Modified T cells for use in the treatment of gastroesophageal cancer

Abstract

The present disclosure relates to methods of treating gastroesophageal cancer and populations of modified T cells expressing heterologous TCRs for use in such methods.

Description

[Technical Field]

[0001] The present disclosure relates to methods of treating gastroesophageal cancer and populations of modified T cells expressing heterologous TCRs for use in such methods. [Background technology]

[0002] Gastroesophageal cancer includes cancer of the esophagus, gastroesophageal junction, or stomach. Gastroesophageal cancer is sometimes known as gastroesophageal tumors.

[0003] Esophageal Cancer: Esophageal cancer is the sixth leading cause of cancer-related deaths worldwide. Esophageal cancer generally arises from the esophageal epithelium and is classified into one of 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 cancer, carcinoma, or adenocarcinoma of the gastroesophageal junction (EGJ), a cancer of the lower esophagus that is often associated with Barrett's esophagus. EGJ is a highly mutated and heterogeneous disease with an increasing number of somatic mutations in many genes, including CR2, HGF, FGFR4, ESRRB, TP53, SYNE1, and ARID1A. Treatment options for gastroesophageal junction adenocarcinoma are limited, and the overall prognosis is extremely poor. Esophageal squamous cell carcinoma (ESCC) accounts for 60–70% of all esophageal cancer cases worldwide, with an additional 20–30% of cases being esophageal adenocarcinoma (EAC). Other less common forms of cancer include neuroendocrine carcinoma, melanoma, leiomyosarcoma, carcinoid, and lymphoma. Esophageal cancer generally has a poor prognosis, with an overall 5-year survival rate of approximately 15% in the United States, with most patients dying within one year of diagnosis. Recent data from England and Wales indicate that approximately 10% of patients with esophageal cancer survive at least 10 years. While the disease's prevalence is explained by its poor prognosis, the low survival rate is also related to low rates of early detection, as most patients have advanced disease by the time the first symptoms, such as difficulty swallowing, appear. In the United States, esophageal cancer is the seventh leading cause of cancer death among men. Definitive treatment options for localized disease involve a combination of surgery, radiation, and chemotherapy. Metastatic or recurrent disease is typically managed palliatively with stent placement to relieve symptoms and facilitate swallowing, radiation, and chemotherapy.

[0004] Gastric Cancer: Gastric cancer is sometimes known as cancer of the stomach. Gastric / stomach cancer is closely associated with tobacco, alcohol, and Helicobacter pylori, as well as eating salted or pickled foods. Most gastric cancers are adenocarcinomas. Other types of gastric cancer include squamous cell carcinoma, gastrointestinal stromal tumor (GIST), non-Hodgkin's lymphoma, and neuroendocrine tumor (NET). Recent data from England indicate that approximately 15% of people with gastric cancer survive at least 10 years. Globally, gastric cancer is the third leading cause of cancer death, occurring twice as often in men as in women. In the United States, it accounts for 9% of deaths, with a 5-year survival rate of 31.5%.

[0005] Gastroesophageal cancer is staged according to the TNM classification system, where T is the size and shape of the tumor, N is the presence or absence of lymph node metastasis, and M is the presence or absence of distant metastasis. The T, N, and M features are combined to determine the cancer's "stage," ranging from I to IVB.

[0006] In general, gastroesophageal adenocarcinoma and squamous cell carcinoma are treated in the same way. Treatment typically includes a surgical component when possible. Surgery may be combined with chemotherapy (using drugs such as 5-fluorouracil, cisplatin, epirubicin, etoposide, docetaxel, oxaliplatin, capecitabine, or irinotecan), radiation therapy, or chemoradiotherapy, which may be administered before or after surgery. For example, targeted and / or immunotherapy using the human epidermal growth factor receptor 2 (HER2) inhibitor trastuzumab can be combined with chemotherapy for HER2-overexpressing cancers, such as adenocarcinoma.

[0007] Surgical resection and radiation therapy (including 3D conformal radiation therapy, intensity-modulated radiation therapy, particle therapy, and brachytherapy) or combination chemotherapy regimens are the mainstay of treatment for most gastroesophageal cancers and are the standard of care for tumors with local metastases (stage III or IV). For early-stage primary cancers without local metastases (stage I or II), surgery alone may be sufficient. Typical chemotherapy agents include paclitaxel in combination with carboplatin and fluorouracil in combination with oxaliplatin / cisplatin. Oxaliplatin is generally preferred over cisplatin in first-line treatment due to its lower toxicity. Docetaxel, capecitabine, and irinotecan are also frequently used in neoadjuvant / postoperative chemotherapy or chemoradiation regimens.

[0008] Immune checkpoint blockade offers additional treatment options. Trastuzumab is recommended in combination with oxaliplatin / cisplatin and a fluoropyrimidine (such as fluorouracil or capecitabine) as first-line treatment for unresectable locally advanced, relapsed, or metastatic disease. Pembrolizumab and nivolumab are preferred in combination with oxaliplatin / cisplatin and a fluoropyrimidine for the treatment of HER2-negative cancers.

[0009] New therapies for treating, preventing, and / or slowing the progression of gastroesophageal cancer are desirable. Summary of the Invention

[0010] The present inventors have demonstrated that the use of engineered T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor capable of binding to a peptide antigen of MAGE-A4 is advantageous as a treatment for gastroesophageal cancer. Engineered T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor capable of binding to a peptide antigen of MAGE-A4 can be included, for example, in early (e.g., first-line, second-line, or even third-line) treatment for gastroesophageal cancer. The present inventors have devised an exemplary treatment regimen in light of these considerations.

[0011] Including T cell therapy in advanced treatment options may alter the tumor microenvironment by infiltrating the tumor and potentially enhance and sustain T cell activation by leveraging a broader immune response. This may improve the frequency, depth, and durability of responses. Furthermore, T cell therapy in advanced treatment options has the advantage of reaching healthier patients, who have a more favorable tumor microenvironment and are more likely to harvest T cells. Furthermore, healthier patients may be more likely to respond to treatment and experience fewer undesirable effects, such as cytokine release syndrome (CRS) and cytopenias.

[0012] T cell therapy can be included in a combination therapy, such as a combination therapy that includes an additional anti-cancer therapy (such as chemotherapy) and / or a checkpoint inhibitor. Including the engineered T cells in the combination therapy can be advantageous because it may allow for a reduction in the dose of the additional anti-cancer therapy or checkpoint inhibitor. This may result in a reduction in CRS and / or cytopenias, particularly when the anti-cancer therapy is chemotherapy.

[0013] Accordingly, the present disclosure provides a method of treating gastroesophageal cancer in an individual, the method comprising administering to the individual a population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor (TCR) capable of binding to a peptide antigen of MAGE-A4.

[0014] The present disclosure also provides a population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor capable of binding to a peptide antigen of MAGE-A4 for use in the methods of the disclosure. [Brief explanation of the drawings]

[0015] [Figure 1] Exemplary treatment regimens for gastroesophageal cancer that has recurred after first-line treatment. [Figure 2] Exemplary treatment regimens for gastroesophageal cancer that has recurred after second-line treatment. [Figure 3]Alternative expressions of exemplary treatment regimens for (1) gastroesophageal cancer that has recurred after first-line treatment and (2) gastroesophageal cancer that has recurred after second-line treatment. [Figure 4] Exemplary treatment regimens for gastroesophageal cancer that has (A) recurred after treatment with curative intent for locally advanced disease or (B) is first diagnosed as unresectable locally advanced or metastatic disease. [Figure 5] Alternative representations of exemplary treatment regimens for gastroesophageal cancer that has (A) recurred after treatment with curative intent for locally advanced disease or (B) is first diagnosed as unresectable locally advanced or metastatic disease. [Figure 6] Efficacy of ADP-A2M4CD8 in patients with advanced esophageal, gastroesophageal junction, or gastric cancer. Figure 6A: Change in baseline sum of longest diameters (SLD) of target lesions in individual patients. Figure 6B: Change in baseline target SLD by week after T cell infusion. One unevaluable patient is not shown. Data represent change from baseline in SLD until progression or prior to surgical resection. Investigator-assessed best overall response rate is shown according to RECIST v1.1. Combo refers to patients in the nivolumab combination group. EGJ: gastroesophageal junction. ESO: esophagus. G: stomach. PD: progressive disease. PR: partial response. RECIST: Response Evaluation Criteria in Solid Tumors. SD: stable disease. DETAILED DESCRIPTION OF THE INVENTION

[0016] It is to be understood that the various applications of the disclosed methods and products can be tailored to the particular needs of the art, and that the terminology used herein is for the purpose of describing particular embodiments of the disclosure only, and is not intended to be limiting.

[0017] All publications, patents, and patent applications cited herein, whether supra or infra, are hereby incorporated by reference in their entirety.

[0018] general definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0019] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a TCR" includes "TCRs," reference to "an antibody" includes two or more such antibodies, etc.

[0020] In general, the term "comprising" is intended to mean including, but not limited to. For example, the phrase "a method comprising administering a population of modified T cells" should be interpreted to mean that the method includes administering such a population, but may also include additional steps, such as administering an additional therapeutic agent.

[0021] In some aspects of the disclosure, the word "comprising" is replaced with the phrase "consisting of." The term "consisting of" is intended to be restrictive. For example, the phrase "a method consisting of administering a population of modified T cells" should be interpreted to mean that the method includes the step of administering such population, and that there are no additional steps.

[0022] The terms "protein" and "polypeptide" are used interchangeably herein and are intended to refer to polymeric chains of amino acids of any length.

[0023] Typically, the term "about" is used to refer to a value that is within ±10% (such as within ±5% or ±2%) of the value that follows it.

[0024] As used herein, the terms "relapsed" and "recurrent" cancer are interchangeable.

[0025] For purposes of this disclosure, to determine the percent identity of two sequences (such as two polynucleotide or two polypeptide sequences), the sequences are aligned for optimal comparison purposes (e.g., gaps may be introduced into the first sequence to achieve optimal alignment with the second sequence). Nucleotide residues are then compared at nucleotide position. If a position in the first sequence is occupied by the same nucleotide residue as the corresponding position in the second sequence, the nucleotides are identical at that position. The percent identity between two sequences is a function of the number of identical positions shared by the sequences (i.e., percent identity = number of identical positions / total number of positions in the reference sequence × 100).

[0026] Typically, sequence comparison is performed over the entire length of a reference sequence. For example, if a user wants to determine whether a given ("test") sequence has a certain percent identity to SEQ ID NO:X, SEQ ID NO:X would be the reference sequence. For example, to assess whether a sequence is at least 80% identical to SEQ ID NO:X (an example of a reference sequence), one skilled in the art would align the entire length of SEQ ID NO:X and determine how many positions in the test sequence are identical to SEQ ID NO:X. If at least 80% of the positions are identical, the test sequence is at least 80% identical to SEQ ID NO:X. If a sequence is shorter than SEQ ID NO:X, gaps or missing positions should be considered non-identical positions.

[0027] Those skilled in the art are aware of various computer programs available to determine homology or identity between two sequences. For example, the comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm.

[0028] Cancer treatment methods The present disclosure provides a method of treating gastroesophageal cancer in an individual, the method comprising administering to the individual a population of engineered T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor (TCR) capable of binding to a peptide antigen of MAGE-A4. As described above, the inventors have demonstrated that such a method is advantageous. Engineered T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor capable of binding to a peptide antigen of MAGE-A4 may be included, for example, in second- or third-line treatments for gastroesophageal cancer.

[0029] In the context of the present disclosure, treating gastroesophageal cancer may also include preventing and / or slowing the progression of gastroesophageal cancer.

[0030] Gastroesophageal cancer in individuals The disclosed methods are for treating gastroesophageal cancer in an individual. The individual is preferably a human. The individual may also be a non-human mammal, such as a mouse, rat, rabbit, cat, dog, pig, cow, or horse.

[0031] Gastroesophageal cancer may express MAGE-A4. Expression of MAGE-A4 has been reported in gastroesophageal cancer. For example, approximately 20% of solid tumors in gastroesophageal cancer express MAGE-A4, and 40-45% of these express HLA-A. * 02 also appears.

[0032] At least 1%, e.g., at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, or at least 80% of gastroesophageal cancer cells from an individual may express MAGE-A4. The percentage of cells expressing MAGE-A4 can be determined by any means known to those skilled in the art, such as immunohistochemistry (IHC), flow cytometry, or enzyme-linked immunosorbent assay (ELISA).

[0033] MAGE-A4 expression in gastroesophageal cancer can have an intensity of 1+ or greater (≧), for example, ≧2+ or ≧3+. Intensity scores can be assessed by IHC staining of tumors and are scored as follows: negative = no staining or staining in ≦10% or less of the stained cells; 1+ = incomplete staining in ≧10% of the stained cells; 2+ = weak to moderate staining in ≧10% of the stained cells; strong and complete staining in ≧10% of the stained cells.

[0034] The gastroesophageal cancer may be a gastroesophageal tumor. The gastroesophageal cancer may be, for example, a solid tumor. The gastroesophageal cancer may be, for example, a squamous cell carcinoma, an adenocarcinoma, a gastrointestinal stromal tumor (GIST), a non-Hodgkin's lymphoma, a neuroendocrine tumor (NET), a melanoma, a leiomyosarcoma, a carcinoid tumor, or a lymphoma. The gastroesophageal cancer may be, for example, a squamous cell carcinoma or an adenocarcinoma.

[0035] The gastroesophageal cancer may be, for example, esophageal cancer. The esophageal cancer may be, for example, esophageal squamous cell carcinoma (ESCC). The esophageal cancer may be, for example, esophageal adenocarcinoma (EAC). The gastroesophageal cancer may be, for example, gastroesophageal junction cancer (EGJ; also known as gastroesophageal junction cancer or GOJ). The gastroesophageal cancer may be, for example, cervical esophageal cancer. The gastroesophageal cancer may be, for example, gastric cancer. The gastric cancer may be, for example, gastric adenocarcinoma. The gastroesophageal cancer may be associated with gastroesophageal reflux disease (GERD) or Barrett's esophagus.

[0036] The gastroesophageal cancer may be, for example, primary or secondary gastroesophageal cancer. The gastroesophageal cancer may be, for example, recurrent or recurrent, unresectable, locally advanced, and / or metastatic gastroesophageal cancer. For example, the gastroesophageal cancer may be unsuitable for treatment by surgical resection or radiation therapy.

[0037] The gastroesophageal cancer may be recurrent gastroesophageal cancer. The recurrent gastroesophageal cancer may be, for example, locally advanced recurrence or metastatic recurrence. The gastroesophageal cancer may be, for example, a recurrence after treatment with curative intent. Thus, the individual may be a cancer patient who has undergone treatment with curative intent for gastroesophageal cancer. Such cancer patients form a subset of gastroesophageal cancer patients that is well recognized in the art.

[0038] In the context of this disclosure, "selection" of a treatment may refer to a treatment regimen for a cancer that has failed curative treatment or for which curative treatment may be inappropriate. Curative treatment may refer to any treatment that has the potential to cure an individual's cancer. Curative treatment may refer, for example, to a therapy administered to an individual to attempt to cure the cancer. Curative treatment may occur before "selection" of a treatment for a relapsed or spread cancer.

[0039] Failure of a curative treatment can refer, for example, to the failure of a curative treatment to eliminate or induce remission of the cancer. For example, failure of a curative treatment can result in recurrence or spread of the cancer, such as local invasion or metastasis. When the cancer has progressed, for example, when the cancer is locally advanced or metastatic, curative treatment may be inappropriate. Thus, a "selection" of treatment can refer to a treatment regimen for locally advanced cancer, metastatic cancer, or recurrent cancer. A "selection" of treatment can refer to a treatment regimen for recurrent or spread cancer.

[0040] A "choice" of treatment may be, for example, a first-line treatment for cancer. In other words, the treatment regimen may be an initial treatment regimen adopted for cancer after failure or after treatment with curative intent. For example, the treatment regimen may be an initial treatment regimen adopted for cancer after relapse or spread of cancer. For example, the treatment regimen may be an initial treatment regimen adopted for cancer after local progression / invasion, metastasis, or recurrence. A "choice" of treatment may be, for example, a second-line treatment for cancer. In other words, the treatment regimen may be, for example, a second treatment regimen adopted for cancer after failure of the first treatment regimen. Failure of the first treatment regimen may result in, for example, relapse or further spread of cancer. Failure of the first treatment regimen may result in, for example, local progression / invasion, metastasis, or recurrence. A choice of treatment may be, for example, a third-line treatment for cancer. In other words, the treatment regimen may be, for example, a third treatment regimen adopted for cancer after failure of the second treatment regimen. Failure of a second treatment regimen may result, for example, in recurrence or further spread of the cancer.Failure of a second treatment regimen may result, for example, in local progression / invasion, metastasis, or recurrence.

[0041] The gastroesophageal cancer may be, for example, a recurrence after first-line treatment of gastroesophageal cancer. Recurrence may refer to, for example, recurrence, local invasion, and / or metastasis. First-line treatments are described in more detail below. However, by way of example, the first-line treatment may be a treatment that has already been described as an approved or "standard of care" first-line treatment for one or more gastroesophageal cancers. In this context, approval may relate to approval by, for example, the FDA, EMA, or MHRA. "Standard of care" first-line treatments for gastroesophageal cancer are well known in the art and are described in publicly available clinical guidelines, such as those provided by the National Comprehensive Cancer Network.

[0042] First-line "standard of care" treatments for operable esophageal cancer (or for esophageal cancers amenable to local therapy) can include, for example, (i) surgical resection, (ii) radiation therapy, and / or (iii) systemic therapy. For example, first-line "standard of care" treatments for operable esophageal cancer can include (i); (ii); (iii); (i) and (ii); (i) and (iii); (ii) and (iii); or (i), (ii), and (iii). Operable esophageal cancer can include, for example, pTis tumors (primary tumors confined intraepithelially by a basement membrane) and pT1 tumors (primary tumors invading the lamina propria, muscularis mucosa, or submucosa). Surgical treatments for such cancers can include, for example, endoscopic resection, endoscopic resection followed by ablation, or esophagectomy.

[0043] For inoperable gastroesophageal cancer (or gastroesophageal cancer for which a surgical approach is not feasible, such as gastroesophageal junction adenocarcinoma and cervical esophageal cancer), the "standard of care" first-line treatment may include, for example, definitive chemoradiotherapy. Alternatively, other anti-cancer therapies (such as systemic therapies) may be used.

[0044] The gastroesophageal cancer treated by the methods of the present disclosure may have recurred after a second-line or subsequent-line treatment for gastroesophageal cancer. In other words, the gastroesophageal cancer may have recurred for the first time after a first-line treatment and then recurred a second time after a second-line treatment. The gastroesophageal cancer may also have progressed to a subsequent-line treatment. Recurrence may refer to, for example, relapse, local invasion, and / or metastasis. First-line and second-line treatments are described in more detail below. However, by way of example, the first-line treatment may be one or more approved or "standard of care" treatments for gastroesophageal cancer, as described above. Also, by way of example, the second-line treatment may be one or more approved or "standard of care" treatments for gastroesophageal cancer, as described above. In this context, approval may relate to approval by, for example, the FDA, EMA, or MHRA. "Standard of care" second-line treatments for gastroesophageal cancer are well known in the art and are described in publicly available clinical guidelines, such as those provided by the National Comprehensive Cancer Network.

