimen to a subject having gastroesophageal cancer and / or tumor, as previously described herein with reference to the method of treatment and its aspects, embodiments and characteristics, comprising, optionally, compared to pre-treatment or placebo treatment or no treatment or treatment including standard care as described herein, the therapeutic regimen comprising an effective amount of modified immune-responsive cells expressing or presenting a heterologous T cell receptor (TCR) that binds to MAGE-A4 or MAGE-A4 peptide antigen, or GVYDGREHTV, SEQ ID NO: 2, respectively. (a) CD8 T cells in the subject have enhanced priming, activation, proliferation, and / or cytolytic activity. (b) The number of CD8 T cells increases in the subjects. (c) Cancer and / or tumor cells in the subject selectively have elevated expression of MHC class I antigen, and PBMC cells in the subject are randomly selected not to have elevated expression of MHC class I antigen. (d) The antigen-presenting cells in the subject have enhanced maturation and activation, and the antigen-presenting cells are dendritic cells at random selection. (e) Serum levels of IL-10 and / or IL-8 in the subjects decrease. (f) The cancer and / or tumor in question has an elevated level of T cell infiltration, (g) The target T cells have a reduced level of T cell PD-1 expression. Provide a method.

[0132] Therefore, referring to the above and the treated subjects, (a) CD8 T cell activation is gamma-IFN + (b) Maturation of antigen-presenting cells may be characterized by an increase in the frequency of CD8 T cells and / or enhanced cytolytic activity; (b) Maturation of CD83 + (c) The activation of antigen-presenting cells may be characterized by an increased frequency of dendritic cells; (d) CD8 T cells may be antigen-specific CD8 T cells.

[0133] According to the present invention, (a) A kit comprising an effective amount of modified immune-responsive cells expressing or presenting a heterologous T cell receptor (TCR) that binds to MAGE-A4 or MAGE-A4 peptide antigen, or GVYDGREHTV, SEQ ID NO: 2, and a package insert containing instructions for use of the modified immune-responsive cells to treat or delay the progression of gastroesophageal cancer and / or tumors in a subject. (b) an effective amount of modified immune-responsive cells expressing or presenting a heterologous T cell receptor (TCR) that binds to MAGE-A4 or the MAGE-A4 peptide antigen, or GVYDGREHTV, SEQ ID NO: 2, and in subjects having gastroesophageal cancer and / or tumors as described herein prior to this date, (i) For example, reducing the expression or concentration of the target MAGE-A4 in the target biological sample, (ii) Enhance immune function, (iii) reducing tumor growth or tumor growth rate, maintaining tumor size after discontinuation of treatment, or reducing the number or volume of tumors. (iv) Increase serum cytokine and / or interferon levels or concentrations (v) Improve T cell persistence, (vi) to improve T cell infiltration of tumors, or (vii) Induce changes in disease biomarkers that indicate effective treatment for gastroesophageal cancer and / or tumors. A kit including an accompanying document with instructions for use of modified immune-responsive cells in the method. It will be provided.

[0134] The present invention will be further described by reference to the following figures and embodiments. [Examples]

[0135] [Example 1] A phase I, open-label clinical trial to evaluate the safety and antitumor activity of autologous T cells expressing MAGE-A4-specific enhanced TCRs, specifically MAGE-A4 CD8 TCRs, in HLA-A2+ subjects with MAGE-A4-positive esophagogastric junction (EGJ) cancer and gastric cancer.

[0136] method The following presents human studies on genetically modified ADP-A2M4CD8 SPEAR T cells in subjects with HLA-A*02 and MAGE-A4-positive, inoperable, locally advanced or metastatic tumors of the esophagogastric junction (EGJ) or gastric cancer.

