METHOD OF TREATMENT OF CANCER OR TUMOUR

Abstract

The present invention relates to a method of treating, preventing or delaying the progression of cancer and/or tumour in a subject comprising administering to the subject a treatment regimen comprising an effective amount of modified immunoresponsive cells expressing or presenting a heterologous T-cell receptor (TCR) having the property of binding to MAGE A4

Claims

1. A method of treating, preventing or delaying the progression of cancer and/or tumour in a subject comprising administering to the subject a treatment regimen comprising an effective amount of modified immunoresponsive cells expressing or presenting a heterologous T-cell receptor (TCR) which binds a peptide antigen of MAGE A4 comprising GVYDGREHTV, SEQ ID NO: 2, wherein the cancer and/or tumour is head and neck cancer and/or tumour or lung cancer and/or tumour.

2. (canceled)

3. The method according to claim 1, wherein (a) the heterologous TCR binds specifically and/or selectively to the peptide antigen; (b) the peptide antigen is associated with head and neck cancer and/or tumour or lung cancer and/or tumour and/or is presented by tumour and/or cancer cell or tissue; (c) the cancer and/or tumour is a MAGE A4 expressing cancer and/or tumour, and/or expresses MAGE A4 or peptide antigen thereof or a peptide antigen of MAGE A4 comprising GVYDGREHTV, SEQ ID NO: 2; and/or (d) the peptide antigen is complexed with a peptide presenting molecule, optionally major histocompatibility complex (MHC) or human leukocyte antigen (HLA), optionally class I or class II, optionally wherein the peptide presenting molecule is HLA-A*02, optionally selected from HLA*02, HLA-A*02:01, HLA-A*02:02, HLA-A*02:03, HLA-A*02:04, HLA- A*02:06, HLA-A*02:642 or HLA-A*02:07, preferably HLA-A*02:01 or HLA-A*02.

4-7. (canceled)

8. The method of claim 1, wherein the heterologous TCR binds specifically and/or selectively to the peptide antigen and/or the peptide presenting molecule and/or complex thereof.

9. The method according to claim 1 wherein the peptide antigen is presented independently of a peptide presenting molecule.

10. The method according to claim 1, wherein the heterologous TCR comprises a TCR alpha chain variable domain and a TCR beta chain variable domain, wherein: (i) the alpha chain variable domain comprises CDRs having the sequences VSPFSN (αCDR1), SEQ ID NO:11 or amino acids 48-53 of SEQ ID NO:5, or sequence having at least 50 % sequence identity thereto, LTFSEN (αCDR2), SEQ ID NO:12 or amino acids 71-76 of SEQ ID NO:5, or sequence having at least 50 % sequence identity thereto, and CVVSGGTDSWGKLQF (αCDR3), SEQ ID NO:13 or amino acids 111-125 of SEQ ID NO:5, or sequence having at least 50 % sequence identity thereto, and (ii) the beta chain variable domain comprises CDRs having the sequences KGHDR (βCDR1), SEQ ID NO:14 or amino acids 46 - 50 of SEQ ID NO:7, or sequence having at least 50 % sequence identity thereto, SFDVKD (βCDR2), SEQ ID NO:15 or amino acids 68-73 of SEQ ID NO:7, or sequence having at least 50 % sequence identity thereto, and CATSGQGAYEEQFF (βCDR3), SEQ ID NO:16 or amino acids 110 - 123 of SEQ ID NO:7 or sequence having at least 50 % sequence identity thereto.

11. The method according to claim 1, wherein the heterologous TCR comprises a TCR in which (a) the alpha chain variable domain comprises an amino acid sequence that has at least 80%, identity to SEQ ID NO:9, and/or the beta chain variable domain comprising an amino acid sequence that has at least 80% identity to SEQ ID NO:10, (b) the alpha chain variable domain comprises an amino acid sequence comprising SEQ ID NO:9, and/or the beta chain variable domain comprises SEQ ID NO:10, (c) the alpha chain comprises an amino acid sequence that has at least 80%, identity to SEQ ID NO:5, and/or the beta chain comprising an amino acid sequence that has at least 80% identity to SEQ ID NO:6, or (d) the alpha chain comprises an amino acid sequence comprising SEQ ID NO:5, and/or the beta chain comprises an amino acid sequence comprising SEQ ID NO:6.