[0045] A second-line "standard of care" for gastroesophageal cancer may include, for example, chemoradiotherapy, surgery, chemotherapy, or other anti-cancer systemic therapy if the first-line "standard of care" included esophagectomy without chemoradiotherapy. A second-line "standard of care" for gastroesophageal cancer may include, for example, surgery (if the cancer is operable) or other anti-cancer systemic therapy if the first-line "standard of care" included chemoradiotherapy without surgery. A second-line "standard of care" for gastroesophageal cancer may include, for example, systemic anti-cancer therapy if metastatic gastroesophageal cancer develops after the first-line "standard of care."

[0046] The gastroesophageal cancer treated by the methods of the present disclosure may be, for example, (A) a gastroesophageal cancer that has recurred after curative-intent treatment for locally advanced cancer, or (B) a gastroesophageal cancer that has been initially diagnosed as unresectable locally advanced or metastatic cancer. Curative-intent treatments are described in more detail below. However, by way of example, a curative-intent treatment may be a treatment that has already been described as an approved or "standard of care" curative-intent treatment for one or more gastroesophageal cancers. In this context, approval may relate to approval by, for example, the FDA, EMA, or MHRA. "Standard of care" curative-intent treatments for gastroesophageal cancer are well known in the art and are described in publicly available clinical guidelines, such as those provided by the National Comprehensive Cancer Network.

[0047] Thus, the individual may be a gastroesophageal cancer patient who has been treated for gastroesophageal cancer. The individual may be a gastroesophageal cancer patient who has been treated with curative intent for gastroesophageal cancer. The individual may be a gastroesophageal cancer patient who has been treated with first-line gastroesophageal cancer treatment. The individual may be a gastroesophageal cancer patient who has been treated with first-line and second-line gastroesophageal cancer treatment. Such cancer patients form a subset of gastroesophageal cancer patients that is well recognized in the art. First-line and second-line treatments with curative intent are described in more detail below.

[0048] Gastroesophageal cancer may be prone to recurrence. For example, gastroesophageal cancer may be prone to recurrence, local invasion, and / or metastasis. The methods of the present disclosure may be aimed at treating recurrence, recurrence, local invasion, and / or metastasis.

[0049] Treatment with curative intent Curative treatment can refer to any treatment that has the potential to cure an individual of cancer. Curative treatment can refer, for example, to a therapy administered to an individual to attempt to cure cancer. Curative treatment can be administered before "selecting" treatment for relapsed or spread cancer.

[0050] As explained above, known treatments for gastroesophageal cancer include surgical resection, radiation therapy, and systemic therapy. A treatment with curative intent (such as a first curative intent treatment or a second curative intent treatment) may include (i) surgical resection, (ii) radiation therapy, and / or (iii) systemic therapy. For example, a treatment with curative intent may include (i); (ii); (iii); (i) and (ii); (i) and (iii); (ii) and (iii); or (i), (ii), and (iii). In certain embodiments of the present disclosure, a treatment with curative intent may include (i); (ii); or (i) and (ii).

[0051] Systemic therapy may include or consist of (a) chemotherapy. Systemic therapy may include or consist of (b) immunotherapy. Systemic therapy may include or consist of (c) targeted therapy. For example, systemic therapy may include or consist of (a); (b); (c); (a) and (b); (a) and (c); (b) and (c); or (a), (b), and (c).

[0052] Chemotherapy, such as that for gastroesophageal cancer, is well known in the art. Such chemotherapy may include, for example, platinum-based antineoplastic agents such as cisplatin, oxaliplatin, or carboplatin. Such chemotherapy may include, for example, antimetabolites such as fluorouracil (5-FU), trifluridine, or capecitabine. Such chemotherapy may include, for example, taxanes such as docetaxel or paclitaxel. Leucovorin may be administered in conjunction with chemotherapy to reduce the toxic effects of chemotherapy.

[0053] Immunotherapies, such as those for gastroesophageal cancer, are well known in the art. Such immunotherapies may include, for example, therapeutic immune cells, immunomodulators, checkpoint inhibitors, and vaccines. Therapeutic immune cells may include T cells, such as genetically engineered T cells, such as CAR T cells or T cells expressing a genetically engineered TCR. Immunomodulators may include, for example, interleukins, cytokines, chemokines, and immunomodulatory drugs. Checkpoint inhibitors may include, for example, CTLA-4 inhibitors or PD-1 axis binding antagonists. CTLA-4 inhibitors may include ipilimumab. PD-1 axis binding antagonists may include, for example, pembrolizumab, dostallimab-gxly, and nivolumab. Checkpoint inhibitors and PD-1 axis binding antagonists are described in more detail below.

[0054] Targeted therapy, such as that for gastroesophageal cancer, is well known in the art. The term targeted therapy is a term used in the art to refer to a treatment that targets specific genes and proteins that help cancer cells survive and grow. Targeted therapy can include, for example, tropomycin kinase receptor antagonists (such as larotrectinib or entrectinib), HER2 antagonists (such as trastuzumab), topoisomerase I inhibitors (such as irinotecan or deruxtecan-nxki), VEGF inhibitors (such as ramucirumab), or thymidine phosphorylase inhibitors (such as tiparisil).

[0055] Systemic therapy may include, for example, a drug (or drug combination) already described for the treatment of gastroesophageal cancer. Systemic therapy may include a drug or drug combination already described as an approved "standard of care" curative treatment for gastroesophageal cancer.

[0056] For example, systemic therapy may include a drug or drug combination already described for neoadjuvant chemoradiotherapy or chemotherapy in the "standard of care" curative treatment of operable gastroesophageal cancer (or gastroesophageal cancer for which local therapy is indicated). Systemic therapy may include, for example, paclitaxel, carboplatin, fluorouracil, oxaliplatin, cisplatin, irinotecan, and / or capecitabine. Systemic therapy may include, for example, combination therapy with paclitaxel and carboplatin; fluorouracil and oxaliplatin; fluorouracil and cisplatin; irinotecan and cisplatin; or paclitaxel and a fluoropyrimidine (fluorouracil or capecitabine).

[0057] Systemic therapy may include, for example, a drug or drug combination already described for perioperative chemotherapy in the "standard of care" curative treatment of operable gastroesophageal cancer (or gastroesophageal cancer for which local therapy is indicated). The gastroesophageal cancer may be, for example, adenocarcinoma of the thoracic esophagus or EGJ. Systemic therapy may include, for example, fluorouracil, leucovorin, oxaliplatin, docetaxel, fluoropyrimidines, and / or cisplatin. Systemic therapy may include, for example, combination therapy with fluorouracil, leucovorin, oxaliplatin, and docetaxel; fluoropyrimidines and oxaliplatin; or fluorouracil and cisplatin.

[0058] Systemic therapy may include, for example, drugs or drug combinations already described for definitive chemoradiotherapy in the "standard of care" cure-intent treatment for operable gastroesophageal cancer (or gastroesophageal cancer for which local therapy is indicated). Systemic therapy may include, for example, paclitaxel, carboplatin, fluorouracil, oxaliplatin, cisplatin, docetaxel, irinotecan, and / or fluoropyrimidines (fluorouracil or capecitabine). Systemic therapy may include, for example, combination therapy with paclitaxel and carboplatin; fluorouracil and oxaliplatin; fluorouracil and cisplatin; cisplatin and docetaxel or paclitaxel; irinotecan and cisplatin; or paclitaxel and fluoropyrimidines (fluorouracil or capecitabine).

[0059] Systemic therapy may include, for example, a drug or drug combination already described for postoperative therapy in the "standard of care" curative-intent treatment for operable gastroesophageal cancer (or gastroesophageal cancer for which local therapy is indicated). Postoperative therapy may include, for example, nivolumab, capecitabine, oxaliplatin, or fluorouracil. Systemic therapy may include, for example, monotherapy with nivolumab after postoperative therapy involving resection. Systemic therapy may include, for example, combination therapy with capecitabine and oxaliplatin; or fluorouracil and oxaliplatin.

[0060] Systemic therapy may include, for example, a drug or drug combination already described for postoperative chemoradiotherapy in the "standard of care" curative-intent treatment of operable gastroesophageal cancer (or gastroesophageal cancer for which local therapy is indicated). Systemic therapy may include, for example, a fluoropyrimidine (i.e., fluorouracil or capecitabine). Systemic therapy may include, for example, monotherapy with fluorouracil or capecitabine, before and / or after fluoropyrimidine-based chemoradiotherapy.

[0061] In either case, the selection of systemic therapy to include in the curative treatment can be informed by the characteristics of the cancer, such as the expression of specific markers. For example, if the cancer overexpresses HER2 (i.e., the cancer is HER2-overexpression-positive, e.g., HER2-overexpression-positive adenocarcinoma), the curative treatment may include trastuzumab (e.g., in combination with a fluoropyrimidine, oxaliplatin, or cisplatin, and optionally pembrolizumab). HER2 overexpression can be determined by any means known to those skilled in the art, such as immunohistochemistry (IHC), fluorescence in situ hybridization (FISH), other in situ hybridization (ISH), or next-generation sequencing (NGS). In surgical specimens, a cancer may be considered HER2-overexpression-positive if there is strong, complete, basolateral or outer membrane reactivity in ≥10% of the cancer cells. In biopsy specimens, a cancer may be considered HER2-overexpression positive if there are clusters of five or more cancer cells with strong, complete, basolateral, or outer membrane reactivity, regardless of the percentage of cancer cells present. If reactivity is weak to moderately complete, HER2 overexpression is equivocal.

[0062] If the cancer overexpresses PD-L1, treatment with curative intent may include a PD-1 axis-binding antagonist, such as nivolumab or pembrolizumab. PD-L1 testing can be used to determine whether an individual is a candidate for treatment with such an antagonist. Methods for determining PD-L1 expression are known to those of skill in the art. A cancer may be considered suitable for treatment with a PD-1 axis-binding inhibitor if at least 1%, e.g., at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, or at least 80% of gastroesophageal cancer cells from an individual express PD-L1. The percentage of cells expressing PD-L1 can be determined by any means known to those of skill in the art, such as immunohistochemistry (IHC), flow cytometry, or enzyme-linked immunosorbent assay (ELISA). A cancer may be considered to express PD-L1 if its CPS (combined positive score) is ≥ 1, e.g., ≥ 5 or ≥ 10.

[0063] If the cancer is an NTRK gene fusion-positive solid tumor, the curative treatment may include a TRK inhibitor, such as larotrectinib or entrectinib. The tumor can be identified by any means known to those skilled in the art, such as next-generation sequencing (NGS). Those skilled in the art can use NGS to determine tumors with high mutational burden (TMB).

[0064] In one aspect of the present disclosure, the curative treatment does not include therapeutic T cells, and thus the individual may not have received therapeutic T cells prior to the method of the present disclosure.

[0065] First- and second-line treatment The first-line and / or second-line treatments may be any known or unknown treatments for gastroesophageal cancer. As described above, known treatments for gastroesophageal cancer include surgical resection, radiation therapy, and systemic therapy. Thus, a treatment with curative intent (such as a first curative intent treatment or a second curative intent treatment) may include (i) surgical resection, (ii) radiation therapy, and / or (iii) systemic therapy. For example, a treatment with curative intent may include (i); (ii); (iii); (i) and (ii); (i) and (iii); (ii) and (iii); or (i), (ii), and (iii). In certain embodiments of the present disclosure, a treatment with curative intent may include (i); (ii); or (i) and (ii).

[0066] Systemic therapy may include or consist of (a) chemotherapy. Systemic therapy may include or consist of (b) immunotherapy. Systemic therapy may include or consist of (c) targeted therapy. For example, systemic therapy may include or consist of (a); (b); (c); (a) and (b); (a) and (c); (b) and (c); or (a), (b), and (c).

[0067] Chemotherapy, such as that for gastroesophageal cancer, is well known in the art. Such chemotherapy may include, for example, platinum-based antineoplastic agents such as cisplatin, oxaliplatin, or carboplatin. Such chemotherapy may include, for example, antimetabolites such as fluorouracil (5-FU), trifluridine, or capecitabine. Such chemotherapy may include, for example, taxanes such as docetaxel or paclitaxel. Leucovorin may be administered in conjunction with chemotherapy to reduce the toxic effects of chemotherapy.

[0068] Immunotherapies, such as those for gastroesophageal cancer, are well known in the art. Such immunotherapies may include, for example, therapeutic immune cells, immunomodulators, checkpoint inhibitors, and vaccines. Therapeutic immune cells may include T cells, such as genetically engineered T cells, such as CAR T cells or T cells expressing a genetically engineered TCR. Immunomodulators may include, for example, interleukins, cytokines, chemokines, and immunomodulatory drugs. Checkpoint inhibitors may include, for example, CTLA-4 inhibitors or PD-1 axis binding antagonists. CTLA-4 inhibitors may include ipilimumab. PD-1 axis binding antagonists may include, for example, pembrolizumab, dostallimab-gxly, and nivolumab. Checkpoint inhibitors and PD-1 axis binding antagonists are described in more detail below.

[0069] Targeted therapy, such as that for gastroesophageal cancer, is well known in the art. The term targeted therapy is a term used in the art to refer to a treatment that targets specific genes and proteins that help cancer cells survive and grow. Targeted therapy can include, for example, tropomycin kinase receptor antagonists (such as larotrectinib or entrectinib), HER2 antagonists (such as trastuzumab), topoisomerase I inhibitors (such as irinotecan or deruxtecan-nxki), VEGF inhibitors (such as ramucirumab), or thymidine phosphorylase inhibitors (such as tiparisil).

[0070] Systemic therapy can include, for example, a drug (or drug combination) already described for the treatment of gastroesophageal cancer. Systemic therapy can include a drug or drug combination already described as a first-line approved treatment or "standard of care" for gastroesophageal cancer.

[0071] For example, the systemic therapy may include a drug (or combination of drugs) already described for first-line treatment of the "standard of care" for gastroesophageal cancer, such as inoperable gastroesophageal cancer (or gastroesophageal cancer for which a surgical approach is less favorable). The systemic therapy may include, for example, a fluoropyrimidine (fluorouracil or capecitabine), oxaliplatin, trastuzumab, cisplatin, nivolumab, pembrolizumab, ipilimumab, irinotecan, paclitaxel, carboplatin, and / or docetaxel.Systemic therapy includes, for example, fluoropyrimidines (fluorouracil or capecitabine) and oxaliplatin and trastuzumab (e.g., for HER2-overexpressing adenocarcinoma); fluoropyrimidines (fluorouracil or capecitabine) and cisplatin and trastuzumab (e.g., for HER2-overexpressing adenocarcinoma); for adenocarcinoma (e.g., for HER2-overexpressing adenocarcinoma), fluoropyrimidines (fluorouracil or capecitabine), oxaliplatin, and nivolumab; fluoropyrimidines (fluorouracil or capecitabine) fluoropyrimidines (fluorouracil or capecitabine), oxaliplatin, and nivolumab (e.g., for HER2-overexpression-negative squamous cell carcinoma); fluoropyrimidines (fluorouracil or capecitabine), cisplatin, and nivolumab (e.g., for HER2-overexpression-negative squamous cell carcinoma); fluoropyrimidines (fluorouracil or capecitabine), oxaliplatin, and pembrolizumab (e.g., for HER2-overexpression-negative cancers); fluoropyrimidines (fluorouracil or capecitabine), cisplatin, and pembrolizumab (e.g., for HER2-overexpression-negative cancers) fluoropyrimidines (fluorouracil or capecitabine) and oxaliplatin (e.g., for HER2-negative cancers); fluoropyrimidines (fluorouracil or capecitabine) and cisplatin (e.g., for HER2-negative cancers); nivolumab and ipilimumab (e.g., for HER2-negative squamous cell carcinoma); fluoropyrimidines (fluorouracil or capecitabine) and cisplatin and trastuzumab and pembrolizumab (e.g., for HER2-positive adenocarcinoma); fluoropyrimidines docetaxel with or without cisplatin; fluoropyrimidines (fluorouracil or capecitabine); docetaxel, cisplatin or oxaliplatin, and fluorouracil; or combination therapy with docetaxel, carboplatin, and fluorouracil.

[0072] Systemic therapy can include, for example, a drug (or combination of drugs) already described for approved treatment or "standard of care" third-line or subsequent-line therapy for gastroesophageal cancer, such as inoperable gastroesophageal cancer (or gastroesophageal cancer for which a surgical approach is less favorable). Systemic therapy can include, for example, dostallimab-gxly, nivolumab, pembrolizumab, docetaxel, paclitaxel, irinotecan, entrectinib, larotrectinib, ramucirumab, fam-trastuzumab, deruxtecan-nxki, fluorouracil, and / or cisplatin. Systemic therapy can include monotherapy with, for example, dostallimab-gxly (e.g., for MSI-H or dMMR tumors), nivolumab (e.g., for esophageal squamous cell carcinoma), pembrolizumab (e.g., for MSI-H or dMMR tumors, or for high TMB tumors (≧10 mutations / megabase), or for second-line treatment of esophageal squamous cell carcinoma with PD-L1 expression levels of ≧10 by CPS), docetaxel, paclitaxel, irinotecan, entrectinib (e.g., for NTRK gene fusion-positive tumors), or larotrectinib (e.g., for NTRK gene fusion-positive tumors). Systemic therapy may include, for example, combination therapy with ramucirumab and paclitaxel (e.g., for adenocarcinoma such as EGJ adenocarcinoma or esophageal adenocarcinoma); fam-trastuzumab deruxtecan-nxki (e.g., for HER2-overexpressing adenocarcinoma); fluorouracil and irinotecan; ramucirumab for adenocarcinoma (e.g., for EGJ adenocarcinoma or esophageal adenocarcinoma); irinotecan and cisplatin; fluorouracil, irinotecan and ramucirumab (e.g., for adenocarcinoma); irinotecan and ramucirumab (e.g., for adenocarcinoma); or docetaxel and irinotecan.

[0073] Systemic therapy may include, for example, a drug (or drug combination) already described for approved treatment or second-line or subsequent-line therapy for "standard of care" gastroesophageal cancer, such as inoperable gastroesophageal cancer (or gastroesophageal cancer for which a surgical approach is less favorable). Systemic therapy may include, for example, trifluridine and / or tipiracil. Systemic therapy may include, for example, combination therapy with trifluridine and tipiracil (e.g., in the case of EGJ adenocarcinoma).