[0137] The diseases were histologically or cytogenetically confirmed and / or measurable diseases, documented according to RECIST v1.1 criteria. Subjects who were eligible based on HLA type and met the MAGE-A4 criteria were screened for overall health status, performance status, and disease stage. After screening, subjects who met all eligibility criteria underwent leukocyte apheresis to obtain cells for the production of autologous MAGE-A4 CD8 TCR-carrying T cells. Eligible subjects had an ECOG performance status of 0-1 required before lymphoid depletion, adequate organ function, and measurable disease, and (a) The subjects are positive for at least one HLA-A*02 encapsulated allele. (b) The subjects have inoperable or metastatic (progressive) esophageal (squamous or adenocarcinoma), esophagogastric junction (EGJ), or gastric cancer. (c) The subjects may have previously received fluoropyrimidine (e.g., fluorouracil or capecitabine) and / or platinum regimens. (d) The cancer and / or tumor may have Her2neu amplification but has failed (advanced disease or unacceptable toxicity) or has rejected trastuzumab. (e) The patient may have received no more than three prior systemic regimens. (f) The subjects have a histologically or cytologically confirmed diagnosis of metastatic (progressive) esophageal (squamous or adenocarcinoma), esophagogastric junction (EGJ), or gastric cancer.

[0138] Exclusion of subjects is primarily based on HLA-A genotype, i.e., whether the subject is positive for any HLA-A*02 allele other than the internal allele HLA-A*02:07P or the HLA-A*02 null allele (both HLA-A*02:07P and HLA-A*02 null alleles are alleles with low activity; therefore, if these alleles are the only 02 allele of a subject, they will be ineligible). Exclusion of subjects additionally includes those with symptomatic CNS metastases, or active autoimmune or immune-mediated diseases, or infections.

[0139] Following leukocyte apheresis, cells were transduced with MAGE-A4 CD8 TCR T cells (SEQ ID NOs. 5, 7 + SEQ ID NO. 3) specific to the MAGE-A4 antigen (particularly the specific MAGE-A4 antigen peptide, SEQ ID NO. 2). The cells were then enlarged and cryopreserved for later use. As soon as the MAGE-A4 CD8 TCR T cells became available, subjects received lymphocyte depletion chemotherapy with cyclophosphamide + fludarabine on days -7 to -5 or -7 to -4, followed by infusion of the transduced cells on day 1.

[0140] Three target cohorts were treated with drug administration using 100 million to 5 billion transduced cells, without dose escalation: Dose per 100mn cells, (cyclophosphamide: 500mg / m²) 2 / d)×3d; (Fludarabine: 20mg / m²) 2 / d)×3d 1bn cell dose, (cyclophosphamide: 500 mg / m²) 2 / d)×3d; (Fludarabine: 20mg / m²) 2 / d)×3d 5bn cell dose, (cyclophosphamide: 600 mg / m²) 2 / d)×3d; (Fludarabine: 30mg / m²) 2 / d)×4d

[0141] Participants will be hospitalized for 7 days after infusion, and then monitored for safety, T cell persistence, and cytokine production with CT and MRI scans performed at weeks 4, 8, 16, and 24, and every 3 months, until disease progression or early withdrawal from intervention. Annual long-term follow-up is planned for a period of 15 years.

[0142] Participants will be considered to have completed the intervention phase of the study if they receive a T-cell infusion and subsequently experience disease progression or die before disease progression. Optionally, a second T-cell infusion may be administered, and they will remain in the intervention phase of the study until they experience further disease progression. Once progression is confirmed, no further efficacy assessments other than overall survival will be performed. All participants who complete the intervention phase of the study will enter a long-term follow-up (LTFU) phase for observation of delayed adverse events (AEs) for 15 years after infusion, in accordance with FDA and EMA regulations. This study will be considered complete when the last surviving participant completes the LTFU.

[0143] To assess the safety and tolerability of MAGE-A4 CD8 TCR T cells, the incidence of dose-limiting toxicity (DLT) is monitored, and the optimally tolerable dose range, adverse events (AEs), and severe adverse events (SAEs) are determined; laboratory assessments including chemistry, hematology, and coagulation; and cardiac assessments including ECG and cardiac troponin are performed.

[0144] During the study, MAGE-A4 was evaluated as a biomarker for tumor MAGE-A4 expression and antitumor activity. This was done to correlate the levels of antigen expression at the tumor level at baseline and after MAGE-A4 CD8 TCR T cell infusion. Time-course MAGE-A4 expression in tumors after therapy was assessed to determine tumor immunity or resistance to MAGE-A4 CD8 TCR T cells. Additionally, circulating cytokines were measured and evaluated in association with cytokine release syndrome (CRS) and other adverse events (AEs). Additionally, transdextrin persistence was assessed by determining the persistence of serum levels of MAGE-A4 CD8 TCR-modified T cells, measured by the MAGE-A4 TCR vector copy number and the number of MAGE-A4 CD8 TCR transdextrinsed T cells after MAGE-A4 CD8 TCR T cell infusion. The mean expression of specific surface markers on genetically modified T cells in the target blood and tumors was measured by fluorescence intensity. The killing and cytokine profiles of genetically modified T cells were evaluated using flow cytometry in the blood and tumors. The biomarkers for the target samples include polymorphisms in cytokine genes and cytokine production.