12. The method according to claim 1, wherein the modified immunoresponsive cells expressing or presenting a heterologous TCR further express or present a heterologous co-receptor, optionally wherein the heterologous co-receptor is a CD8 co-receptor, optionally wherein the heterologous CD8 co-receptor is heterodimer or homodimer, a CD8αβ heterodimer or a CD8αα homodimer, and/or the heterologous CD8 co-receptor comprises any one of; (a) a CDR 1 of at least 80% sequence identity to amino acid sequence VLLSNPTSG, SEQ ID NO: 17, CDR 2 of at least 80% sequence identity to amino acid sequence YLSQNKPK SEQ ID NO: 18 and CDR 3 of at least 80% sequence identity amino acid sequence LSNSIM SEQ ID NO:19, (b) a CDR 1 of amino acid sequence VLLSNPTSG, SEQ ID NO:17, CDR 2 of amino acid sequence YLSQNKPK SEQ ID NO:18 and CDR 3 of amino acid sequence LSNSIM SEQ ID NO:19, (c) an amino acid sequence having at least 80% sequence identity to amino acids number 22 to 235 of SEQ ID NO: 3, or 22 to 135 of SEQ ID NO: 3, or (d) an amino acid sequence having 100% sequence identity to amino acids number 22 to 235 of sequence of SEQ ID NO: 3, or 22 to 135 of SEQ ID NO: 3.

13-14. (canceled)

15. The method according to claim 1, wherein the modified immunoresponsive cells expressing or presenting a heterologous TCR further express or present a heterologous co-stimulatory ligand, optionally 4-1BBL or CD80.

16. The method of claim 1 wherein the modified immunoresponsive cells are (a) B cells, T cells or natural killer (NK) cells, (b) T cells, optionally CD4+ T cells and/or CD8+ T cells; or (c) a population of CD4+ T cells; or CD8+ T cells, or a mixed population of CD4+ T cells and CD8+ T cells.

17. (canceled)

18. The method of claim 1, wherein the modified immunoresponsive cells are administered continuously or intermittently.

19. The method of claim 1, wherein the modified immunoresponsive cells are administered as multiple doses or as a single dose, optionally wherein the single or multiple doses are administered in one or more dosing cycles, optionally wherein the dose may be a fixed dose or a variable dose.

20. (canceled)

21. The method according to claim 1 wherein the modified immunoresponsive cells are administered at a dose of between about 500 million to about 1 billion cells, about 2 billion to about 5 billion cells or about 6 billion to about 10 billion cells.

22. The method according to claim 1 wherein the modified immunoresponsive cells are administered as; (a) a single dose in each of one or more dosing cycles, (b) one or more doses in each of one or more dosing cycles, (c) a single dose on the first day of each of one or more dosing cycles, (d) one or more doses in each of one or more dosing cycles, at least one dose being on the first day of each cycle, (e) one or more doses in each of one or more dosing cycles, at least one dose being on the first day of each cycle, (f) a single dose.

23-28. (canceled)

29. The method according to claim 1 wherein, the subject is intolerant to a standard of care treatment, optionally systemic platinum-based chemotherapy treatment.

30. The method according claim 1 wherein the head and neck or lung cancer and/or tumour has been previously unsuccessfully treated with a standard of care treatment, optionally systemic platinum-based chemotherapy treatment, or previously unsuccessfully treated with any of surgery (resection), radiation therapy, targeted therapy, immunotherapy or chemotherapy or concomitant chemotherapy with surgery (resection), radiation therapy, radiation therapy targeted therapy, checkpoint inhibitor or immunotherapy.

31. (canceled)

32. The method according to claim 1 wherein the subject has or wherein the head and neck or lung cancer and/or tumour is; primary cancer, secondary cancer, relapsed cancer or refractory cancer or recurrent cancer or locally recurrent cancer or metastatic cancer, non-resectable cancer or locally confined, cancer with no surgical or radiotherapy option or inoperable cancer optionally wherein the cancer is not amenable to transplant or loco-regional therapy, and/or wherein the head and neck cancer and/or tumour is squamous cell head and neck cancer or head and neck squamous cell carcinoma (HNSCC), optionally metastatic and/or advanced and/or locally advanced and/or recurrent squamous cell head and neck cancer or head and neck squamous cell carcinoma (HNSCC), or wherein the lung cancer and/or tumour is any one of NSCLC, squamous cell NSCLC, adenosquamous NSCLC or large cell carcinoma, optionally metastatic and/or advanced and/or locally advanced and/or recurrent NSCLC, squamous cell NSCLC, adenosquamous NSCLC or large cell carcinoma.