[0074] In either case, the choice of systemic therapy to include in treatment (e.g., first-line or second-line treatment) can be informed by the characteristics of the cancer, such as the expression of certain markers. For example, if the cancer overexpresses HER2 (i.e., the cancer is HER2-overexpression-positive, e.g., HER2-overexpression-positive adenocarcinoma), the first-line and / or second-line treatment may include trastuzumab (e.g., in combination with a fluoropyrimidine, oxaliplatin, or cisplatin, and optionally pembrolizumab). HER2 overexpression can be determined by any means known to those skilled in the art, such as immunohistochemistry (IHC), fluorescence in situ hybridization (FISH), other in situ hybridization (ISH), or next-generation sequencing (NGS). In surgical specimens, a cancer may be considered HER2-overexpression-positive if there is strong, complete, basolateral or outer membrane reactivity in ≥10% of the cancer cells. In biopsy specimens, a cancer may be considered HER2-overexpression positive if there are clusters of five or more cancer cells with strong, complete, basolateral, or outer membrane reactivity, regardless of the percentage of cancer cells present. If reactivity is weak to moderately complete, HER2 overexpression is equivocal.

[0075] If the cancer overexpresses PD-L1, first-line and / or second-line treatment may include a PD-1 axis-binding antagonist, such as nivolumab or pembrolizumab. PD-L1 testing can be used to determine whether an individual is a candidate for treatment with such an antagonist. Methods for determining PD-L1 expression are known to those of skill in the art. A cancer may be considered suitable for treatment with a PD-1 axis-binding inhibitor if at least 1%, e.g., at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, or at least 80% of the gastroesophageal cancer cells from the individual express PD-L1. The percentage of cells expressing PD-L1 can be determined by any means known to those of skill in the art, such as immunohistochemistry (IHC), flow cytometry, or enzyme-linked immunosorbent assay (ELISA). A cancer may be considered to express PD-L1 if its CPS (combined positive score) is ≧1, for example ≧5 or ≧10.

[0076] If the cancer is an NTRK gene fusion-positive solid tumor, the first-line and / or second-line treatment may include a TRK inhibitor, such as larotrectinib or entrectinib. The tumor can be identified by any means known to those skilled in the art, such as next-generation sequencing (NGS). Those skilled in the art can use NGS to determine tumors with high mutational burden (TMB).

[0077] In one embodiment of the present disclosure, the first-line and / or second-line treatment does not include therapeutic T cells, and thus the individual may not have received therapeutic T cells prior to the methods of the present disclosure.

[0078] Population of modified T cells The method involves administering to the individual a population of engineered T cells comprising a heterologous CD8 co-receptor and a heterologous TCR capable of binding to a peptide antigen of MAGE-A4. It is the presence of the heterologous CD8 co-receptor and the heterologous TCR that "engineer" the T cells. The heterologous CD8 co-receptor and the heterologous TCR are typically found on the surface of the engineered T cells. In other words, the engineered T cells can express the heterologous CD8 co-receptor and the heterologous TCR on their surface.

[0079] In the context of the present disclosure, the term "heterologous" refers to a polypeptide or nucleic acid that is foreign to a particular biological system (such as a T cell), i.e., that does not naturally occur in that system. A "heterologous" polypeptide or nucleic acid can be introduced into a system by artificial or recombinant means. Thus, heterologous expression of a TCR can alter the specificity of a T cell. Heterologous expression of a CD8 co-receptor can confer CD8 co-receptor-associated functions to a T cell. Heterologous CD8 co-receptors and heterologous TCRs are described in more detail below.

[0080] The modified T cells can include CD4+ T cells. That is, the modified T cells can include T cells that express an endogenous CD4 co-receptor. The modified T cells can include CD8+ T cells. That is, the modified T cells can include T cells that express an endogenous CD8 co-receptor. The modified T cells can include CD4+ T cells and CD8+ T cells. That is, the modified T cells can include T cells that express an endogenous CD4 co-receptor and can include T cells that express an endogenous CD8 co-receptor. Both the CD4+ T cells and the CD8+ T cells can carry a heterologous CD8 co-receptor.

[0081] The modified T cells may be allogeneic to the individual. The modified T cells may preferably be autologous to the individual. In this case, the modified T cells can be generated by modifying endogenous cells obtained from the individual. Thus, the method can include generating a population. Methods for generating modified T cells are known in the art and are discussed in the Examples below. Typically, the modified T cells of the present disclosure are generated from cells such as peripheral blood mononuclear cells (PBMCs). T cells are typically sorted from harvested cells and engineered to contain the desired modifications (here, a heterologous CD8 co-receptor and a heterologous TCR). Thus, the method can include (a) obtaining peripheral blood mononuclear cells (PBMCs) from the individual; (b) sorting T cells from the PBMCs; and (c) generating a population by modifying the selected T cells to express a heterologous CD8 co-receptor and a heterologous TCR.

[0082] Autologous modified T cells may be generated in anticipation of an individual's need. That is, the autologous modified T cells may be generated in advance, before the individual requires treatment with the modified cells. This can help ensure that autologous modified T cells are available for administration as soon as possible after it is determined that the individual requires treatment. In this way, the individual does not have to wait for autologous T cells to be generated before treatment can begin. This can improve the outcome of the treatment.

[0083] The proactive generation of autologous T cells may be particularly relevant in the treatment of gastroesophageal cancer patients who are at high risk of recurrence and / or who are at high risk of failure of first-line and / or second-line "standard of care." The risk of recurrence for individuals who have received "standard of care" for gastroesophageal cancer can be determined by methods routinely performed in the art. Such methods may include monitoring clinical signs or symptoms. Such methods may include, for example, one or more magnetic resonance imaging (MRI), positron emission tomography (PET), and / or computed tomography (CT) scans performed after treatment to monitor tumor progression and / or recurrence. MRI, PET, and / or CT scans may be performed, for example, approximately every three months (e.g., once every four to sixteen weeks or once every eight to twelve weeks). If an individual is identified as being at high risk of recurrence, PBMCs may be obtained at this time (i.e., before recurrence) for the purpose of generating autologous modified T cells ready for administration at the time of recurrence. Thus, when the cancer is recurrent gastroesophageal cancer, the method of the present disclosure may include: (a) obtaining peripheral blood mononuclear cells (PBMCs) from the individual; (b) sorting T cells from the PBMCs; and (c) generating a population by modifying the sorted T cells to express a heterologous CD8 co-receptor and a heterologous TCR, wherein one or more of steps (a)-(c) are performed before recurrence. Preferably, step (a) is performed before recurrence. More preferably, steps (a) and (b) are performed before recurrence. Most preferably, steps (a), (b), and (c) are performed before recurrence. Any of these options are effective in providing treatment one step ahead of recurrence.

[0084] In either case, the population of modified T cells may be administered, for example, as a single dose. The population of modified T cells may be administered, for example, as a single dose, as soon as possible after a diagnosis of gastroesophageal cancer. For example, the population of modified T cells may be administered as soon as possible after a recurrence of gastroesophageal cancer is confirmed. For example, the population of modified T cells may be administered as soon as possible after a diagnosis of previously untreated gastroesophageal cancer is confirmed.

[0085] In the context of the present disclosure, the term "as soon as possible" can refer to the earliest time point at which it is practical to administer the population of modified T cells. As explained above, generation of autologous modified T cells can take some time, so there may be a time lag between the diagnosis of gastroesophageal cancer (e.g., recurrent gastroesophageal cancer) and the administration of treatment. Thus, administering as soon as possible after diagnosis can refer to administering the autologous modified T cells as soon as practical after they are generated. Administration as soon as possible after diagnosis can refer, for example, to administration from less than about 150 days after diagnosis of gastroesophageal cancer (e.g., recurrent gastroesophageal cancer), e.g., less than about 125 days, less than about 100 days, less than about 90 days, less than about 80 days, less than about 70 days, less than about 60 days, less than about 50 days, less than about 40 days, or less than about 30 days after diagnosis of gastroesophageal cancer (e.g., recurrent gastroesophageal cancer). The population can be administered to an individual, for example, about 30 to about 150 days after diagnosis of gastroesophageal cancer (e.g., recurrent gastroesophageal cancer), for example, about 40 to about 125 days, about 50 to about 100 days, about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, about 65 days, or about 60 days after diagnosis of gastroesophageal cancer (e.g., recurrent gastroesophageal cancer).

[0086] Heterogeneous TCR The engineered T cells comprise a heterologous TCR capable of binding to a peptide antigen of MAGE-A4. In other words, the engineered T cells express or display, for example, on their surface, a heterologous TCR capable of binding to a peptide antigen of MAGE-A4. MAGE-A4 is a well-known cancer antigen whose expression is restricted in normal (i.e., non-cancerous) tissues. MAGE-A4 has been shown to suppress p53 targets (such as BAX and CDKN1A) and is a binding partner of the oncogene gankyrin.

[0087] The heterologous TCR is capable of binding to a peptide antigen of MAGE-A4. For example, the heterologous TCR can bind to GVYDGREHTV (SEQ ID NO: 1), a peptide sequence known as MAGE-A4 230-239 contained in MAGE-A4. The heterologous TCR can bind to, for example, a peptide antigen of MAGE-A4 (e.g., GVYDGREHTV (SEQ ID NO: 1)) and an HLA-A molecule (e.g., HLA-A *02 or HLA-A * A heterologous TCR can bind to a complex containing an HLA molecule, such as an HLA molecule (HLA-0201 molecule). In either case, the binding can be specific. Specificity refers to the strength of binding between a heterologous TCR and its target antigen. Specificity can be expressed by the dissociation constant Kd, which is the ratio of the bound state to the unbound state of the receptor-ligand system. Typically, the fewer different antigens a heterologous TCR can bind to other than MAGE-A4, the higher its binding specificity.

[0088] The xenogeneic TCR can bind, for example, to a MAGE-A4 peptide antigen (e.g., SEQ ID NO: 1) or a complex comprising a MAGE-A4 peptide antigen (e.g., SEQ ID NO: 1) and an HLA molecule with a dissociation constant (Kd) of 0.01 μM to 100 μM, 0.01 μM to 50 μM, 0.01 μM to 20 μM, 10 μM to 1000 μM, 10 μM to 500 μM, or 50 μM to 500 μM. For example, in a preferred embodiment of the present disclosure, the xenogeneic TCR binds to a MAGE-A4 peptide antigen or a complex comprising a MAGE-A4 peptide antigen and an HLA molecule with a Kd of 0.05 μM to 20.0 μM. For example, a heterologous TCR may be expressed at 0.01 μM, 0.02 μM, 0.03 μM, 0.04 μM, 0.05 μM, 0.06 μM, 0.07 μM, 0.08 μM, 0.09 μM, 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, 0.8 μM, 0.85 μM, 0.9 μM, 0.95μΜ, 1.0μΜ, 1.5μΜ, 2.0μΜ, 2.5μΜ, 3.0μΜ, 3.5μΜ, 4.0μΜ, 4.5μΜ, 5. 0μΜ, 5.5μΜ, 6.0μΜ, 6.5μΜ, 7.0μΜ, 7.5μΜ, 8.0μΜ, 8.5μΜ, 9.0μΜ, 9.5μΜ, The TCR can bind with a Kd of 10.0 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, 100 μM, 150 μM, 200 μM, 250 μM, 300 μM, 350 μM, 400 μM, 450 μM, or 500 μM. The Kd can be measured, for example, using surface plasmon resonance, optionally at 25° C., optionally at pH 6.5-6.9 or 7.0-7.5. The dissociation constant Kd or koff / kon can be determined by experimentally measuring the dissociation rate constant koff and the association rate constant kon. The TCR dissociation constant can be measured using a soluble form of the TCR, wherein the TCR comprises a TCR alpha chain variable domain and a TCR beta chain variable domain.

[0089] The heterologous TCR may be, for example, a recombinant, synthetic, or artificial TCR. That is, the heterologous TCR may be a TCR that does not occur in nature. The heterologous TCR may be, for example, an affinity-enhanced TCR, such as a specific peptide-enhanced affinity receptor (SPEAR™) TCR.

[0090] The heterologous TCR may, for example, comprise an alpha chain variable domain having at least 80% sequence identity to the sequence of amino acid residues 22-125 of SEQ ID NO: 2. The heterologous TCR may, for example, comprise a beta chain variable domain having at least 80% sequence identity to the sequence of amino acid residues 22-123 of SEQ ID NO: 3. The heterologous TCR may, for example, comprise an alpha chain variable domain having at least 80% sequence identity to the sequence of amino acid residues 22-125 of SEQ ID NO: 2 and a beta chain variable domain having at least 80% sequence identity to the sequence of amino acid residues 22-123 of SEQ ID NO: 3. The alpha chain variable domain may, for example, have at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence of amino acid residues 22-125 of SEQ ID NO: 2. The alpha chain variable domain may, for example, comprise or consist of amino acid residues 22-125 of SEQ ID NO: 2. The beta chain variable domain can have, for example, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence of amino acid residues 22 to 123 of SEQ ID NO: 3. The beta chain amino acid sequence can, for example, comprise or consist of amino acid residues 22 to 123 of SEQ ID NO: 3.

[0091] The heterologous TCR may, for example, comprise an alpha chain having at least 80% sequence identity to the sequence of amino acid residues 22-282 of SEQ ID NO: 2. The heterologous TCR may, for example, comprise a beta chain having at least 80% sequence identity to the sequence of amino acid residues 22-311 of SEQ ID NO: 3. The heterologous TCR may, for example, comprise an alpha chain having at least 80% sequence identity to the sequence of amino acid residues 22-282 of SEQ ID NO: 2 and a beta chain variable domain having at least 80% sequence identity to the sequence of amino acid residues 22-311 of SEQ ID NO: 3. The alpha chain variable domain may, for example, have at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence of amino acid residues 22-282 of SEQ ID NO: 2. The alpha chain variable domain may, for example, comprise or consist of amino acid residues 22-282 of SEQ ID NO: 2. The beta chain variable domain can have, for example, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence of amino acid residues 22 to 311 of SEQ ID NO: 3. The beta chain amino acid sequence can, for example, comprise or consist of amino acid residues 22 to 311 of SEQ ID NO: 3.

[0092] Heterologous TCRs are typically expressed with an N-terminal signal peptide that is cleaved prior to expression at the surface of a T cell. In this regard, amino acids 1-21 of SEQ ID NO:2 and SEQ ID NO:3, respectively, are typically cleaved prior to expression of the TCR at the surface of a T cell. A heterologous TCR may, for example, comprise an alpha chain amino acid sequence having at least 80% sequence identity to SEQ ID NO:2. A heterologous TCR may, for example, comprise a beta chain amino acid sequence having at least 80% sequence identity to SEQ ID NO:3. A heterologous TCR may, for example, comprise an alpha chain amino acid sequence having at least 80% sequence identity to SEQ ID NO:2 and a beta chain amino acid sequence having at least 80% sequence identity to SEQ ID NO:3. The alpha chain amino acid sequence may, for example, have at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:2. The alpha chain amino acid sequence may, for example, comprise or consist of SEQ ID NO:2. The beta chain amino acid sequence can have, for example, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 3. The beta chain amino acid sequence can, for example, comprise or consist of SEQ ID NO: 3.

[0093] Heterologous TCRs can be, for example, (i) an alpha chain variable domain comprising a CDR1 comprising (1) the sequence of SEQ ID NO:4 or (2) an amino acid sequence comprising an insertion, deletion, or substitution of one, two, or three amino acids relative to the sequence of SEQ ID NO:4; (ii) an alpha chain variable domain comprising a CDR2 comprising (1) the sequence of SEQ ID NO:5 or (2) an amino acid sequence comprising an insertion, deletion, or substitution of one, two, or three amino acids relative to the sequence of SEQ ID NO:5; or (iii) an alpha chain variable domain comprising a CDR3 comprising (1) the sequence of SEQ ID NO:6 or (2) an amino acid sequence comprising an insertion, deletion, or substitution of one, two, or three amino acids relative to the sequence of SEQ ID NO:6. (iv) a beta chain variable domain comprising a CDR1 comprising (1) the sequence of SEQ ID NO:7 or (2) an amino acid sequence comprising an insertion, deletion, or substitution of one, two, or three amino acids relative to the sequence of SEQ ID NO:7; (v) a beta chain variable domain comprising a CDR2 comprising (1) the sequence of SEQ ID NO:8 or (2) an amino acid sequence comprising an insertion, deletion, or substitution of one, two, or three amino acids relative to the sequence of SEQ ID NO:8; and / or (vi) a beta chain variable domain comprising a CDR3 comprising (1) the sequence of SEQ ID NO:9 or (2) an amino acid sequence comprising an insertion, deletion, or substitution of one, two, or three amino acids relative to the sequence of SEQ ID NO:9.

[0094] The alpha chain of a heterologous TCR may comprise, for example, (i) an alpha chain variable domain comprising a CDR1 comprising (1) the sequence of SEQ ID NO: 4, or (2) an amino acid sequence comprising an insertion, deletion, or substitution of one, two, or three amino acids relative to the sequence of SEQ ID NO: 4; (ii) an alpha chain variable domain comprising a CDR2 comprising (1) the sequence of SEQ ID NO: 5, or (2) an amino acid sequence comprising an insertion, deletion, or substitution of one, two, or three amino acids relative to the sequence of SEQ ID NO: 5; and (iii) an alpha chain variable domain comprising a CDR3 comprising (1) the sequence of SEQ ID NO: 6, or (2) an amino acid sequence comprising an insertion, deletion, or substitution of one, two, or three amino acids relative to the sequence of SEQ ID NO: 6. The alpha chain of a heterologous TCR may comprise, for example, (i) an alpha chain variable domain comprising a CDR1 comprising the sequence of SEQ ID NO: 4; (ii) an alpha chain variable domain comprising a CDR2 comprising the sequence of SEQ ID NO: 5; and (iii) an alpha chain variable domain comprising a CDR3 comprising the sequence of SEQ ID NO: 6.

[0095] The beta chain of a heterologous TCR may comprise, for example, (vi) a beta chain variable domain comprising a CDR1 comprising (1) the sequence of SEQ ID NO: 7 or (2) an amino acid sequence comprising an insertion, deletion, or substitution of one, two, or three amino acids relative to the sequence of SEQ ID NO: 7; (v) a beta chain variable domain comprising a CDR2 comprising (1) the sequence of SEQ ID NO: 8 or (2) an amino acid sequence comprising an insertion, deletion, or substitution of one, two, or three amino acids relative to the sequence of SEQ ID NO: 8; and (vi) a beta chain variable domain comprising a CDR3 comprising (1) the sequence of SEQ ID NO: 9 or (2) an amino acid sequence comprising an insertion, deletion, or substitution of one, two, or three amino acids relative to the sequence of SEQ ID NO: 9. The beta chain of a heterologous TCR may comprise, for example, (iv) a beta chain variable domain comprising a CDR1 comprising the sequence of SEQ ID NO: 7; (v) a beta chain variable domain comprising a CDR2 comprising the sequence of SEQ ID NO: 8; and (vi) a beta chain variable domain comprising a CDR3 comprising the sequence of SEQ ID NO: 9.