[0145] To evaluate the antitumor activity of MAGE-A4 CD8 TCR T cells, the following endpoint was monitored according to RECIST v1.1: overall response rate (ORR), defined as the proportion of subjects with confirmed complete response (CR) or partial response (PR). Additional endpoints monitored included duration of response (DoR), stable disease (SD), progression-free survival (PFS), and overall survival (OS). The efficacy of the treatment was evaluated by assessing the duration of response and overall survival. (a) The date of the first T-cell infusion administration, and the date of evaluation of the efficacy of the treatment by assessment of the first documented evidence for CR or PR, and the time to the first response. (b) The date of the first documented evidence of complete response (CR) or partial response (PR) to the first documented disease progression or death due to any cause. (c) The date of the first documented evidence of stable disease (SD) until the first documented disease progression or death due to any cause. (d) The date of the first T-cell infusion, and the earliest date of death due to the disease, progression, or any other cause. (e) Between the day of the first T cell infusion and the day of death due to any cause We also assessed the intervals related to this.

[0146] Evaluation of the efficacy of treatment by the number and percentage of subjects with any long-term follow-up adverse events (AEs), malignancies, neurological disorders, rheumatic or other autoimmune disorders, hematological disorders, or infections.

[0147] Safety and tolerability responses were additionally monitored through laboratory assessments including chemistry, hematology, and coagulation, as well as adverse events (AEs) including anti-MAGE-A4 TCR antibodies, severe adverse events (SAEs), dose-limiting toxicity (DLTs), NCI CTCAEs, and optimally tolerable dose ranges, and PCR-based assays for the persistence of genetically modified T cells in the periphery and the retention of heterologous TCR expression in T cell PBMCs.

[0148] result The data in Figure 1 are CT scans of a 31-year-old male with HER2-negative stage 4 adenocarcinoma of GEJ, who had received prior unsuccessful chemotherapy, targeted therapy, and immunotherapy regimens (ramucirumab + paclitaxel, atezolizumab + BL-8040, FLOT / FOLFIRI chemotherapy). The subject had moderate MAGE-A4 expression (IHS 3+) and a large disease burden at baseline; baseline lesion SLD was 66 cm, and the subject was offered an initial infusion of approximately 10 billion MAGE-A4 CD8 TCR T cells, with minimal adverse reactions consistent with those typically experienced by cancer patients receiving cytotoxic chemotherapy and / or cancer immunotherapy. The tumor showed a reduction of over 42% at week 8, as measured by RECIST 1.1 and by the sum of the diameter of the targeted lesion over a measurement period of several weeks following the day of T cell infusion. This progressed to a reduction of over 51% at week 18, as shown in Figure 2. The data support the efficacy of the 18-week response in terms of tumor size reduction (51% reduction) for esophagogastric junction (EGJ) cancer treatment. Data were presented for aortic and vena cava lymph nodes, and equivalent data were obtained by CT for periportal lymph nodes, ascites, and nonmeaurable peritoneal metastases (met), with equivalent reductions from baseline observed for aortic and vena cava lymph nodes.

[0149] Over a period of 8–12 weeks post-infusion, the absolute concentration of transduced lymphocytes (lymphocytocytes), including MAGE-A4 CD8 TCR T cells, was maintained at high levels (46–53%), demonstrating the persistence of therapeutic T cells.