[0096] The heterologous TCR may, for example, comprise an alpha chain comprising a CDR1 having the sequence of SEQ ID NO: 4, a CDR2 having the sequence of SEQ ID NO: 5, and a CDR3 having the sequence of SEQ ID NO: 6, and a beta chain comprising a CDR1 having the sequence of SEQ ID NO: 7, a CDR2 having the sequence of SEQ ID NO: 8, and a CDR3 having the sequence of SEQ ID NO: 9. The heterologous TCR may further comprise, for example, any of the percent identities of the alpha and beta chains discussed herein.

[0097] Heterologous CD8 co-receptor The modified T cells comprise a heterologous CD8 co-receptor, in other words, the modified T cells express, for example, a heterologous CD8 co-receptor on their surface.

[0098] CD8 is a cell surface glycoprotein naturally found on most cytotoxic T lymphocytes and mediates efficient cell-cell interactions within the immune system. CD8 acts as a co-receptor for the T cell receptor, so that CD8 and the T cell receptor together recognize antigens presented by antigen-presenting cells in the context of class I MHC molecules. The CD8 co-receptor binds to class I MHC and enhances TCR signaling. Functional co-receptors can be homodimers consisting of two CD8 alpha chains or heterodimers consisting of one CD8 alpha chain and one CD8 beta chain.

[0099] Thus, the heterologous CD8 co-receptor comprised in the engineered T cell may be CD8α. In other words, the heterologous CD8 co-receptor may be a homodimer consisting of two CD8 alpha chains. Alternatively, the heterologous CD8 co-receptor may be a heterodimer consisting of one CD8 alpha chain and one CD8 beta chain. In either case, the CD8 alpha chain may comprise or consist of an amino acid sequence having at least 80% (e.g., at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 10. Thus, the heterologous CD8 co-receptor may comprise an amino acid sequence having at least 80% (e.g., at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 10.

[0100] Both the CD8 alpha and beta chains share high homology with immunoglobulin variable light chains. The CD8 alpha and beta chains have CDR-like loops involved in MHC-class I binding. A heterologous CD8 coreceptor can comprise, for example, a CD8 alpha chain comprising: (i) an alpha chain CDR1 comprising (1) the sequence of SEQ ID NO: 11 or (2) an amino acid sequence containing one, two, or three amino acid insertions, deletions, or substitutions relative to the sequence of SEQ ID NO: 11; (ii) an alpha chain CDR2 comprising (1) the sequence of SEQ ID NO: 12 or (2) an amino acid sequence containing one, two, or three amino acid insertions, deletions, or substitutions relative to the sequence of SEQ ID NO: 12; and / or (iii) an alpha chain CDR3 comprising (1) the sequence of SEQ ID NO: 13 or (2) an amino acid sequence containing one, two, or three amino acid insertions, deletions, or substitutions relative to the sequence of SEQ ID NO: 13.

[0101] The heterologous CD8 co-receptor is capable of binding to a class I MHC molecule. The heterologous CD8 co-receptor can bind, for example, to the α3 portion of a class I MHC molecule, e.g., via an IgV-like domain of the CD8 co-receptor. The α3 portion is typically found between residues 223 and 229 of a class I MHC molecule. The ability of the heterologous CD8 co-receptor to bind to a class I MHC molecule improves the ability of the modified T cell to engage with a cognate antigen via the heterologous TCR. The cognate antigen, MAGE-A4, typically binds to HLA-A. * The heterologous CD8 co-receptor is presented as a complex with class I MHC molecules such as 02. The off-rate (k off ) can be improved or increased. This improvement or increase can be for engineered T cells that contain a heterologous TCR that binds to MAGE-A4 but lack a heterologous CD8 co-receptor. The heterologous CD8 co-receptor can, for example, help organize the heterologous TCR on the surface of the engineered cell, thereby improving the ability of the heterologous TCR to participate in TCR / peptide-MHCI interactions. The heterologous CD8 co-receptor can, for example, bind or interact with LCK (lymphocyte-specific protein tyrosine kinase) in a zinc-dependent manner and activate transcription factors such as NFAT, NF-κB, and AP-1. Thus, expression of a heterologous CD8 co-receptor can improve the affinity and / or avidity of the engineered T cells for MAGE-A4 and / or improve activation upon binding to MAGE-A4. Methods for determining affinity, avidity, and T cell activation are well known in the art. Expression of a heterologous CD8 co-receptor can improve or increase, for example, the expression of CD40L, cytokine production, cytotoxic activity, induction of dendritic cell maturation, or induction of dendritic cell cytokine production of the engineered T cells in response to antigen (MAGE-A4) binding, relative to engineered T cells that contain a heterologous TCR that binds to a peptide antigen of MAGE-A4 but lack a heterologous CD8 co-receptor.

[0102] CD8α and HLA-A *Synergy has been demonstrated between the heterologous CD8 co-receptor and the peptide antigen presented on HLA-A. Thus, in one embodiment of the present disclosure, the heterologous CD8 co-receptor may be CD8α and the heterologous TCR may be HLA-A. * It may be capable of binding to a peptide antigen of MAGE-A4 complexed with 0201. The peptide antigen may be, for example, SEQ ID NO:1.

[0103] Additional anticancer therapy The method may further comprise administering to the individual an additional anti-cancer therapy. That is, the method may comprise administering to the individual (i) a population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor (TCR) capable of binding to a peptide antigen of MAGE-A4, and (ii) an additional anti-cancer therapy. In other words, the method may comprise combination treatment with (i) a population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor (TCR) capable of binding to a peptide antigen of MAGE-A4, and (ii) an additional anti-cancer therapy. A checkpoint inhibitor, such as a PD-1 axis binding antagonist, may be included in the combination.

[0104] The individual may be administered at least one additional anti-cancer therapy, for example, one or more, two or more, three or more, four or more, or five or more additional anti-cancer therapies.

[0105] The additional anti-cancer therapy may be administered to the individual in the same selection of treatment as the population of modified T cells. As noted above, a "selection" of treatment may refer to a particular treatment regimen for a cancer that has failed a curative treatment or for which a curative treatment may be inappropriate. The additional anti-cancer therapy and population of modified T cells may be administered as part of a first-line treatment regimen. The additional anti-cancer therapy and population of modified T cells may be administered as part of a second-line treatment regimen. A second-line treatment regimen may be employed, for example, after failure of a first-line treatment regimen. The additional anti-cancer therapy and population of modified T cells may be administered as part of a third-line treatment regimen. A third-line treatment regimen may be employed, for example, after failure of a second-line treatment regimen.

[0106] The additional anti-cancer therapy may be, for example, an additional anti-cancer drug therapy. In other words, the additional anti-cancer therapy may be a systemic therapy. The additional anti-cancer therapy may, for example, include or consist of (a) chemotherapy. The additional anti-cancer therapy may, for example, include or consist of (b) immunotherapy. The additional anti-cancer therapy may, for example, include or consist of (c) targeted therapy. For example, the additional anti-cancer therapy may include or consist of (a); (b); (c); (a) and (b); (a) and (c); (b) and (c); or (a), (b), and (c).

[0107] Optional chemotherapy may be included in the additional anticancer therapy. The method may include administering one or more chemotherapeutic agents (e.g., two or more, three or more, four or more, or five or more chemotherapeutic agents). Chemotherapy, such as that for gastroesophageal cancer, is well known in the art. Such chemotherapy may include, for example, a platinum-based anti-neoplastic agent, such as cisplatin, oxaliplatin, or carboplatin. Such chemotherapy may include, for example, an antimetabolite, such as a fluoropyrimidine (e.g., fluorouracil or capecitabine), gemcitabine, trifluridine, or methotrexate. Such chemotherapy may include, for example, administering a taxane, such as docetaxel or paclitaxel. Such chemotherapy may include, for example, a topoisomerase I inhibitor, such as irinotecan or deruxtecan-nxki. Such chemotherapy may include, for example, a thymidine phosphorylase inhibitor, such as tipiracil. Leucovorin may be administered together with one or more chemotherapeutic agents to minimize toxic effects.

[0108] Any immunotherapy may be included in the additional anticancer therapy. The method may include administering one or more immunotherapies (e.g., two or more, three or more, four or more, or five or more immunotherapies). Immunotherapies, such as those for gastroesophageal cancer, are well known in the art. Such immunotherapies may include, for example, therapeutic immune cells, immunomodulators, checkpoint inhibitors, and vaccines. Therapeutic immune cells may include T cells, e.g., engineered T cells, such as CAR T cells or T cells expressing an engineered TCR. Immunomodulators may include, for example, interleukins, cytokines, chemokines, and immunomodulatory agents. Checkpoint inhibitors may include, for example, CTLA-4 inhibitors or PD-1 axis binding antagonists. CTLA-4 inhibitors may include ipilimumab. PD-1 axis binding antagonists may include, for example, pembrolizumab, dostallimab-gxly, and nivolumab. Checkpoint inhibitors and PD-1 axis binding antagonists are described in more detail below.

[0109] Any targeted therapy may be included in the additional anticancer therapy. The method may include administering one or more targeted therapies (e.g., two or more, three or more, four or more, or five or more targeted therapies). Targeted therapies, such as those for gastroesophageal cancer, are well known in the art. Such targeted therapies may include, for example, ramucirumab, a direct VEGFR2 antagonist. Such targeted therapies may include, for example, entrectinib, a selective tyrosine kinase inhibitor of tropomyosin receptor kinases A, B, and C, C-ros oncogene 1, and anaplastic lymphoma kinase. Such targeted therapies may include, for example, larotrectinib, an inhibitor of tropomyosin kinase receptors TrkA, TrkB, and TrkC. Such targeted therapies may include, for example, trastuzumab, a HER2-specific antibody. Other targeted therapies include bemarituzumab (FPA144), DKN-01, tebotelimab (MGD013), bactosertib (TEW-7197), and zolbetuximab (IMAB362).

[0110] The additional anti-cancer therapy may include a drug or drug combination already described for gastroesophageal cancer therapy. The additional anti-cancer therapy may include a drug or drug combination already described as a "standard of care" for gastroesophageal cancer. Thus, the method may include administering a drug or drug combination already described for gastroesophageal cancer therapy. The method may include administering a drug or drug combination already described as an approved "standard of care" for gastroesophageal cancer. In this context, approval may relate to approval by, for example, the FDA, EMA, or MHRA.

[0111] The additional anticancer therapy may include, for example, a drug (or drug combination) already described for use in preoperative chemoradiotherapy for gastroesophageal cancer. The additional anticancer therapy may include a drug or drug combination already described as a "standard of care" preoperative chemoradiotherapy for gastroesophageal cancer. The gastroesophageal cancer may be, for example, operable gastroesophageal cancer or gastroesophageal cancer for which local therapy is indicated. The additional anticancer therapy may include, for example, paclitaxel, carboplatin, fluorouracil, oxaliplatin, cisplatin, irinotecan, and / or capecitabine. The additional anticancer therapy may include, for example, combination therapy with paclitaxel and carboplatin; fluorouracil and oxaliplatin; fluorouracil and cisplatin; irinotecan and cisplatin; or paclitaxel and a fluoropyrimidine (fluorouracil or capecitabine).

[0112] The additional anticancer therapy may include, for example, a drug (or drug combination) already described for use in perioperative chemotherapy for gastroesophageal cancer. The additional anticancer therapy may include a drug or drug combination already described as a "standard of care" perioperative chemotherapy treatment for gastroesophageal cancer. The gastroesophageal cancer may be, for example, operable gastroesophageal cancer or gastroesophageal cancer for which local therapy is indicated. The gastroesophageal cancer may be, for example, adenocarcinoma of the thoracic esophagus at the EGJ. The additional anticancer therapy may include, for example, fluorouracil, leucovorin, oxaliplatin, docetaxel, fluoropyrimidines, and / or cisplatin. The additional anticancer therapy may include, for example, combination therapy with fluorouracil, leucovorin, oxaliplatin, and docetaxel; fluoropyrimidines and oxaliplatin; or fluorouracil and cisplatin.

[0113] The additional anti-cancer therapy may include, for example, a drug (or drug combination) already described for use in definitive chemoradiotherapy for gastroesophageal cancer. The additional anti-cancer therapy may include a drug or drug combination already described as a "standard of care" definitive chemoradiotherapy for gastroesophageal cancer. The gastroesophageal cancer may be, for example, operable gastroesophageal cancer or gastroesophageal cancer for which local therapy is indicated. The additional anti-cancer therapy may include, for example, paclitaxel, carboplatin, fluorouracil, oxaliplatin, cisplatin, docetaxel, irinotecan, and / or a fluoropyrimidine (fluorouracil or capecitabine). Additional anti-cancer therapies may include, for example, combination therapy with paclitaxel and carboplatin; fluorouracil and oxaliplatin; fluorouracil and cisplatin; cisplatin and docetaxel or paclitaxel; irinotecan and cisplatin; or paclitaxel and a fluoropyrimidine (fluorouracil or capecitabine).

[0114] The additional anticancer therapy may include, for example, a drug (or drug combination) already described for use in postoperative therapy for gastroesophageal cancer. The additional anticancer therapy may include a drug or drug combination already described as a "standard of care" postoperative therapy for gastroesophageal cancer. The gastroesophageal cancer may be, for example, operable gastroesophageal cancer or gastroesophageal cancer for which local therapy is indicated. The additional anticancer therapy may include, for example, nivolumab, capecitabine, oxaliplatin, or fluorouracil. The additional anticancer therapy may include, for example, postoperative therapy with resection followed by monotherapy with nivolumab. The additional anticancer therapy may include, for example, combination therapy with capecitabine and oxaliplatin; or fluorouracil and oxaliplatin.

[0115] The additional anti-cancer therapy may include, for example, a drug (or drug combination) already described for use in postoperative chemoradiotherapy for gastroesophageal cancer. The additional anti-cancer therapy may include a drug or drug combination already described as a "standard of care" postoperative chemoradiotherapy for gastroesophageal cancer. The gastroesophageal cancer may be, for example, operable gastroesophageal cancer or gastroesophageal cancer for which local therapy is indicated. The additional anti-cancer therapy may include, for example, a fluoropyrimidine (i.e., fluorouracil or capecitabine). The additional anti-cancer therapy may include, for example, monotherapy with fluorouracil or capecitabine before and / or after fluoropyrimidine-based chemoradiotherapy.

[0116] The additional anti-cancer therapy may include, for example, a drug (or drug combination) already described for first-line treatment of gastroesophageal cancer. The additional anti-cancer therapy may include a drug or drug combination already described as a "standard of care" first-line treatment for gastroesophageal cancer. The gastroesophageal cancer may be, for example, unresectable locally advanced, recurrent, or metastatic gastroesophageal cancer. Local therapy may not be indicated for unresectable locally advanced, recurrent, or metastatic gastroesophageal cancer. The additional anti-cancer therapy may include, for example, a fluoropyrimidine (fluorouracil or capecitabine), oxaliplatin, trastuzumab, cisplatin, nivolumab, pembrolizumab, ipilimumab, irinotecan, paclitaxel, carboplatin, and / or docetaxel.Additional anticancer therapies include, for example, fluoropyrimidines (fluorouracil or capecitabine) and oxaliplatin and trastuzumab (e.g., for HER2-overexpressing adenocarcinoma); fluoropyrimidines (fluorouracil or capecitabine) and cisplatin and trastuzumab (e.g., for HER2-overexpressing adenocarcinoma); for adenocarcinoma (e.g., for HER2-overexpressing adenocarcinoma), fluoropyrimidines (fluorouracil or capecitabine), oxaliplatin, and nivolumab; fluoropyrimidines (fluorouracil or capecitabine) and cisplatin and trastuzumab (e.g., for HER2-overexpressing adenocarcinoma). fluoropyrimidines (fluorouracil or capecitabine), oxaliplatin, and nivolumab (e.g., for HER2-overexpression-negative squamous cell carcinoma); fluoropyrimidines (fluorouracil or capecitabine), cisplatin, and nivolumab (e.g., for HER2-overexpression-negative squamous cell carcinoma); fluoropyrimidines (fluorouracil or capecitabine), oxaliplatin, and pembrolizumab (e.g., for HER2-overexpression-negative cancers); fluoropyrimidines (fluorouracil or capecitabine), cisplatin, and pembrolizumab (e.g., for HER2-overexpression-negative cancers) fluoropyrimidines (fluorouracil or capecitabine) and oxaliplatin (e.g., for HER2-negative cancers); fluoropyrimidines (fluorouracil or capecitabine) and cisplatin (e.g., for HER2-negative cancers); nivolumab and ipilimumab (e.g., for HER2-negative squamous cell carcinoma); fluoropyrimidines (fluorouracil or capecitabine) and cisplatin and trastuzumab and pembrolizumab (e.g., for HER2-positive adenocarcinoma); fluoropyrimidines combination therapy with fluoropyrimidines (fluorouracil or capecitabine) and oxaliplatin and trastuzumab and pembrolizumab (e.g., in the case of HER2-overexpressing adenocarcinoma); fluorouracil and irinotecan; paclitaxel with or without cisplatin or carboplatin; docetaxel with or without cisplatin; fluoropyrimidines (fluorouracil or capecitabine); docetaxel, cisplatin or oxaliplatin, and fluorouracil; or docetaxel, carboplatin, and fluorouracil.

[0117] The additional anticancer therapy may include, for example, a drug (or drug combination) already described for second-line or subsequent-line therapy for gastroesophageal cancer. The additional anticancer therapy may include a drug or drug combination already described as a second-line or subsequent-line treatment for "standard of care" for gastroesophageal cancer. The gastroesophageal cancer may be, for example, unresectable locally advanced, recurrent, or metastatic gastroesophageal cancer. Local therapy for unresectable locally advanced, recurrent, or metastatic gastroesophageal cancer may not be indicated. The additional anticancer therapy may include, for example, dostarlimab-gxly, nivolumab, pembrolizumab, docetaxel, paclitaxel, irinotecan, entrectinib, larotrectinib, ramucirumab, fam-trastuzumab, deruxtecan-nxki, fluorouracil, and / or cisplatin. Additional anticancer therapy may include monotherapy with, for example, dostallimab-gxly (e.g., for MSI-H or dMMR tumors), nivolumab (e.g., for esophageal squamous cell carcinoma), pembrolizumab (e.g., for MSI-H or dMMR tumors, or for high TMB tumors (≧10 mutations / megabase), or for second-line treatment of esophageal squamous cell carcinoma with PD-L1 expression levels of ≧10 by CPS), docetaxel, paclitaxel, irinotecan, entrectinib (e.g., for NTRK gene fusion-positive tumors), or larotrectinib (e.g., for NTRK gene fusion-positive tumors). Additional anti-cancer therapies may include, for example, combination therapy with ramucirumab and paclitaxel (e.g., in the case of adenocarcinoma such as EGJ adenocarcinoma or esophageal adenocarcinoma); fam-trastuzumab deruxtecan-nxki (e.g., in the case of HER2-overexpressing adenocarcinoma); fluorouracil and irinotecan; ramucirumab in the case of adenocarcinoma (e.g., in the case of EGJ adenocarcinoma or esophageal adenocarcinoma); irinotecan and cisplatin; fluorouracil, irinotecan and ramucirumab (e.g., in the case of adenocarcinoma); irinotecan and ramucirumab (e.g., in the case of adenocarcinoma); or docetaxel and irinotecan.