[0150] array Sequence ID 1, MAGE A4

[0151] [ka]

[0152] Sequence ID No. 2, MAGE A4 peptide GVYDGREHTV

[0153] Sequence ID 3; (CD8α) CDR in bold and underlined, signal sequence in italics and underlined

[0154] [ka]

[0155] Sequence ID 4; (CD8α)

[0156] [ka]

[0157] Sequence ID 5; (MAGE A4 TCR α chain) CDR in bold and underlined

[0158] [ka]

[0159] Sequence ID 6; (MAGE A4 TCR α-chain coding sequence)

[0160] [ka]

[0161] Sequence ID 7; (MAGE A4 TCR β chain) CDR in bold and underlined

[0162] [ka]

[0163] Sequence ID 8; (MAGE A4 TCR β-chain coding sequence)

[0164] [ka]

[0165] Sequence ID 9; (MAGE A4 TCR α-chain variable region) 136AA - CDR (Bold and underlined) MKKHLTTFLVILWLYFYRGNGKNQVEQSPQSLIILEGKNCTLQCNYTVSPFSNLRWYKQDTGRGPVSLTILTFSENTKSNGRYTATLDADTKQSSLHITASQLSDSASYICVVSGGTDSWGKLQFGAGTQVVVTPD

[0166] Sequence ID 10; (MAGE A4 TCR β-chain variable region) 133AA - CDR (Bold and underlined) MASLLFFCGAFYLLGTGSMDADVTQTPRNRITKTGKRIMLECSQTKGHDRMYWYRQDPGLGLRLIYYSFDVKDINKGEISDGYSVSRQAQAKFSLSLESAIPNQTALYFCATSGQGAYEEQFFGPGTRLTVLE

[0167] Sequence ID 11; CDR1 MAGE A4 TCR α chain, (residues 48-53) VSFSSN

[0168] Sequence ID 12; CDR2 MAGE A4 TCR α chain, (residues 71-76) LTFSEN

[0169] Sequence ID 13; CDR3 MAGE A4 TCR α chain, (residues 111-125) CVVSGGTDSWGKLQF

[0170] Sequence ID 14; CDR1 MAGE A4 TCR β chain, (residues 46-50) KGHDR

[0171] Sequence ID 15; CDR2 MAGE A4 TCR β chain, (residues 68-73) SFDVKD

[0172] Sequence ID 16; CDR3 MAGE A4 TCR β chain, (residues 110-123) CATSGQGAYEEQFF

[0173] Sequence ID 17; CDR1 CD8α (residues 45-53) VLLSNPTSG

[0174] Sequence ID 18; CDR2 CD8α (residues 72-79) YLSQNKPK

[0175] Sequence ID 19; CDR3 CD8α (residues 118-123) LSNSIM

Claims

1. 1. An engineered immunoresponsive cell that expresses or presents a heterologous T cell receptor (TCR) that binds a peptide antigen of GVYDGREHTV, MAGE A4 comprising SEQ ID NO:2, for use in a method of treating, preventing or delaying the progression of cancer and / or tumor in a subject, said method comprising administering to said subject a therapeutic regimen comprising an effective amount of said engineered immunoresponsive cell, wherein said cancer and / or tumor is a gastroesophageal cancer and / or tumor. (a) the heterologous TCR specifically and / or selectively binds to the peptide antigen; (b) the peptide antigen is associated with and / or presented by a gastroesophageal cancer and / or tumor and / or cancer cells or tissues, and / or (c) the cancer and / or tumor is a MAGE A4-expressing cancer and / or tumor and / or expresses MAGE A4 or a peptide antigen thereof, or a peptide antigen of MAGE A4, including GVYDGREHTV, SEQ ID NO:2; The modified immunoresponsive cell of claim 1. (a) the heterologous TCR specifically and / or selectively binds to the peptide antigen; (b) the peptide antigen is a peptide presenting molecule, optionally wherein the peptide presenting molecule is (i) complexed with a major histocompatibility complex (MHC) or a human leukocyte antigen (HLA), optionally class I or class II, or (ii) optionally HLA*02, HLA-A*02:01, HLA-A*02:02, HLA-A*02:03, HLA-A*02:04, HLA- HLA-A*02:05, HLA-A*02:06, HLA-A*02:642, or HLA-A*02:07, preferably HLA-A*02:01 or HLA-A*02, and optionally said xenogenous TCR specifically and / or selectively binds to said peptide antigen and / or said peptide presenting molecule, and / or complexes thereof, or (c) the peptide antigen is presented independent of a peptide-presenting molecule; The modified immunoresponsive cell of claim 1 or 2.

4. The heterologous TCR, (a) a TCR alpha chain variable domain and a TCR beta chain variable domain; (i) the alpha chain variable domain has the sequence VSPFSN (αCDR1), SEQ ID NO:11, or amino acid residues 48-53 of SEQ ID NO:5, or a sequence having at least 50% sequence identity thereto; LTFSEN (αCDR2), SEQ ID NO: 12, or amino acid residues 71-76 of SEQ ID NO: 5, or a sequence having at least 50% sequence identity thereto; and CVVSGGTDSWGKLQF (αCDR3), SEQ ID NO:13, or amino acid residues 111-125 of SEQ ID NO:5, or a sequence having at least 50% sequence identity thereto. and (ii) the beta chain variable domain has the sequence KGHDR (βCDR1), SEQ ID NO: 14, or amino acid residues 46-50 of SEQ ID NO: 7, or a sequence having at least 50% sequence identity thereto; SFDVKD (βCDR2), SEQ ID NO: 15, or amino acid residues 68-73 of SEQ ID NO: 7, or a sequence having at least 50% sequence identity thereto; and CATSGQGAYEEQFF (βCDR3), SEQ ID NO: 16, or amino acid residues 110-123 of SEQ ID NO: 7, or a sequence having at least 50% sequence identity thereto. and / or (b) the heterologous TCR is (i) the alpha chain variable domain comprises an amino acid sequence having at least 80% identity to SEQ ID NO: 9, and / or the beta chain variable domain comprises an amino acid sequence having at least 80% identity to SEQ ID NO: 10; (ii) the alpha chain variable domain comprises an amino acid sequence comprising SEQ ID NO:9, and / or the beta chain variable domain comprises SEQ ID NO:10; (iii) the alpha chain comprises an amino acid sequence having at least 80% identity to SEQ ID NO:5, and / or the beta chain comprises an amino acid sequence having at least 80% identity to SEQ ID NO:6, or (iv) the alpha chain comprises an amino acid sequence comprising SEQ ID NO:5, and / or the beta chain comprises an amino acid sequence comprising SEQ ID NO:6; The modified immunoresponsive cell of claim 1 .

5. The modified immunoresponsive cell expressing or presenting a heterologous TCR further expresses or presents a heterologous co-receptor, and optionally (a) the heterologous co-receptor is a CD8 co-receptor; (b) the heterologous co-receptor is a CD8 co-receptor, and the CD8 co-receptor is a heterodimer or a homodimer, a CD8αb heterodimer or a CD8αα homodimer; (c) the heterologous co-receptor is a heterologous CD8 co-receptor, the heterologous CD8 co-receptor comprising: (i) an amino acid sequence VLLSNPTSG, a CDR1 having at least 80% sequence identity with SEQ ID NO:17, an amino acid sequence YLSQNKPK, a CDR2 having at least 80% sequence identity with SEQ ID NO:18, and an amino acid sequence LSNSIM, a CDR3 having at least 80% sequence identity with SEQ ID NO:19; (ii) an amino acid sequence VLLSNPTSG, a CDR1 of SEQ ID NO: 17, an amino acid sequence YLSQNKPK, a CDR2 of SEQ ID NO: 18, and an amino acid sequence LSNSIM, a CDR3 of SEQ ID NO: 19; (iii) an amino acid sequence having at least 80% sequence identity to amino acid numbers 22 to 235 of SEQ ID NO: 3, or 22 to 135 of SEQ ID NO: 3; or (iv) an amino acid sequence having 100% sequence identity with amino acid numbers 22 to 235 of SEQ ID NO: 3, or 22 to 135 of SEQ ID NO: 3 Any one of the following: The modified immunoresponsive cell of claim 1 . (a) the modified immunoresponsive cell further expresses or presents a heterologous costimulatory ligand, optionally 4-1BBL or CD80; (b) the modified immunoresponsive cell is (i) a B cell, a T cell, or a natural killer (NK) cell; (ii) a T cell, optionally a CD4 + T cell and / or a CD8 + T cell; and / or (c) the modified immunoresponsive cells are a population of CD4+ T cells; or CD8+ T cells, or a mixed population of CD4+ and CD8+ T cells. The modified immunoresponsive cell of claim 1 .