[0118] The additional anti-cancer therapy may include, for example, a drug (or drug combination) already described for third-line or subsequent-line therapy for gastroesophageal cancer. The additional anti-cancer therapy may include a drug or drug combination already described as a third-line or subsequent-line treatment of the "standard of care" for gastroesophageal cancer. The gastroesophageal cancer may be, for example, unresectable locally advanced, recurrent, or metastatic gastroesophageal cancer. Local therapy may not be indicated for unresectable locally advanced, recurrent, or metastatic gastroesophageal cancer. The additional anti-cancer therapy may include, for example, trifluridine and / or tipiracil. The additional anti-cancer therapy may include, for example, combination therapy with trifluridine and tipiracil (e.g., in the case of EGJ adenocarcinoma).

[0119] The population of modified T cells and the one or more additional anti-cancer therapies may be administered in any order and at any number of times. As described above, the population of modified T cells may be administered, for example, as a single dose. The additional anti-cancer therapy can be administered, for example, (a) before the modified T cells, (b) simultaneously with the modified T cells, and / or (c) after the modified T cells. For example, the additional anti-cancer therapy may be administered, for example, (a); (b); (c); (a) and (b); (a) and (c); (b) and (c); or (a), (b), and (c). When the additional anti-cancer therapy is administered simultaneously with the population of modified T cells, the additional anti-cancer therapy and the population of modified T cells may be included in the same composition or in separate compositions. In a preferred embodiment of the present invention, the administration of the additional anti-cancer therapy begins before or after the administration of the population of modified T cells.

[0120] Administration of an additional anti-cancer therapy simultaneously with the engineered T cells can refer to administering the additional anti-cancer therapy and the engineered T cells substantially simultaneously. In other words, the dose of the additional anti-cancer therapy can be administered approximately simultaneously with the dose of the population of engineered T cells. For example, the dose of the additional anti-cancer therapy can be administered within about 2 hours, about 4 hours, about 6 hours, about 8 hours, about 12 hours, or about 24 hours of the dose of the population of engineered T cells.

[0121] Administration of an additional anti-cancer therapy prior to the engineered T cells can refer to administration of the anti-cancer therapy at any time prior to the engineered T cells. In other words, the dose of the additional anti-cancer therapy can be administered at any time prior to the dose of the population of engineered T cells. For example, the dose of the additional anti-cancer therapy can be administered about 24 hours or more, about 36 hours or more, about 48 hours or more, about 72 hours or more, about 4 days or more, about 5 days or more, about 6 days or more, about 1 week or more, about 2 weeks or more, about 3 weeks or more, or about 4 weeks or more, about 5 weeks or more, about 6 weeks or more, about 8 weeks or more, or about 12 weeks or more prior to the population of engineered T cells. The additional anti-cancer therapy can also be administered prior to the engineered T cells, for example, to initiate treatment while generating autologous engineered T cells.

[0122] Administration of an additional anti-cancer therapy after the engineered T cells can refer to administration of the anti-cancer therapy at any time after the engineered T cells. In other words, the dose of the additional anti-cancer therapy can be administered at any time after the dose of the population of engineered T cells. For example, the dose of the additional anti-cancer therapy can be administered about 24 hours or more, about 36 hours or more, about 48 hours or more, about 72 hours or more, about 4 days or more, about 5 days or more, about 6 days or more, about 1 week or more, about 2 weeks or more, about 3 weeks or more, or about 4 weeks or more, about 5 weeks or more, about 6 weeks or more, about 8 weeks or more, or about 12 weeks or more after the population of engineered T cells. The additional anti-cancer therapy can be administered after the engineered T cells, for example, to initiate treatment while generating autologous engineered T cells.

[0123] If the additional anticancer therapy is administered before the population of modified T cells, the additional anticancer therapy may be continued after the administration of the modified T cells. Thus, in one embodiment of the present disclosure, the administration of the additional anticancer therapy is initiated before the administration of the population of modified T cells and continued after the administration of the population of modified T cells. In other words, a dose of the additional anticancer therapy may be administered before the dose of the population of modified T cells, and one or more additional doses of the additional anticancer therapy may be administered afterwards. The dose of the additional anticancer therapy may be administered, for example, according to a known treatment regimen of the additional anticancer therapy. The purpose of the additional dose of the additional anticancer therapy may be to maintain the effect achieved by the administration of the initial dose. Each of the one or more additional doses may include the same additional anticancer therapy as the initial dose, or may include an additional anticancer therapy different from the initial dose.

[0124] When an additional anti-cancer therapy is administered simultaneously with the population of modified T cells, the additional anti-cancer therapy may be continued after administration of the modified T cells. Thus, in one embodiment of the present disclosure, administration of the additional anti-cancer therapy is initiated simultaneously with administration of the population of modified T cells and continues after administration of the population of modified T cells. In other words, a dose of the additional anti-cancer therapy may be administered simultaneously with a dose of the population of modified T cells, and one or more additional doses of the additional anti-cancer therapy may be administered later. The dose of the additional anti-cancer therapy may be administered, for example, according to a known treatment regimen of the additional anti-cancer therapy. The purpose of the additional dose of the additional anti-cancer therapy may be to maintain the effect achieved by administration of the initial dose. Each of the one or more additional doses may include the same additional anti-cancer therapy as the initial dose, or may include an additional anti-cancer therapy different from the initial dose.

[0125] When an additional anticancer therapy is administered after the population of modified T cells, the additional anticancer therapy may be continued after the initial administration. Thus, in one embodiment of the present disclosure, the administration of the additional anticancer therapy begins after the administration of the population of modified T cells and continues after the initial administration of the additional anticancer therapy. In other words, a dose of the additional anticancer therapy may be administered after the dose of the population of modified T cells, and one or more additional doses of the additional anticancer therapy may be administered later. The dose of the additional anticancer therapy may be administered, for example, according to a known treatment regimen of the additional anticancer therapy. The purpose of the additional dose of the additional anticancer therapy may be to maintain the effect achieved by the administration of the initial dose. Each of the one or more additional doses may include the same additional anticancer therapy as the initial dose, or may include an additional anticancer therapy different from the initial dose.

[0126] In either case, one or more additional doses of the additional anticancer therapy may be administered at any suitable interval. Any number of additional doses of the additional anticancer therapy may be administered, such as 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 15 or more, 20 or more, 25 or more, 30 or more, 35 or more, 40 or more, 45 or more, or 50 or more additional doses. Additional doses may be administered until disease progression, unacceptable toxicity, withdrawal of consent, or death. Suitable dosing intervals for the additional anticancer therapy are known in the art and may be specific to the identity of the additional anticancer therapy.

[0127] For example, in the case of paclitaxel, one or more additional doses may be administered, for example, on days 1, 8, and 15 of a 28-day cycle. The one or more additional doses may be administered approximately once every week (starting one week after administration of the first dose). Paclitaxel may be administered in a four-week cycle, administered approximately once every three weeks followed by one week without administration (3Q4W).

[0128] In a preferred embodiment of the present disclosure, the additional anticancer therapy comprises paclitaxel and / or ramucirumab. For example, the additional anticancer therapy can comprise paclitaxel, ramucirumab, or both paclitaxel and ramucirumab. Preferably, the additional anticancer therapy comprises paclitaxel and ramucirumab. Paclitaxel and / or ramucirumab (e.g., paclitaxel and ramucirumab) may, for example, be administered before the population of engineered T cells. Paclitaxel and / or ramucirumab (e.g., paclitaxel and ramucirumab) may, for example, be administered after the population of engineered T cells. As described below, the individual may also be further administered a checkpoint inhibitor (e.g., a PD-1 axis binding antagonist such as nivolumab or pembrolizumab). The checkpoint inhibitor (e.g., nivolumab or pembrolizumab) may, for example, be administered after the population of engineered T cells. The checkpoint inhibitor (e.g., nivolumab or pembrolizumab) may be administered, for example, after paclitaxel and / or ramucirumab (e.g., paclitaxel and ramucirumab). The checkpoint inhibitor (e.g., nivolumab) may be administered, for example, after (1) the population of engineered T cells and (2) paclitaxel and / or ramucirumab (e.g., paclitaxel and ramucirumab).

[0129] Checkpoint inhibitors The method can include administering a checkpoint inhibitor to the individual. Thus, the method can include administering to the individual (i) a population of engineered T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor (TCR) capable of binding to a peptide antigen of MAGE-A4, and (ii) a checkpoint inhibitor. In other words, the method can include combination treatment of the individual with (i) a population of engineered T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor (TCR) capable of binding to a peptide antigen of MAGE-A4, and (ii) a checkpoint inhibitor. As noted above, the combination can include an additional anti-cancer therapy, such as chemotherapy.

[0130] The checkpoint inhibitor may be administered to an individual in the same selection of treatment as the population of modified T cells. As noted above, a "selection" of treatment may refer to a particular treatment regimen for a cancer that has failed a curative treatment or for which a curative treatment may be inappropriate. Thus, a checkpoint inhibitor and a population of modified T cells may be administered as part of the same treatment regimen for a cancer that has failed a curative treatment or for which a curative treatment may be inappropriate. The checkpoint inhibitor and the population of modified T cells may be administered as part of a first-line treatment regimen. The checkpoint inhibitor and the population of modified T cells may be administered as part of a second-line treatment regimen. A second-line treatment regimen may be employed, for example, after failure of a first-line treatment regimen. The checkpoint inhibitor and the population of modified T cells may be administered as part of a third-line treatment regimen. A third-line treatment regimen may be employed, for example, after failure of a second-line treatment regimen.

[0131] Checkpoint inhibitor therapy is a form of cancer immunotherapy. This therapy targets immune checkpoints, which are important regulators of the immune system that, when stimulated, can dampen the immune response to immunological stimuli. Checkpoint therapy can block inhibitory checkpoints and restore immune system function. Checkpoint inhibitors can target the molecules CTLA4, PD-1, and PD-L1.

[0132] Checkpoint inhibitors can target, for example, CTLA4. That is, checkpoint inhibitors can include CTLA4 blockers. CTLA4 blockers are known in the art and include, for example, ipilimumab.

[0133] Preferably, the checkpoint inhibitor comprises a PD-1 axis-binding antagonist, which, as mentioned above, is included in the "standard of care" for gastroesophageal cancer (e.g., nivolumab or pembrolizumab).

[0134] Programmed cell death protein 1 (PD-1, also known as CD279) is a protein expressed on the surface of T cells that regulates immune responses by maintaining T cell homeostasis. Ligation of PD-1 with one of its ligands (PD-L1 or PD-L2) mediates inhibitory signals within T cells. Specifically, PD-1-generated signals block phosphorylation of key TCR signaling intermediates, thereby terminating early TCR signaling and reducing T cell activation. This results in a reduction in T cell effector functions (e.g., proliferation, cytotoxicity, and cytokine production) and an impaired ability to transition to memory T cells.

[0135] PD-L1 and PD-L2 are members of the B7 family. PD-L1 protein is upregulated on certain activated immune cells (e.g., macrophages, dendritic cells, T cells, and B cells) and is also expressed in certain normal tissues. PD-L1 is also highly expressed in many cancers. PD-L2 is expressed primarily on dendritic cells and in some tumors. Because many cancers express PD-1 ligands, the PD-1 axis has an established role in cancer immune evasion and tumor resistance.

[0136] When PD-1 ligands are expressed by cancers, such as solid tumors, the tumor microenvironment becomes immunosuppressive. This may inhibit the function of engineered T cells that infiltrate the tumor. Endogenous anti-tumor T cell responses may also be inhibited. Thus, tumors are more likely to evade the immune system. In the present disclosure, PD-1 axis-binding antagonists can be administered to negate the suppressive effects of PD-L1 and / or PD-L2 expression in the tumor microenvironment. By negating suppression, the function of engineered and / or endogenous T cells can be sustained. That is, administration of a PD-1 axis-binding antagonist can sustain the function of the engineered T cells and / or their progeny included in the administration. Administration of a PD-1 axis-binding antagonist can sustain the function of endogenous T cells in an individual. Administration of a PD-1 axis-binding antagonist can sustain the function of the engineered T cells (and / or their progeny) included in the administration and endogenous T cells in the individual. In either case, endogenous T cells may be included in, for example, the tumor microenvironment. For example, the endogenous T cells may be tumor-infiltrating lymphocytes (TILs).

[0137] Sustained T cell function can refer, for example, to maintaining, restoring, and / or enhancing T cell function. Sustained T cell function can refer, for example, to sustaining T cell activation. In this way, the duration of an effective T cell response can be extended. In other words, sustained activation can be associated with improved duration of effector function (such as cytokine production, cytotoxicity, and / or proliferation). Sustained activation can also support the ability of T cells to transition to memory T cells. The generation of memory T cells is advantageous because it can enable anti-tumor immunity to be maintained for a long period of time, for example, for several months or years. Methods for determining activation, cytokine production, cytotoxicity, proliferation, and the generation of memory T cells are well known in the art.

[0138] Administration of PD-1 axis-binding antagonists can sustain the function of engineered T cells and / or endogenous T cells by reducing exhaustion. Exhaustion can be reduced within a population of engineered T cells and / or in T cells that are progeny of the population of engineered T cells. Exhaustion can be reduced within endogenous T cells in an individual. Exhaustion can be reduced (i) within a population of engineered T cells and / or in T cells that are progeny of the population of engineered T cells, and (ii) within endogenous T cells in an individual. Exhausted T cells typically express high levels of PD-1 and experience loss of function. For example, exhausted T cells may have a reduced ability to produce cytokines such as IL-2 or TNFα. Exhausted T cells may have a reduced proliferative capacity. Exhausted T cells may have a reduced cytotoxic capacity. Ultimately, exhausted T cells can be targeted for destruction. Thus, exhaustion leads to loss of T cell function or even the loss of T cells, which is detrimental to tumor immunity. Therefore, reducing T cell exhaustion can improve treatment outcomes.

[0139] In the context of the present disclosure, a PD-1 axis binding antagonist is a molecule that inhibits the interaction between PD-1 and a PD-1 ligand and / or the transmission of a signal resulting from the interaction between PD-1 and a PD-1 ligand. The PD-1 ligand can be PD-L1 or PD-L2. A PD-1 axis binding antagonist can, for example, reduce or block the interaction between PD-1 and a PD-1 ligand. A PD-1 axis binding antagonist can, for example, reduce or block the transmission of a signal resulting from the interaction between PD-1 and a PD-1 ligand. A PD-1 axis binding antagonist can block, inhibit, or reduce the biological activity of PD-1 and / or a PD-1 ligand.

[0140] By inhibiting the interaction between PD-1 and its ligand and / or the transmission of signals resulting from the interaction between PD-1 and its ligand, PD-1 axis-binding antagonists can sustain (e.g., maintain, restore, or enhance) the function of endogenous T cells. Similarly, PD-1 axis-binding antagonists can sustain (e.g., maintain, restore, or enhance) the function of engineered T cells administered to an individual. Sustained function can be demonstrated, for example, by the maintenance or improvement of T cell proliferation, cytokine production, target cell killing, activation, CD28 signaling, ability to infiltrate tumors, ability to recognize and bind antigens presented by dendritic cells, and / or ability to produce interferon. In this way, PD-1 axis-binding antagonists negate the immunosuppressive properties of the tumor microenvironment.

[0141] The PD-1 axis binding antagonist can be, for example, a PD-1 binding antagonist. That is, the PD-1 axis binding antagonist can inhibit (e.g., block or reduce) the binding of PD-1 to its binding partner. For example, the PD-1 axis binding antagonist can inhibit the binding of PD-1 to PD-L1, PD-L2, or both PD-L1 and PD-L2. The PD-1 binding antagonist can be, for example, an antibody that binds to PD-1, or an antigen-binding variant or fragment thereof. The PD-1 binding antagonist can be, for example, an antibody that binds to SEQ ID NO: 14, or an antigen-binding variant or fragment thereof. Antibodies that bind to PD-1 are known in the art and include, for example, nivolumab, pembrolizumab, cemiplimab, dostallimab, JTX-4014, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, retifanlimab (INCMGA00012), AMP-224, MEDI0680 (AMP-514), sasanlimab, budigalimab, ezabenlimab (BI 754091), and zimberelimab (AB122). Thus, the PD-1 axis binding antagonist may be nivolumab, pembrolizumab, cemiplimab, dostallimab, JTX-4014, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, retifanlimab (INCMGA00012), AMP-224, MEDI0680 (AMP-514), sasanlimab, budigalimab, ezabenlimab (BI 754091), or zimberelimab (AB122), or any combination thereof. The PD-1 axis binding antagonist may be, for example, nivolumab. The PD-1 axis binding antagonist may be, for example, pembrolizumab. Alternatively, the PD-1 binding antagonist may be any immunoadhesin, protein, or oligopeptide that inhibits, reduces, blocks, or interferes with signaling resulting from the interaction of PD-1 with PD-L1 and / or PD-L2.

[0142] The heavy and light chain sequences of nivolumab are set forth in SEQ ID NOs: 17 and 18, respectively. The heavy and light chain sequences of pembrolizumab are set forth in SEQ ID NOs: 19 and 20, respectively. The heavy and light chain sequences of cemiplimab are set forth in SEQ ID NOs: 21 and 22, respectively. The heavy and light chain sequences of dostallimab are set forth in SEQ ID NOs: 31 and 32, respectively. Armed with knowledge of the heavy and light chain sequences of a given antibody, one of skill in the art can identify antigen-binding variants or fragments of the antibody using methods routinely performed in the art. An antigen-binding variant or fragment of nivolumab may, for example, comprise the three CDRs contained in SEQ ID NO: 17 and the three CDRs contained in SEQ ID NO: 18. An antigen-binding variant or fragment of pembrolizumab may, for example, comprise the three CDRs contained in SEQ ID NO: 19 and the three CDRs contained in SEQ ID NO: 20. An antigen-binding variant or fragment of cemiplimab may, for example, comprise the three CDRs contained in SEQ ID NO: 21 and the three CDRs contained in SEQ ID NO: 22. An antigen-binding variant or fragment of dostarlimab may, for example, comprise the three CDRs contained in SEQ ID NO: 31 and the three CDRs contained in SEQ ID NO: 32. Methods for identifying CDRs within a heavy or light chain sequence are routine in the art.

[0143] The PD-1 axis-binding antagonist may be, for example, a PD-L1 binding antagonist. That is, the PD-1 axis-binding antagonist can inhibit (e.g., block or reduce) the binding of PD-L1 to a binding partner. For example, the PD-1 axis-binding antagonist can inhibit the binding of PD-L1 to PD-1. The PD-L1 binding antagonist may be, for example, an antibody that binds to PD-L1 or an antigen-binding variant or fragment thereof. The PD-L1 binding antagonist may be, for example, an antibody that binds to SEQ ID NO: 15 or an antigen-binding variant or fragment thereof. Antibodies that bind to PD-L1 are known in the art and include, for example, durvalumab, atezolizumab, avelumab, BMS 936559 (MDX-1105), envafolimab (KN035), and cosibelimab (CK-301). Thus, the PD-L1 axis-binding antagonist can be durvalumab, atezolizumab, avelumab, BMS 936559 (MDX-1105), embafolimab (KN035), or cosibelimab (CK-301), or any combination thereof. Alternatively, the PD-L1 axis-binding antagonist can be any immunoadhesin, protein, or oligopeptide that inhibits, reduces, blocks, or interferes with signaling resulting from the interaction of PD-L1 with PD-1.