7. The modified immunoresponsive cell, (a) administered continuously or intermittently; (b) administered as multiple doses or as a single dose; and / or (c) administered intravenously or by intravenous infusion; The modified immunoresponsive cell of any one of claims 1 to 6.

8. The modified immunoresponsive cell, (a) administered in multiple doses or a single dose, said single dose or multiple doses being administered in one or more dosing cycles, optionally said doses may be fixed or variable doses; (b) administered in a dose of about 500 million to about 1 billion cells, about 2 billion to about 5 billion cells, or about 6 billion to about 10 billion cells; and / or (c)(i) a single dose in each of one or more dosing cycles; (ii) one or more doses in each of one or more dosing cycles; (iii) a single dose on day 1 of each of one or more dosing cycles; (iv) one or more doses in each of one or more dosing cycles, with at least one dose on day 1 of each cycle; (v) one or more doses in each of one or more dosing cycles, with at least one dose on day 1 of each cycle; or (vi) Single Dose Administered as Optionally, the dosing cycle is for 2-6 months or during disease progression. The modified immunoresponsive cell of any one of claims 1 to 7.

9. (i) The dosing cycle comprises: (a) disease progression following a prior administration of modified immunoresponsive cells, and 12 weeks or more after a prior administration of modified immunoresponsive cells; In (b) the tumor and / or cancer expresses MAGE-A4 and / or a peptide antigen thereof, and / or (c) MAGE-A4 and / or its peptide antigens are detected in the subject's biological sample and / or are above the normal range, or (ii) the dosing cycle comprises: (a) a complete or partial response following the previous administration of modified immunoresponsive cells, or (b) stable disease for a period of more than or equal to 4 months following the previous administration of modified immunoresponsive cells, followed by disease progression; and (c) more than or equal to 12 weeks following the previous administration of modified immunoresponsive cells. In (d) the tumor and / or cancer expresses MAGE-A4 and / or a peptide antigen thereof, and / or (e) MAGE-A4 and / or its peptide antigens are detected in the subject's biological sample and / or are above the normal range; The modified immunoresponsive cell of claim 8.

10. (a) prior to treatment, the subject has measurable disease according to Eastern Cooperative Oncology Group (ECOG) 0-1 and / or Response Evaluation Criteria in Solid Tumors (RECIST) 1.1 and / or histologically confirmed gastroesophageal cancer and / or tumors; (b) prior to treatment, the subject: (i) the HLA-A genotype is HLA-A*02:07b as the only HLA-A*02 allele; (ii) the HLA-A genotype is HLA-A* with any A*02 null allele as the only HLA-A*02 allele; or (iii) Symptomatic CNS metastases and / or, (c) the subject is intolerant to standard of care treatment, optionally systemic platinum-based chemotherapy treatment; The modified immunoresponsive cell of any one of claims 1 to 9.

11. (a) the gastroesophageal cancer and / or tumor has previously failed treatment with standard of care therapy, optionally with systemic platinum-based chemotherapy; (b) the gastroesophageal cancer and / or tumor has previously failed treatment with any of surgery (resection), radiation therapy, targeted therapy, immunotherapy, or chemotherapy, or combination chemotherapy with surgery (resection), radiation therapy, radiation therapy, targeted therapy, checkpoint inhibitor, or immunotherapy; (c) the subject has a primary cancer, a secondary cancer, a relapsing or refractory cancer, a recurrent or locally recurrent cancer, a metastatic cancer, an unresectable or locally localized cancer with no surgical or radiotherapy options, or an inoperable cancer, or the gastroesophageal cancer and / or tumor is a primary cancer, a secondary cancer, a relapsing or refractory cancer, a recurrent or locally recurrent cancer, a metastatic cancer, an unresectable or locally localized cancer with no surgical or radiotherapy options, or an inoperable cancer, optionally wherein the cancer is not suitable for transplantation or locoregional therapy, and / or (d) the gastroesophageal cancer and / or tumor is any of esophageal squamous cell carcinoma (ESCC), esophageal adenocarcinoma (EAC), esophagogastric junction cancer, carcinoma, adenocarcinoma, or tumor (EGJ), or stomach or gastric cancer, carcinoma, or tumor, optionally metastatic and / or progressive and / or locally progressive and / or recurrent ESCC, EAC, EGJ, or stomach or gastric cancer, carcinoma, or tumor; The modified immunoresponsive cell of any one of claims 1 to 10.