[0144] The heavy and light chain sequences of durvalumab are set forth in SEQ ID NOs: 23 and 24, respectively. The heavy and light chain sequences of atezolizumab are set forth in SEQ ID NOs: 25 and 26, respectively. The heavy and light chain sequences of avelumab are set forth in SEQ ID NOs: 27 and 28, respectively. The heavy and light chain sequences of BMS 936559 (MDX-1105) are set forth in SEQ ID NOs: 29 and 30, respectively. Armed with knowledge of the heavy and light chain sequences of a given antibody, one of skill in the art can identify antigen-binding variants or fragments of the antibody using methods routinely performed in the art. An antigen-binding variant or fragment of durvalumab may, for example, comprise the three CDRs contained in SEQ ID NO: 23 and the three CDRs contained in SEQ ID NO: 24. An antigen-binding variant or fragment of atezolizumab may, for example, comprise the three CDRs contained in SEQ ID NO: 25 and the three CDRs contained in SEQ ID NO: 26. An antigen-binding variant or fragment of avelumab may, for example, comprise the three CDRs contained in SEQ ID NO: 27 and the three CDRs contained in SEQ ID NO: 28. An antigen-binding variant or fragment of BMS 936559 (MDX-1105) may, for example, comprise the three CDRs contained in SEQ ID NO: 29 and the three CDRs contained in SEQ ID NO: 30. Methods for identifying CDRs within a heavy or light chain sequence are routine in the art.

[0145] The PD-1 axis-binding antagonist may be, for example, a PD-L2 binding antagonist. That is, the PD-1 axis-binding antagonist can inhibit (e.g., block or reduce) the binding of PD-L2 to a binding partner. The PD-L2 binding antagonist may be, for example, an antibody that binds to PD-L2, or an antigen-binding variant or fragment thereof. The PD-L2 binding antagonist may be, for example, an antibody that binds to SEQ ID NO: 16, or an antigen-binding variant or fragment thereof. Alternatively, the PD-L2 binding antagonist may be any immunoadhesin, protein, or oligopeptide that inhibits, reduces, prevents, or interferes with signaling resulting from the interaction of PD-L2 with PD-1.

[0146] When the PD-1 axis-binding antagonist is an antibody (such as a known antibody or its antigen-binding variant), it may be, for example, a monoclonal antibody, a human or humanized antibody, a full-length antibody, a diabody, a linear antibody, or a single-chain antibody molecule. The antibody isotype can be selected from any of the five immunoglobulin classes: IgA, IgD, IgE, IgG, and IgM, which have heavy chains designated alpha, delta, epsilon, gamma, and mu (M), respectively. Antibodies of the gamma and alpha classes may be any of the subclasses: IgG1, IgG2A, IgG2B, IgG3, IgG4, IgA1, and IgA2. When the PD-1 axis-binding antagonist is an antigen-binding fragment of an antibody, it may be, for example, an Fv, Fab, Fab', Fab'-SH, F(ab')2, or scFv.

[0147] When the PD-1 axis binding antagonist is an immunoadhesin, the immunoadhesin can comprise an adhesin domain that confers binding activity for a PD-1 axis component (e.g., PD-1, PD-L1, or PD-L2) and an immunoglobulin constant domain. The immunoglobulin constant domain can be of any isotype, such as IgG1, IgG2, IgG2A, IgG2B, IgG3, IgG4 subtypes, IgA, IgA1, IgA2, IgE, IgD, or IgM. The immunoglobulin constant domain can comprise, for example, (i) the hinge, CH2, and CH3 regions or (ii) the hinge, CH1, CH2, and CH3 regions of an immunoglobulin molecule. Thus, the immunoadhesin may comprise (a) the extracellular or PD-1-binding portion of PD-L1 or PD-L2, or the extracellular or PD-L1- or PD-L2-binding portion of PD-1 fused to (b) a constant domain of an immunoglobulin sequence.

[0148] At least 1% of gastroesophageal cancer cells from an individual may express PD-L1, e.g., at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, or at least 80%. Gastroesophageal cancer from an individual may express PD-L1 with a tumor proportion score (TPS) of 1% or more (≧), e.g., ≧2%, ≧10%, or ≧50%. The proportion of cells expressing PD-L1 can be determined by any means known to one of skill in the art, such as IHC, flow cytometry, or ELISA.

[0149] At least 1% of gastroesophageal cancer cells from an individual may express PD-L2, e.g., at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, or at least 80%. Gastroesophageal cancer cells from an individual may express PD-L2 with a tumor proportion score (TPS) of 1% or more (≧), e.g., ≧2%, ≧10%, or ≧50%. The proportion of cells expressing PD-L2 can be determined by any means known to one of skill in the art, such as IHC, flow cytometry, or ELISA.

[0150] PD-1, PD-L1, and / or PD-L2 expression in gastroesophageal cancer can have an intensity of 1+ or greater (≧), for example, ≧2+ or ≧3+. Intensity scores can be assessed by IHC staining of the tumor and are scored as follows: negative = no staining or staining in ≦10% of stained cells; 1+ = incomplete staining in ≧10% of stained cells; 2+ = weak to moderate staining in ≧10% of stained cells; strong and complete staining in ≧10% of stained cells.

[0151] The population of engineered T cells and the checkpoint inhibitor may be administered in any order and at any number of times. The population of engineered T cells may be administered, for example, as a single dose. The checkpoint inhibitor can be administered, for example, (a) before the engineered T cells, (b) simultaneously with the engineered T cells, and / or (c) after the engineered T cells. For example, the checkpoint inhibitor may be administered, for example, (a); (b); (c); (a) and (b); (a) and (c); (b) and (c); or (a), (b), and (c). When the checkpoint inhibitor is administered simultaneously with the population of engineered T cells, the checkpoint inhibitor and the population of engineered T cells may be included in the same composition or in separate compositions.

[0152] Administering a checkpoint inhibitor simultaneously with the engineered T cells can refer to administering the checkpoint inhibitor and the engineered T cells substantially simultaneously. In other words, the dose of the checkpoint inhibitor may be administered at about the same time as the dose of the population of engineered T cells. For example, the dose of the checkpoint inhibitor may be administered within about 2 hours, within about 4 hours, within about 6 hours, within about 8 hours, within about 12 hours, or within about 24 hours of the dose of the population of engineered T cells.

[0153] Administering a checkpoint inhibitor prior to the engineered T cells can refer to administering the checkpoint inhibitor at any time prior to the engineered T cells. In other words, the dose of the checkpoint inhibitor can be administered at any time prior to the dose of the population of engineered T cells. For example, the dose of the checkpoint inhibitor can be administered about 24 hours or more, about 36 hours or more, about 48 hours or more, about 72 hours or more, about 4 days or more, about 5 days or more, about 6 days or more, about 1 week or more, about 2 weeks or more, about 3 weeks or more, or about 4 weeks or more, about 5 weeks or more, about 6 weeks or more, about 8 weeks or more, or about 12 weeks or more prior to the population of engineered T cells. The checkpoint inhibitor can also be administered prior to the engineered T cells, for example, to initiate treatment while generating autologous engineered T cells.

[0154] Administering a checkpoint inhibitor after the engineered T cells can refer to administering the checkpoint inhibitor at any time after the engineered T cells. In other words, the dose of the checkpoint inhibitor can be administered at any time after the dose of the population of engineered T cells. For example, the dose of the checkpoint inhibitor can be administered about 24 hours or more, about 36 hours or more, about 48 hours or more, about 72 hours or more, about 4 days or more, about 5 days or more, about 6 days or more, about 1 week or more, about 2 weeks or more, about 3 weeks or more, or about 4 weeks or more, about 5 weeks or more, about 6 weeks or more, about 8 weeks or more, or about 12 weeks or more after the population of engineered T cells.

[0155] When the checkpoint inhibitor is administered simultaneously with the population of engineered T cells, the checkpoint inhibitor may be continued after administration of the engineered T cells. Thus, in one embodiment of the present disclosure, administration of the checkpoint inhibitor is initiated simultaneously with administration of the population of engineered T cells and continues after administration of the population of engineered T cells. In other words, a dose of the checkpoint inhibitor may be administered simultaneously with a dose of the population of engineered T cells, and one or more additional doses of the checkpoint inhibitor may be administered later. The dose of the checkpoint inhibitor may be administered, for example, according to a known therapeutic regimen for the checkpoint inhibitor. The purpose of the additional dose of the checkpoint inhibitor may be to maintain the effect achieved by administration of the initial dose. Each of the one or more additional doses may include the same checkpoint inhibitor as the initial dose, or may include a different checkpoint inhibitor than the initial dose.

[0156] If the checkpoint inhibitor is administered after the population of engineered T cells, the checkpoint inhibitor may be continued after the initial administration. Thus, in one embodiment of the present disclosure, administration of the checkpoint inhibitor begins after administration of the population of engineered T cells and continues after the initial administration of the checkpoint inhibitor. In other words, a dose of the checkpoint inhibitor may be administered after the dose of the population of engineered T cells, and one or more additional doses of the checkpoint inhibitor may be administered later. The dose of the checkpoint inhibitor may be administered, for example, according to a known therapeutic regimen for the checkpoint inhibitor. The purpose of the additional dose of the checkpoint inhibitor may be to maintain the effect achieved by administration of the initial dose. Each of the one or more additional doses may contain the same checkpoint inhibitor as the initial dose, or may contain a different checkpoint inhibitor than the initial dose.

[0157] In either case, one or more additional doses of the checkpoint inhibitor may be administered at any suitable interval. Suitable dosing intervals for checkpoint inhibitors are known in the art and may be specific to the identity of the checkpoint inhibitor. For example, in the case of a PD-1 axis binding antagonist such as nivolumab or pembrolizumab, one or more additional doses may be administered, for example, about once every two weeks (Q2W) starting two weeks after administration of the first dose. One or more additional doses may be administered, for example, about once every three weeks (Q3W) starting three weeks after administration of the first dose. One or more additional doses may be administered, for example, about once every four weeks (Q4W) starting four weeks after administration of the first dose. One or more additional doses may be administered, for example, about once every five weeks (Q5W) starting five weeks after administration of the first dose. One or more additional doses may be administered, for example, about once every six weeks (Q6W) starting six weeks after administration of the first dose. In a preferred embodiment of the present disclosure, the checkpoint inhibitor is nivolumab, and the one or more additional doses are administered about once every four weeks (Q4W) starting four weeks after administration of the first dose.

[0158] Any number of additional doses of checkpoint inhibitor may be administered, such as 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 15 or more, 20 or more, 25 or more, 30 or more, 35 or more, 40 or more, 45 or more, or 50 or more additional doses. Additional doses may be administered until disease progression, unacceptable toxicity, withdrawal of consent, or death.

[0159] Combination Treatment Protocol Examples of combination treatment protocols are shown in Figures 1-5.

[0160] Figure 1 relates to the treatment of gastroesophageal cancer that has relapsed after first-line therapy, e.g., an approved or "standard of care" first-line therapy. In this case, an individual may be administered (i) a population of modified T cells, (ii) an additional anti-cancer therapy, and optionally (iii) a PD-1 axis-binding antagonist. Lymphocyte depletion may be performed prior to administration of the modified T cell population.

[0161] The additional anti-cancer treatment can be, for example, an approved or "standard of care" second-line treatment for gastroesophageal cancer. The "standard of care" second-line treatment can be, for example, paclitaxel and / or ramucirumab. For example, the "standard of care" second-line treatment can include paclitaxel and / or ramucirumab. Treatment regimens for paclitaxel and / or ramucirumab are known in the art.

[0162] The population of engineered T cells and the additional anti-cancer therapy may be administered in either order. For example, the additional anti-cancer therapy may be administered before the population of engineered T cells according to Cohort 2A (left side of Figure 1). Alternatively, the additional anti-cancer therapy may be administered after the population of engineered T cells according to Cohort 2B (right side of Figure 1). In either case, administration of the additional anti-cancer therapy may continue after administration of the engineered T cells. In other words, one or more doses of the additional anti-cancer therapy may be administered after administration of the population of engineered T cells.

[0163] When a PD-1 axis-binding antagonist is administered, it is typically administered after the population of engineered T cells and the additional anti-cancer therapy. For example, according to Cohort 2A (left side of Figure 1), an individual may be administered the additional anti-cancer therapy, then the population of engineered T cells, and then the PD-1 axis-binding antagonist. However, it is also possible to administer the population of engineered T cells to an individual, then the additional anti-cancer therapy, and then the PD-1 axis-binding antagonist. The PD-1 axis-binding antagonist may be, for example, nivolumab or pembrolizumab.

[0164] In Figure 1, administration of engineered T cells and additional anti-cancer therapy (with or without PD-1 axis binding antagonists) can be a second-line treatment for gastroesophageal cancer.

[0165] Figure 2 relates to the treatment of gastroesophageal cancer that has recurred after second line therapy, e.g., an approved or "standard of care" second line therapy. By implication, the gastroesophageal cancer is one that has recurred after first line therapy, e.g., an approved or "standard of care" first line therapy. To treat gastroesophageal cancer that has recurred after second line therapy, an individual may be administered (i) a population of modified T cells and (ii) a checkpoint inhibitor. Lymphocyte depletion may be performed prior to administration of the modified T cell population.

[0166] The checkpoint inhibitor may include, for example, a PD-1 axis-binding antagonist. The PD-1 axis-binding antagonist may be, for example, nivolumab or pembrolizumab. Treatment regimens for PD-1 axis-binding antagonists such as nivolumab and pembrolizumab are known in the art.

[0167] The checkpoint inhibitor may be administered, for example, before the population of engineered T cells, after the population of engineered T cells, or simultaneously (or substantially simultaneously) with the engineered T cells. In one embodiment of the present disclosure, the dose of the checkpoint inhibitor may be administered approximately simultaneously with the dose of the population of engineered T cells. For example, the dose of the checkpoint inhibitor may be administered within about 2 hours, about 4 hours, about 6 hours, about 8 hours, about 12 hours, or about 24 hours of the dose of the population of engineered T cells. Administration of the checkpoint inhibitor may continue, for example, after administration of the engineered T cells. In one embodiment of the present disclosure, the dose of the checkpoint inhibitor may be administered approximately simultaneously with the dose of the population of engineered T cells, and one or more additional doses of the checkpoint inhibitor may be administered later. The dose of the checkpoint inhibitor may be administered, for example, according to known therapeutic regimens for checkpoint inhibitors. Therapeutic regimens for PD-1 axis binding inhibitors (e.g., nivolumab or pembrolizumab) are known in the art.

[0168] The protocol depicted in Figure 2 may optionally include additional anti-cancer therapy, as described above.

[0169] In Figure 2, administration of modified T cells and checkpoint inhibitors can be a third-line or subsequent-line treatment for gastroesophageal cancer.

[0170] Figures 4 and 5 relate to the treatment of gastroesophageal cancer that (A) has relapsed after curative-intent therapy for locally advanced cancer or (B) is initially diagnosed as unresectable locally advanced or metastatic cancer. In this case, the individual may be administered (i) an oxaliplatin-based therapy and (ii) a population of engineered T cells. Optionally, the individual may be administered (iii) a PD-1 axis-binding antagonist and / or an additional anti-cancer therapy. Lymphocyte depletion may be performed prior to administration of the engineered T cell population.

[0171] The PD-1 axis-binding antagonist can be, for example, an approved treatment or "standard of care" for gastroesophageal cancer. The PD-1 axis-binding antagonist can be, for example, pembrolizumab or nivolumab. The additional anti-cancer therapy can be, for example, an approved treatment or "standard of care" for gastroesophageal cancer. The additional anti-cancer therapy can include, for example, fluorouracil (5FU).

[0172] The oxaliplatin-based therapy may be administered before the population of modified T cells. Administration of the modified T cells may be followed by administration of the additional anti-cancer therapy. In other words, one or more doses of the additional anti-cancer therapy may be administered after administration of the modified T cells.

[0173] If the PD-1 axis-binding antagonist and / or additional anti-cancer therapy is administered, it is typically administered after the oxaliplatin-based therapy and the population of engineered T cells. For example, the individual may be administered the oxaliplatin-based therapy, then the engineered T cells, then the PD-1 axis-binding antagonist and / or additional anti-cancer therapy.

[0174] In Figures 4 and 5, administration of engineered T cells in combination with oxaliplatin-based therapy (and optionally additional anti-cancer therapy and / or PD-1 axis binding antagonists) can be a first-line treatment for gastroesophageal cancer.

[0175] In combination therapy, administration of engineered T cells may be advantageous. In particular, administration of engineered T cells may allow for a reduction in the dose of checkpoint inhibitors or additional anti-cancer therapy. This may result in a reduction in cytokine release syndrome (CRS) and / or cytopenias, especially when the additional anti-cancer therapy is chemotherapy.

[0176] In combination therapy, the administration of a checkpoint inhibitor or additional anticancer therapy may be advantageous. In particular, the checkpoint inhibitor or additional anticancer therapy can be administered while the autologous population of modified T cells is being generated, thereby allowing treatment to be initiated as soon as possible. This may improve treatment outcomes. The administration of a checkpoint inhibitor or additional anticancer therapy and modified T cells may also have an adjuvant effect.

[0177] Administration The population of engineered T cells may be administered to an individual as soon as possible after diagnosis of recurrent gastroesophageal cancer, e.g., recurrent gastroesophageal cancer. The population can be administered to an individual, for example, less than about 150 days after diagnosis of gastroesophageal cancer, e.g., less than about 125 days, less than about 100 days, less than about 90 days, less than about 80 days, or less than about 70 days after diagnosis of gastroesophageal cancer. The population can be administered to an individual, for example, about 30 to about 150 days after diagnosis of gastroesophageal cancer, e.g., about 40 to about 125 days, about 50 to about 100 days, about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, about 65 days, or about 60 days after diagnosis of gastroesophageal cancer.

[0178] Typically, the population of engineered T cells is administered as a single dose, however, one or more additional doses (such as two or more, three or more, four or more, or five or more) may be administered depending on patient factors and the discretion of the physician.

[0179] The population can include any number of modified T cells that is therapeutically effective. The number of modified T cells for a given individual can vary depending on factors such as the cancer being treated, the severity or stage of the cancer, the age of the patient, etc. Thus, the number administered can vary at the discretion of the physician and can be specific to each subject. In one example, the population can be about 0.8 x 10 9 ~Approx. 1.0×10 9 modified T cells, e.g., about 0.8 x 10 9 ~Approx. 1.2×10 9 modified T cells, approximately 1.2 x 10 9 ~Approx. 6×10 9 modified T cells, or approximately 1.0 x 10 9 ~About 10×109 The population may comprise, for example, 1.0 x 10 modified T cells. 9 modified T cells, approximately 5.0 x 10 9 modified T cells, or approximately 10 x 10 9 The modified T cells may comprise:

[0180] Typically, the population of engineered T cells is administered intravenously. Any suitable route may be used, such as, for example, intramuscular, subcutaneous, intradermal, transdermal, or intraperitoneal routes.

[0181] The population of engineered T cells may be administered in combination with a checkpoint inhibitor and / or an anti-cancer therapy, as described above. The checkpoint inhibitor or additional anti-cancer therapy may be administered by any route appropriate for a given therapy, such as intravenously, intramuscularly, subcutaneously, intradermally, transdermally, or intraperitoneally. One or more doses of the checkpoint inhibitor or anti-cancer therapy may be administered, as described above.

[0182] Each dose of checkpoint inhibitor or anti-cancer therapy can contain any therapeutically effective amount of checkpoint inhibitor or anti-cancer therapy. The amount for a given individual can vary depending on factors such as the cancer being treated, the severity or stage of the cancer, the age of the patient, etc. Thus, the amount administered can vary according to the judgment of the physician and can be specific to each subject.

[0183] As an example, the checkpoint inhibitor may be a PD-1 axis binding antagonist such as nivolumab. The initial dose of the PD-1 axis binding antagonist and / or any further doses of the PD-1 axis binding antagonist may comprise about 200 mg to about 700 mg of nivolumab, e.g., about 200 mg to about 500 mg of nivolumab. For example, the initial dose of nivolumab and / or any further doses of nivolumab may comprise about 240 mg, about 360 mg, or about 480 mg of nivolumab. The initial dose of nivolumab and / or any further doses of nivolumab may comprise, for example, about 200 mg to about 500 mg of nivolumab, e.g., about 480 mg of nivolumab. In a preferred embodiment of the present disclosure, the initial dose of nivolumab comprises 240 mg, 360 mg, or 480 mg of nivolumab, and any further doses of the PD-1 axis binding antagonist comprise 240 mg nivolumab Q2W or 480 mg nivolumab Q4W. Nivolumab may be administered at 3 mg / kg Q2W.

[0184] In some embodiments of the present disclosure, the method includes administering lymphocyte-depleting chemotherapy to the individual prior to administering the population of modified T cells. That is, lymphocyte-depleting chemotherapy may be administered prior to step (a). Lymphocyte-depleting chemotherapy may be administered, for example, from about 14 days before step (a) to about 1 day before step (a), for example, from about 13 days before step (a) to about 2 days before step (a), from about 12 days before step (a) to about 3 days before step (a), from about 11 days before step (a) to about 4 days before step (a), from about 10 days before step (a) to about 5 days before step (a), or from about 9 days before step (a) to about 1 day before step (a). The lymphocyte-depleting chemotherapy may be administered from about 7 days before step (a) to about 6 days before step (a), from about 8 days before step (a) to about 7 days before step (a), from about 10 days before step (a) to about 1 day before step (a), from about 9 days before step (a) to about 2 days before step (a), from about 8 days before step (a) to about 3 days before step (a), from about 7 days before step (a) to about 4 days before step (a), or from about 6 days before step (a) to about 5 days before step (a). Preferably, the lymphocyte-depleting chemotherapy is administered from about 7 days before step (a) to about 4 days before step (a). The purpose of lymphocyte-depleting chemotherapy may be to deplete the individual's lymphocyte compartment to provide space for the adoptively transferred modified T cells to expand. In this way, the effect of a given dose of modified T cells can be maximized. The lymphocyte-depleting chemotherapy may include any suitable lymphotoxic agent. Lymphocytotoxic agents and suitable dosages are known in the art. Lymphocyte-depleting chemotherapy may include, for example, fludarabine and / or cyclophosphamide. Typically, lymphocyte-depleting chemotherapy is administered intravenously. Any suitable route may be used, such as, for example, intramuscular, subcutaneous, intradermal, transdermal, or intraperitoneal routes.

[0185] Pharmaceutical and medical uses The present disclosure provides a population of modified T cells comprising a xenogeneic CD8 co-receptor and a xenogeneic T cell receptor capable of binding to a peptide antigen of MAGE-A4 for use in a method of treating gastroesophageal cancer in an individual. Any of the aspects described above in relation to the method of the disclosure may also be applied to the population for use.

[0186] The present disclosure also provides the use of a population of modified T cells in the manufacture of a medicament for use in a method of treating gastroesophageal cancer in an individual, the modified T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor capable of binding to a peptide antigen of MAGE-A4. Any of the aspects described above in relation to the method of the disclosure may also be applied to the use of this population.

[0187] The present disclosure also provides the use of a population of modified T cells in a method of treating gastroesophageal cancer in an individual, wherein the modified T cells comprise a heterologous CD8 co-receptor and a heterologous T cell receptor capable of binding to a peptide antigen of MAGE-A4. Any of the aspects described above in relation to the method of the present disclosure may also be applied to the use of this population. [Example]

[0188] Introduction ADP-A2M4CD8-specific peptide-enhanced affinity receptor (SPEAR™) T cells have been genetically engineered to target the tumor antigen MAGE-A4 under appropriate human leukocyte antigen (HLA) expression. ADP-A2M4CD8 are autologous CD4 and CD8 positive T cells transduced with a self-inactivating (SIN) lentiviral vector expressing a high-affinity MAGE-A4-specific T cell receptor (TCR) and an additional CD8α co-receptor.

[0189] The affinity-optimized TCR (ADP-A2M4 TCR) contains an alpha chain variable domain contained in SEQ ID NO: 2 and a beta chain variable domain contained in SEQ ID NO: 3. When expressed in T cells, the signal peptides are cleaved from SEQ ID NOs: 2 and 3 prior to surface expression. The A2M4 TCR targets the tumor antigen MAGE-A4 and activates engineered T cells. HLA-A * When presented in the O2-GVYDGREHTV antigen complex, it recognizes the MAGE-A4230-239 (GVYDGREHTV; SEQ ID NO: 1) peptide sequence derived from MAGE-A4.

[0190] The CD8α co-receptor contained in ADP-A2M4CD8 SPEAR™ T cells is engineered to provide additional functionality to CD4+ T cells. Because CD4+ T cells have weak effector function in response to class I antigens, the CD8α co-receptor was introduced together with the TCR to increase TCR binding avidity and enhance the polyfunctional response of genetically engineered CD4+ T cells against MAGE-A4-positive tumors. Co-expression of CD8α conferred CD8+ killer T cell potential to CD4+ helper T cells while maintaining or enhancing their helper cell potential. The addition of the CD8α co-receptor directly affected TCR binding to HLA-peptide complexes in CD4+ T cells, enhancing CD4+ T cell effector function. Thus, ADP-A2M4CD8 SPEAR™ T cells were engineered to improve ADP-A2M4-expressing T cells.

[0191] This was confirmed in preclinical in vitro assays, in which ADP-A2M4CD8 showed a clear improvement in T cell activation compared with ADP-A2M4-expressing T cells (when cultured with antigen-positive cells), particularly as measured by increased CD40L surface expression in the CD4+ fraction. The inclusion of dendritic cells (DCs) in the cocultures resulted in a significant improvement in ADP-A2M4CD8 T cells. Compared to cultures containing ADP-A2M4 cells, cytokine release from both DCs (IL-12, MIG) and T cells (IFNγ, IL-2, and other Th1 cells) was improved. Furthermore, transduction with ADP-A2M4CD8 resulted in the conversion of CD4+ T cells from being unable to kill MAGE-A4-positive 3D microspheres to having effective cytotoxic function. Thus, ADP-A2M4CD8-transduced CD4+ T cells exhibit not only CD4+ helper function but also improved T cell effector function.

[0192] Although 77% of individuals with gastroesophageal cancer survive 5 years after diagnosis, treatment options are suboptimal for individuals with advanced or metastatic disease. Fewer than 25% of individuals treated with chemotherapy progress to third-line therapy, and the median progression-free survival with third-line therapy is around 4-7 months.

[0193] Improved regimens for treating gastroesophageal cancer are desirable. Results of a preliminary clinical trial of ADP-A2M4CD8 in 13 individuals with gastroesophageal cancer demonstrated clear antitumor activity with two confirmed responses and encouraging disease control rates (10 of 13 patients).

[0194] Examples 1 and 2 investigate regimens using ADP-A2M4CD8 as second-line or third-line treatments for gastroesophageal cancer, respectively.

[0195] Example 1. Second-line treatment of gastroesophageal cancer. Subjects are selected for treatment with ADP-A2M4CD8. Briefly, subjects eligible for selection must have previously been treated with first-line standard therapy for gastroesophageal cancer and either failed that first-line treatment or subsequently experienced a recurrence of cancer. For example, if the first-line standard therapy includes fluorouracil chemotherapy and / or platinum-based chemotherapy, selected patients typically undergo a follow-up PET and / or CT scan 5 to 8 weeks after completing treatment. This scan can identify persistent (relapsed) local disease, unresectable locally advanced disease, or metastatic disease. Subjects who received first-line standard therapy with fluorouracil chemotherapy and / or platinum-based chemotherapy may be particularly suitable for treatment.

[0196] Additionally, the selected subjects were HLA-A * 02:01, HLA-A * 02:03, or HLA-A * 02:06, or another HLA-A with the same protein sequence in the peptide-binding domain * Positive for the 02 allele. HLA-A* 02:05 Positive patients have two HLA-A * Since alloreactivity from ADP-A2M4 and ADP-A2M4CD8 to 02:05-positive cell lines has already been confirmed, they are excluded from this study. * HLA-A as the 02 allele * 02:07 or any A * Patients with any of the O2 null alleles are also excluded because these alleles result in reduced activity.

[0197] Selected subjects also have tumors in which ≧30% of tumor cells exhibit MAGE-A4 expression defined as ≧2+ by immunohistochemistry (IHC).

[0198] Autologous cells are collected by leukapheresis from a selected subject for processing and manufacturing into ADP-A2M4CD8. The heterologous TCR contained in the ADP-A2M4CD8 T cells comprises an alpha chain sequence contained in SEQ ID NO:2 and a beta chain sequence contained in SEQ ID NO:3. The heterologous CD8 co-receptor contained in the ADP-A2M4CD8 T cells comprises two CD8 alpha chains, each comprising the amino acid sequence of SEQ ID NO:10. The surface-expressed heterologous TCR and surface-expressed heterologous CD8 co-receptor do not contain signal sequences.

[0199] A baseline tumor assessment will be obtained before treatment. Subjects will then receive either (1) second-line "standard of care" (paclitaxel and ramucirumab) followed by ADP-A2M4CD8 or (2) ADP-A2M4CD8 followed by second-line "standard of care" (paclitaxel and ramucirumab). Lymphocyte-depleting chemotherapy with fludarabine and cyclophosphamide will be administered in anticipation of ADP-A2M4CD8 administration (days -7 to -4 before ADP-A2M4CD8). Following protocol (1) or (2), subjects will optionally receive nivolumab.

[0200] For subjects in protocol (1), ramucirumab may be administered on days 1 and 15, and paclitaxel may be administered on days 1, 8, and 15. Leukapheresis may be performed on days 26-28. A second cycle of ramucirumab and paclitaxel may be administered on days 29-56. A third cycle of ramucirumab and paclitaxel may be administered on days 57-84. Leukapheresis may begin on day 85. ADP-A2M4CD8 may be infused on day 92. If leukapheresis cannot be performed at the end of the first cycle of ramucirumab and paclitaxel, it may instead be performed at the end of the second cycle. ADP-A2M4CD8 may be infused after the fourth cycle of ramucirumab and paclitaxel, or as soon as ADP-A2M4CD8 becomes available after the third cycle of ramucirumab and paclitaxel.

[0201] 1×10 8 ~1×10 10 ADP-A2M4CD8 T cells are administered to the subject. The initial dose selected for ADP-A2M4CD8 is 1 x 10 9 transduced cells (range: 0.8 × 10 9 ~1.2×10 9 transduced cells) administered via a single intravenous infusion.

[0202] Subjects will be monitored immediately after infusion (Days 1-8). Subjects will be monitored weekly until Week 4 after infusion. Subjects will then be monitored at Weeks 6, 8, 12, 16, and 24, and at least every 3 months thereafter until disease progression. Tumor response will be assessed according to Response Evaluation Criteria in Solid Tumors (RECIST) v1.1. Additional data collected will include: - Core needle biopsies to directly assess the "immune landscape" within the tumor at baseline and throughout the study. - Serum cytokine levels at baseline and throughout the study period. - Humoral immune responses to tumor antigens using serum at baseline and throughout the study. - Serum antibodies against ADP-A2M4CD8 at baseline and throughout the study period. - Soluble markers of tumors and their microenvironment using liquid biopsies. For example, markers of circulating tumor cells (CTCs), exosomes, and cell-free DNA (cfDNA) produced by dying tumor cells can be used to monitor both the molecular signature of tumor burden (including target antigen expression) and immune responses. Analysis of such soluble markers allows for estimation of systemic tumor burden and genetic profiling, including MAGE-A4 mRNA expression and mutational profiling. Analysis of such soluble markers also allows for systemic assessment of immune responses. - Phenotype and activity of the genetically modified T cells before and after infusion. Relevant assays can be performed using blood and, if resection was performed, tumor. Assays include (i) phenotypic analysis to determine the lineage of T cells in the cell product and in the blood (and tumor, if resection was performed) after infusion, (ii) quantification of the senescence and activation state of immune subsets from PBMCs, (iii) analysis of gene expression or epigenetic profiles to reflect the phenotype and functional state of the cells, and / or (iv) direct functional assessment of the cells. - Persistence of the infused genetically engineered cells and its correlation with therapeutic efficacy. Persistence can be determined by data on the number of copies of genetically modified DNA per μg of DNA and / or the number of transduced cells or total lymphocytes per μL. Well-established methodologies include (i) quantification of ADP-A2M4CD8 cells by quantitative PCR of the transgene from DNA extracted from frozen PBMCs and (ii) quantification and phenotypic analysis of ADP-A2M4CD8 cells by flow cytometry, DNA analysis, and RNA analysis from frozen PBMCs.

[0203] Max 10×10 9Doses of ADP-A2M4CD8 were administered and shown to be well tolerated. New data suggest that second-line treatment of gastroesophageal cancer, when including ADP-A2M4CD8, may improve outcomes compared to "standard of care" second-line treatments for recurrent gastroesophageal cancer, such as ramucirumab in combination with paclitaxel (and optionally nivolumab).

[0204] Example 2. Third-line treatment of gastroesophageal cancer. Subjects are selected for treatment with ADP-A2M4CD8. Briefly, subjects to be selected must have previously been treated with first-line and second-line standard therapies for gastroesophageal cancer, and the first-line and second-line therapies must have been unsuccessful or the cancer must have subsequently recurred. For example, the cancer may be recurrent esophageal squamous cell carcinoma.

[0205] The selected subject also has HLA-A * 02:01, HLA-A * 02:03, or HLA-A * 02:06, or another HLA-A with the same protein sequence in the peptide-binding domain * Positive for the 02 allele. HLA-A * 02:05 Positive patients have two HLA-A * Since alloreactivity from ADP-A2M4 and ADP-A2M4CD8 to 02:05-positive cell lines has already been confirmed, they are excluded from this study. * HLA-A as the 02 allele * 02:07 or any A * Patients with either of the O2 null alleles will also be excluded, as these alleles result in reduced activity. Additionally, selected subjects will have tumors in which ≥30% of tumor cells demonstrate MAGE-A4 expression, defined as ≥2+ by immunohistochemistry (IHC).

[0206] Autologous cells are collected by leukapheresis from enrolled subjects for processing and manufacturing into ADP-A2M4CD8. The heterologous TCR contained in the ADP-A2M4CD8 T cells comprises an alpha chain sequence contained in SEQ ID NO:2 and a beta chain sequence contained in SEQ ID NO:3. The heterologous CD8 co-receptor contained in the ADP-A2M4CD8 T cells comprises two CD8 alpha chains, each comprising the amino acid sequence of SEQ ID NO:10. The surface-expressed heterologous TCR and surface-expressed heterologous CD8 co-receptor do not contain signal sequences.

[0207] A baseline tumor assessment was obtained before treatment. Subjects then received third-line "standard of care" ADP-A2M4CD8, typically a PD-1 axis-binding antagonist such as pembrolizumab. Lymphocyte-depleting chemotherapy with fludarabine and cyclophosphamide was administered in anticipation of ADP-A2M4CD8 administration (days -7 to -4 before ADP-A2M4CD8).

[0208] 1×10 8 ~1×10 10 ADP-A2M4CD8 T cells are administered to the subject. The initial dose selected for ADP-A2M4CD8 is 1 x 10 9 transduced cells (range: 0.8 × 10 9 ~1.2×10 9 transduced cells) administered via a single intravenous infusion.

[0209] Subjects will be monitored immediately after infusion (Days 1-8). Subjects will be monitored weekly until Week 4 after infusion. Subjects will then be monitored at Weeks 6, 8, 12, 16, and 24, and at least every 3 months thereafter until disease progression. Tumor response will be assessed according to Response Evaluation Criteria in Solid Tumors (RECIST) v1.1. Additional data collected may include: - Core needle biopsies to directly assess the "immune landscape" within the tumor at baseline and throughout the study. - Serum cytokine levels at baseline and throughout the study period. - Humoral immune responses to tumor antigens using serum at baseline and throughout the study. - Serum antibodies against ADP-A2M4CD8 at baseline and throughout the study period. - Soluble markers of tumors and their microenvironment using liquid biopsies. For example, markers of circulating tumor cells (CTCs), exosomes, and cell-free DNA (cfDNA) produced by dying tumor cells can be used to monitor both the molecular signature of tumor burden (including target antigen expression) and immune responses. Analysis of such soluble markers allows for estimation of systemic tumor burden and genetic profiling, including MAGE-A4 mRNA expression and mutational profiling. Analysis of such soluble markers also allows for systemic assessment of immune responses. - Phenotype and activity of the genetically modified T cells before and after infusion. Relevant assays can be performed using blood and, if resection was performed, tumor. Assays include (i) phenotypic analysis to determine the lineage of T cells in the cell product and in the blood (and tumor, if resection was performed) after infusion, (ii) quantification of the senescence and activation state of immune subsets from PBMCs, (iii) analysis of gene expression or epigenetic profiles to reflect the phenotype and functional state of the cells, and / or (iv) direct functional assessment of the cells. - Persistence of the infused genetically engineered cells and its correlation with therapeutic efficacy. Persistence can be determined by data on the number of copies of genetically modified DNA per μg of DNA and / or the number of transduced cells or total lymphocytes per μL. Well-established methodologies include (i) quantification of ADP-A2M4CD8 cells by quantitative PCR of the transgene from DNA extracted from frozen PBMCs and (ii) quantification and phenotypic analysis of ADP-A2M4CD8 cells by flow cytometry, DNA analysis, and RNA analysis from frozen PBMCs.