12. The method of claim 1, wherein the subject receives lymphodepleting chemotherapy prior to administration of the modified immunoresponsive cells. Optionally, said lymphodepleting chemotherapy comprises: (i) comprising administration of cyclophosphamide and fludarabine, optionally at a dose of 500 mg / m2 / d x 3d cyclophosphamide and 20 mg / m2 / d x 3d fludarabine, or at a dose of 600 mg / m2 / d x 3d cyclophosphamide and 30 mg / m2 / d x 4d fludarabine; and / or (ii) is administered 7-5 or 7-4 days prior to administration of the engineered immunoresponsive cells expressing or displaying a heterologous T cell receptor (TCR); The modified immunoresponsive cell of any one of claims 1 to 11.

13. The object, (a) have not received prior treatment for cancer and / or tumors; or (b) has undergone a prior cancer and / or tumor treatment and / or has not responded to a prior cancer and / or tumor treatment, optionally wherein said prior treatment is for gastroesophageal cancer and / or tumor; Optionally, said prior treatment comprises systemic and / or local therapy, optionally any one or more of surgery, radiation therapy, cryotherapy, laser therapy, local therapy, and / or any one or more of systemic therapy, such as chemotherapy, hormone therapy, targeted drugs, targeted chemotherapy, or immunotherapy; The modified immunoresponsive cell of any one of claims 1 to 12.

14. The subject has undergone a prior cancer and / or tumor treatment and / or has not responded to a prior cancer and / or tumor treatment, optionally the prior treatment being for a gastroesophageal cancer and / or tumor, and the prior treatment being: (i) a PD-L1 binding antagonist or a PD-1 binding antagonist, optionally wherein the PD-1 axis binding antagonist or the PD-L1 binding antagonist is an antibody; (ii) an epidermal growth factor receptor antagonist, optionally comprising any of cetuximab, erlotinib, gefitinib, or afatinib, or a vascular endothelial growth factor (VEGF) inhibitor, such as, for example, ramucirumab or bevacizumab, or an EGFR inhibitor antibody, such as panitumumab or cetuximab, or a human hepatocyte growth factor HGF and / or Met inhibitor, such as onartuzumab or rilotumumab; (iii) chemotherapy optionally comprising a platinum compound selected from any of lipoplatin, cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, satraplatin, picoplatin; (iv) chemotherapy comprising a chemotherapeutic agent selected from any of methotrexate, capecitabine, taxanes, anthracyclines, paclitaxel, docetaxel, paclitaxel protein-bound particles, doxorubicin, epirubicin, 5-fluorouracil, cyclophosphamide, afatinib, vincristine, etoposide, or a combination thereof; or (v) chemotherapy comprising a chemotherapeutic agent selected from the following: FEC: 5-fluorouracil, epirubicin, cyclophosphamide; FAC: 5-fluorouracil, doxorubicin, cyclophosphamide; AC: doxorubicin, cyclophosphamide; EC: epirubicin, cyclophosphamide; Optionally, the subject (a) has not received prior treatment in a recurrence less than or equal to 12 months since the last treatment, or less than or equal to 6 months since the last treatment, or (b) has not received any prior adjuvant therapy (e.g., surgery followed by radiation and / or chemotherapy) or locoregional therapy in a recurrence less than or equal to 12 months since the last treatment, or less than or equal to 6 months since the last treatment. The modified immunoresponsive cell of claim 13.

15. The treatment, as compared to a placebo treatment, or to prior treatment, or to no treatment, or to a treatment including standard of care, optionally systemic platinum-based chemotherapy treatment, (a) progression-free survival, (b) time to progression, (c) duration of response, (d) overall survival; (e) objective response or objective response rate, (f) overall response or overall response rate, (g) partial response or partial response rate, (h) complete response or complete response rate; (i) Rate of stable disease or median stable disease (j) median progression-free survival, (k) median time to progression; (l) median duration of response; (m) median overall survival; (n) median objective response or median objective response rate; (o) median overall response or median overall response rate; (p) median partial response or median partial response rate; (q) the median complete response or the median complete response, or (r) Median stable disease rate or median stable disease rate 15. The modified immunoresponsive cell of claim 1, which effectively extends or improves

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