[0210] Max 10×10 9Doses of ADP-A2M4CD8 were administered and shown to be well tolerated. New data suggest that third-line treatment of gastroesophageal cancer, when including ADP-A2M4CD8, may improve treatment outcomes compared with "standard of care" third-line treatments for gastroesophageal cancer, such as PD-1 axis antagonist monotherapy.

[0211] Example 3. First-line treatment of gastroesophageal cancer. Subjects are selected for treatment with ADP-A2M4CD8. Briefly, subjects must have (A) relapsed after curative-intent treatment for locally advanced cancer, or (B) been initially diagnosed with unresectable locally advanced or metastatic cancer.

[0212] Additionally, the selected subjects were HLA-A * 02:01, HLA-A * 02:03, or HLA-A * 02:06, or another HLA-A with the same protein sequence in the peptide-binding domain * Positive for the 02 allele. HLA-A * 02:05 Positive patients have two HLA-A * Since alloreactivity from ADP-A2M4 and ADP-A2M4CD8 to 02:05-positive cell lines has already been confirmed, they are excluded from this study. * HLA-A as the 02 allele * 02:07 or any A * Patients with any of the O2 null alleles are also excluded because these alleles result in reduced activity.

[0213] Selected subjects also have tumors in which ≧30% of tumor cells exhibit MAGE-A4 expression defined as ≧2+ by immunohistochemistry (IHC).

[0214] Autologous cells are collected by leukapheresis from a selected subject for processing and manufacturing into ADP-A2M4CD8. The heterologous TCR contained in the ADP-A2M4CD8 T cells comprises an alpha chain sequence contained in SEQ ID NO:2 and a beta chain sequence contained in SEQ ID NO:3. The heterologous CD8 co-receptor contained in the ADP-A2M4CD8 T cells comprises two CD8 alpha chains, each comprising the amino acid sequence of SEQ ID NO:10. The surface-expressed heterologous TCR and surface-expressed heterologous CD8 co-receptor do not contain signal sequences.

[0215] A baseline tumor assessment is obtained prior to treatment. Subjects are then administered an oxaliplatin-based regimen followed by ADP-A2M4CD8. Subjects may also receive (i) a PD-1 axis binding antagonist, such as nivolumab or pembrolizumab, and / or (ii) fluorouracil (5FU). Lymphocyte-depleting chemotherapy with fludarabine and cyclophosphamide is administered prior to ADP-A2M4CD8 (days -7 to -4 before ADP-A2M4CD8).

[0216] 1×10 8 ~1×10 10 ADP-A2M4CD8 T cells are administered to the subject. The initial dose selected for ADP-A2M4CD8 is 1 x 10 9 transduced cells (range: 0.8 × 10 9 ~1.2×10 9 transduced cells) administered via a single intravenous infusion.

[0217] Subjects will be monitored immediately after infusion (Days 1-8). Subjects will be monitored weekly until Week 4 after infusion. Subjects will then be monitored at Weeks 6, 8, 12, 16, and 24, and at least every 3 months thereafter until disease progression. Tumor response will be assessed according to Response Evaluation Criteria in Solid Tumors (RECIST) v1.1. Additional data collected will include: - Core needle biopsies to directly assess the "immune landscape" within the tumor at baseline and throughout the study. - Serum cytokine levels at baseline and throughout the study period. - Humoral immune responses to tumor antigens using serum at baseline and throughout the study. - Serum antibodies against ADP-A2M4CD8 at baseline and throughout the study period. - Soluble markers of tumors and their microenvironment using liquid biopsies. For example, markers of circulating tumor cells (CTCs), exosomes, and cell-free DNA (cfDNA) produced by dying tumor cells can be used to monitor both the molecular signature of tumor burden (including target antigen expression) and immune responses. Analysis of such soluble markers allows for estimation of systemic tumor burden and genetic profiling, including MAGE-A4 mRNA expression and mutational profiling. Analysis of such soluble markers also allows for systemic assessment of immune responses. - Phenotype and activity of the genetically modified T cells before and after infusion. Relevant assays can be performed using blood and, if resection was performed, tumor. Assays include (i) phenotypic analysis to determine the lineage of T cells in the cell product and in the blood (and tumor, if resection was performed) after infusion, (ii) quantification of the senescence and activation state of immune subsets from PBMCs, (iii) analysis of gene expression or epigenetic profiles to reflect the phenotype and functional state of the cells, and / or (iv) direct functional assessment of the cells. - Persistence of the infused genetically engineered cells and its correlation with therapeutic efficacy. Persistence can be determined by data on the number of copies of genetically modified DNA per μg of DNA and / or the number of transduced cells or total lymphocytes per μL. Well-established methodologies include (i) quantification of ADP-A2M4CD8 cells by quantitative PCR of the transgene from DNA extracted from frozen PBMCs and (ii) quantification and phenotypic analysis of ADP-A2M4CD8 cells by flow cytometry, DNA analysis, and RNA analysis from frozen PBMCs.

[0218] Max 10×10 9Doses of ADP-A2M4CD8 were administered and shown to be well tolerated. New data suggest that when first-line treatment of gastroesophageal cancer includes administration of ADP-A2M4CD8, treatment outcomes may be improved compared to "standard of care" first-line treatments such as oxaliplatin-based regimens.

[0219] Example 4. Efficacy data from the Phase 1 SURPASS trial of the next-generation T cell receptor T cell therapy ADP-A2M4CD8 in patients with advanced esophageal, gastroesophageal junction, or gastric cancer.

[0220] Autologous T cells transduced with an autoinactivating lentiviral vector expressing a MAGE-A4-specific TCR and CD8α coreceptor were obtained by leukapheresis and reinfused into patients as ADP-A2M4CD8 after leukapheresis chemotherapy.

[0221] Fifteen patients with MAGE-A4-positive esophageal cancer (n=3), gastroesophageal junction (EGJ; n=10), and gastric cancer (n=2) were treated with ADP-A2M4CD8. Three of these patients (EGJ n=2, gastric n=1) received the treatment in combination with nivolumab. Prior treatment included chemotherapy (n=15), PD-(L)1 inhibitors (n=9, 60%), anti-VEGFR agents (n=9, 60%), and anti-HER2 agents (n=4, 27%). All but one patient had liver, peritoneal, and / or retroperitoneal metastases. Patient baseline characteristics are shown in the table below:

[0222] [Table 1]

[0223] The overall response rate (ORR) according to investigator-reviewed Response Evaluation Criteria in Solid Tumors (RECIST) v1.1 was 20.0% (3 partial responses; Figure 6 ).

[0224] The disease control rate was 80.0% (partial response in 3 cases and stable disease in 9 cases).

[0225] Response duration ranged from 5.0 to 29.3 weeks. Data shown in Figure 6 represent the change from baseline SLD to disease progression or prior to surgical resection.

[0226] Sequence Listing SEQ ID NO:1-MAGE-A4230-239 GVYDGREHTV SEQ ID NO:2 - MAGE-A4 TCR alpha chain (CDRs are bold and underlined VSPFSN, LTFSEN, CVVSGGTDSWGKLQF, signal sequence is italic and underlined MKKHLTTFLVILWLYFYRGNG) MKKHLTTFLVILWLYFYRGNG KNQVEQSPQSLIILEGKNCTLQCNYT VSPFSN LRWYKQDTGRGPVSLTI LTFSEN TKSNGRYTATLDADTKQSSLHITASQLSDSASYI CVVSGGTDSWGKLQF GAGTQVVVTPDIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSSGSRAKR SEQ ID NO: 3 - MAGE-A4 TCR beta chain (CDRs are bold underlined KGHDR, SFDVKD, CATSGQGAYEEQFF, signal sequence is italic underlined MASLLFFCGAFYLLGTGSMDA) MASLLFFCGAFYLLGTGSMDA DVTQTPRNRITKTGKRIMLECSQT KGHDR MYWYRQDPGLGLRLIYY SFDVKD INKGEISDGYSVSRQAQAKFSLSLESAIPNQTALYF CATSGQGAYEEQFFGPGTRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG SEQ ID NO:4 - MAGE-A4 TCR alpha chain CDR1 VSPFSN SEQ ID NO:5 - MAGE-A4 TCR alpha chain CDR2 LTFSEN SEQ ID NO:6 - MAGE-A4 TCR alpha chain CDR3 CVVSGGTDSWGKLQF SEQ ID NO:7-MAGE-A4 TCR beta chain CDR1 KGHDR SEQ ID NO:8 - MAGE-A4 TCR beta chain CDR2 SFDVKD SEQ ID NO: 9 - MAGE-A4 TCR beta chain CDR3 CATSGQGAYEEQFF SEQ ID NO: 10 - CD8 alpha chain (CDR-like loops are bold and underlined VLLSNPTSG, YLSQNKPK, LSNSIM, signal sequence is italic and underlined ALPVTALLLPLALLLHAARP) ALPVTALLLPLALLLHAARP SQFRVSPLDRTWNLGETVELKCQ VLLSNPTSG CSWLFQPRGAAASPTFLL YLSQNKPK AAEGLDTQRFSGKRLGDTFVLTLSDFRRENEGYYFCSA LSNSIM YFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRNRRRVCKCPRPVVKSGDKPSLSARYV SEQ ID NO:11 - CD8 alpha chain CDR1 VLLSNPTSG SEQ ID NO: 12 - CD8 alpha chain CDR2 YLSQNKPK SEQ ID NO: 13 - CD8 alpha chain CDR3 LSNSIM SEQ ID NO: 14 - PD1 - human programmed cell death protein (human) MQIPQAPWPVVWAVLQLGWRPGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCFSNTS ESFVLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGRDFHMSVVARRNDSGT YLCGAISLAPKAQIKESLRAELRVTERRAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGS LVLLVWVLAVICSRAARGTIGARRTGQPLKEDPSAVPVFSVDYGELDFQWREKTPEPPVP CVPEQTEYATIVFPSGMGTSSPARRGSADGPRSAQPLRPEDGHCSWPL SEQ ID NO: 15 - PD1L1 - human programmed cell death 1 ligand 1 (human) MRIFAVFIFMTYWHLLNAFTVTVPKDLYVVEYGSNMTIECKFPVEKQLDLAALIVYWEME DKNIIQFVHGEEDLKVQHSSYRQRARLLKDQLSLGNAALQITDVKLQDAGVYRCMISYGG ADYKRITVKVNAPYNKINQRILVVDPVTSEHELTCQAEGYPKAEVIWTSSDHQVLSGKTT TTNSKREEKLFNVTSTLRINTTTNEIFYCTFRRLDPEENHTAELVIPELPLAHPPNERTH LVILGAILLCLGVALTFIFRLRKGRMMDVKKCGIQDTNSKKQSDTHLEET SEQ ID NO: 16 - PD1L2 - human programmed cell death 1 ligand 2 (human) MIFLLLMLSLELQLHQIAALFTVTVPKELYIIEHGSNVTLECNFDTGSHVNLGAITASLQ KVENDTSPHRERATLLEEQLPLGKASFHIPQVQVRDEGQYQCIIIYGVAWDYKYLTLKVK ASYRKINTHILKVPETDEVELTCQATGYPLAEVSWPNVSVPANTSHSRTPEGLYQVTSVL RLKPPPGRNFSCVFWNTHVRELTLASIDLQSQMEPRTHPTWLLHIFIPFCIIAFIFIATV IALRKQLCQKLYSSKDTTKRPVTTTKREVNSAI SEQ ID NO: 17 - Nivolumab heavy chain sequence QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRYY ADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVTVSSASTKGPS VFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKP KDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPSQEEMTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSV MHEALHNHYTQKSLSLSLGK SEQ ID NO: 18 - Nivolumab light chain sequence EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPA RFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQGTKVEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 19 - Pembrolizumab heavy chain sequence QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQGLEWMGGINPSNGGTNF NEKFKNRVTLTTTDSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMGFDYWGQGTTVTVSS ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSV FLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPSQEEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEG NVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO:20 - Pembrolizumab light chain sequence EIVLTQSPATLSLSPGERATLSCRASKGVSTSGYSYLHWYQQKPGQAPRLLIYLASYLES GVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQHSRDLPLTFGGGGTKVEIKRTVAAPSVF IFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 21 - Cemiplimab heavy chain sequence EVQLLESGGVLVQPGGSLRLSCAASGFTFSNFGMTWVRQAPGKGLEWVSGISGGGRDTYF ADSVKGRFTISRDNSKNTLYLQMNSLKGEDTAVYYCVKWGNIYFDYWGQGTLVTVSSAST KGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY SLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLF PPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPSQEEMTKNQV SLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVF SCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 22 - Cemiplimab light chain sequence DIQMTQSPSSLSASVGDSITITCRASLSINTFLNWYQQKPGKAPNLLIYAASSLHGGVPS RFSGSGSGTDFTLTIRTLQPEDFATYYCQQSSNTPFTFFGPGTVVDFRRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 23 - Durvalumab heavy chain sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSRYWMSWVRQAPGKGLEWVANIKQDGSEKYY VDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREGGWFGELAFDYWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPPAFEG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPSRE EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO:24 - Durvalumab light chain sequence EIVLTQSPGTLSLSPGERATLSCRASQRVSSSYLAWYQQKPGQAPRLLIYDASSRATGIP DRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSLPWTFGQGTKVEIKRTVAAPSVFIFP PSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 25 - Atezolizumab heavy chain sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYY ADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSAS TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPS VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYAST YRVVSVLTVLHQDWLNGKEYKCKVSNKALAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ GNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 26 - Atezolizumab light chain sequence DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPS RFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 27 - Avelumab heavy chain sequence EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYIMMWVRQAPGKGLEWVSSIYPSGGITFY ADTVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARIKLGTVTTVDYWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGG PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALAPIEKTISKAKGQPREPQVYTLPPSRDE LTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 28 - Avelumab light chain sequence QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYDVSNRPSGV SNRFSGSKSGNTASLTISGLQAEDEADYYCSSYTSSSTRVFGTGTKVTVLGQPKANPTVT LFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASS YLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQ ID NO: 29 - MDX1105 heavy chain sequence QVQLVQSGAEVKKPGSSVKVSCKTSGDTFSTYAISWVRQAPGQGLEWMGGIIPIFGKAHYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYFCARKFHFVSGSPFGMDVWG QGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT SEQ ID NO: 30 - MDX1105 light chain sequence EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPTFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO:31 - Dostallimab heavy chain sequence EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVSTISGGGSYTYYQDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASPYYAMDYWGQGT TVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPP CPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 32 - Dostallimab light chain sequence DIQLTQSPSFLSAYVGDRVTITCKASQDVGTAVAWYQQKPGKAPKLLIYWASTLHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQHYSSYPWTFGQGTKLEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

Claims

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

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

3. (a) administration of the additional anti-cancer therapy is initiated prior to administration of the population of modified T cells and, optionally, is continued after administration of the population of modified T cells; or 3. The method of claim 2, wherein (b) administration of the additional anti-cancer therapy is initiated after administration of the modified T cell population.

4. 4. The method of claim 2 or 3, wherein the anti-cancer therapy comprises chemotherapy and / or targeted therapy.

5. 5. The method of any one of claims 2 to 4, wherein the anti-cancer therapy comprises (i) paclitaxel and / or ramucirumab or (ii) oxaliplatin.

6. administering to said individual a checkpoint inhibitor, and optionally (a) the population of modified T cells and the checkpoint inhibitor are administered in the same treatment; and / or (b) The method of any one of the preceding claims, wherein the checkpoint inhibitor comprises a programmed death-1 (PD-1) axis binding antagonist, optionally nivolumab or pembrolizumab.

7. (a) administration of the checkpoint inhibitor is initiated concurrently with and continued after administration of the population of modified T cells; and / or 7. The method of claim 6, further comprising (b) administering to the individual an additional anti-cancer therapy, wherein administration of the checkpoint inhibitor is initiated after administration of the additional anti-cancer therapy and the population of modified T cells.

8. 10. The method of any one of the preceding claims, wherein the gastroesophageal cancer has recurred after treatment with curative intent for locally advanced cancer or is a first diagnosis of unresectable locally advanced or metastatic cancer.

9. (a) administering oxaliplatin to the individual, wherein administration of oxaliplatin is initiated prior to administration of the population of modified T cells and, optionally, continued after administration of the population of modified T cells; and 10. The method of claim 8, comprising (b) administering to the individual a checkpoint inhibitor and / or an additional anti-cancer therapy, wherein administration of the checkpoint inhibitor and / or additional anti-cancer therapy begins after administration of the population of modified T cells, and optionally wherein the checkpoint inhibitor is a PD-1 axis binding antagonist and / or the additional anti-cancer therapy is chemotherapy, such as fluorouracil.

10. The method of any one of claims 1 to 7, wherein the gastroesophageal cancer is recurrent gastroesophageal cancer.

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

12. (a) administering to the individual an additional anti-cancer therapy, wherein administration of the additional anti-cancer therapy is initiated prior to and continued after administration of the population of modified T cells, and optionally further comprising administering to the individual a checkpoint inhibitor, wherein administration of the checkpoint inhibitor is initiated after administration of the additional anti-cancer therapy and the population of modified T cells; or (b) administering to the individual an additional anti-cancer therapy, wherein administration of the additional anti-cancer therapy begins after administration of the population of modified T cells, and optionally further administering to the individual a checkpoint inhibitor, wherein administration of the checkpoint inhibitor begins after administration of the additional anti-cancer therapy and the population of modified T cells; 12. The method of claim 10 or 11, optionally wherein (i) the anticancer therapy comprises paclitaxel and ramucirumab, and / or (ii) the checkpoint inhibitor comprises nivolumab or pembrolizumab.

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

14. administering to said individual a checkpoint inhibitor; (a) administration of the checkpoint inhibitor is initiated concurrently with and continued after administration of the population of modified T cells; or (b) administration of the checkpoint inhibitor is initiated after administration of the population of modified T cells; 14. The method of claim 13, optionally wherein the checkpoint inhibitor comprises nivolumab or pembrolizumab.

15. (a) the xenogeneic TCR binds to GVYDGREHTV (SEQ ID NO: 1) complexed with an HLA molecule; (b) the heterologous TCR comprises an alpha chain amino acid sequence having at least 80% sequence identity to SEQ ID NO:2 and a beta chain amino acid sequence having at least 80% sequence identity to SEQ ID NO:3; and / or (c) The method of any one of the preceding claims, wherein the CD8 co-receptor is CD8α.

16. the modified T cells are autologous with respect to the individual, and optionally, the method comprises: (a) obtaining peripheral blood mononuclear cells (PBMCs) from said individual; (b) selecting T cells from the PBMCs; (c) modifying the selected T cells to express the heterologous CD8 co-receptor and the heterologous TCR. generating the population by 10. The method of any one of the preceding claims, further optionally, wherein the gastroesophageal cancer is recurrent gastroesophageal cancer and one or more of steps (a) to (c) are performed before recurrence.

17. 10. A population of modified T cells comprising a xenogeneic CD8 co-receptor and a xenogeneic T cell receptor capable of binding to a peptide antigen of MAGE-A4, for use in a method according to any one of the preceding claims.

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