33-34. (canceled)

35. The method according to claim 1 wherein prior to administration of the modified immunoresponsive cells expressing or presenting a heterologous T-cell receptor (TCR) the subject undergoes lymphodepleting chemotherapy, optionally wherein the lymphodepleting chemotherapy comprises administration of cyclophosphamide and fludarabine optionally at a dose of 500 mg/m2/d x 3d cyclophosphamide and 20 mg/ m2/d x 3d fludarabine or at a dose of 600 mg/m2/d x 3d cyclophosphamide and 30 mg/ m2/d x 4d, and/or the lymphodepleting chemotherapy is administered 7 to 5 or 7 to 4 days prior to administration of the modified immunoresponsive cells expressing or presenting a heterologous T-cell receptor (TCR).

36-37. (canceled)

38. The method according to claim 1 wherein the subject has not received prior treatment for cancer and/or tumour.

39. The method according to claim 1, wherein the subject has received prior cancer and/or tumour treatment and/or has failed to respond to prior cancer and/or tumour treatment, optionally wherein the prior treatment comprises systemic and/or local therapy, optionally any one or more of surgery, radiation therapy cryotherapy, laser therapy, topical therapy and/or systemic therapy, for example any one or more of chemotherapy, hormonal therapy, targeted drugs, targeted chemotherapy, or immunotherapy, wherein the prior treatment optionally comprises: (a) a PD-L1 binding antagonist or PD-1 binding antagonist, optionally wherein the PD-1 axis binding antagonist or PD-L1 binding antagonist is an antibody; (b) an Epidermal Growth Factor Receptor Antagonist, optionally any of Cetuximab, erlotinib, gefitinib or afatinib; (c) chemotherapy comprising a platinum compound, optionally selected from any of Lipoplatin, Cisplatin, Carboplatin, Oxaliplatin, Nedaplatin, Triplatin tetranitrate, Phenanthriplatin, Satraplatin, Picoplatin; (d) chemotherapy comprising a chemotherapeutic agent selected from any of, methotrexate, capecitabine, taxane, anthracycline, paclitaxel, docetaxel, paclitaxel protein bound particles, doxorubicine, epirubicine, 5-fluorouracil, cyclophosphamide, afatinib, vincristine, etoposide or combinations thereof; or (e) chemotherapy comprising a chemotherapeutic agent selected from any of, FEC: 5-fluorouracil, epirubicine, cyclophosphamide; FAC: 5-fluorouracil, doxorubicine, cyclophosphamide; AC: doxorubicine, cyclophosphamide; EC: epirubicine, cyclophosphamide.

40-47. (canceled)

48. The method according to claim 1 wherein the treatment effectively extends or improves: (a) progression free survival, (b) time to progression, (c) duration of response, (d) overall survival, (e) objective response or objective response rate, (f) overall response or overall response rate, (g) partial response or partial response rate, (h) complete response or complete response rate; (i) stable disease rate or median stable disease (j) median progression free survival, (k) median time to progression, (1) median duration of response, (m) median overall survival; (n) median objective response or median objective response rate, (o) median overall response or median overall response rate, (p) median partial response or median partial response rate, (q) median complete response or median complete response, or (r) median stable disease rate or median stable disease, in comparison to a placebo treatment or in comparison to prior to treatment or in comparison to without treatment or in comparison to treatment comprising a standard of care, optionally systemic platinum-based chemotherapy treatment.

Description

FIGURES

[0344] FIG. 1. Table of best overall response: RECIST v1.1, responses in, head & neck cancer, and lung cancer following MAGE-A4.sup.c1032 T-cell infusion.

[0345] FIG. 2. CT data demonstrating a 50% decrease in target lesions in lung cancer at a 12 week period following MAGE-A4.sup.c1032 T-cell infusion for the cohort subject receiving a 10 billion cell infusion, upper panels showing tumour reduction, lower panels showing fluid reduction in in pleural space.

[0346] FIG. 3. Data table showing the % change reduction from baseline lesion SLC i.e. percent changes in sum of diameters in target lesions measurement (Sum of Diameters = Sum of the long diameters for non-nodal lesions and short axis for nodal lesions), responses evaluated by RECIST v1.1.

[0347] FIG. 4. Figure showing PFS and OS for treatment of sarcoma (synovial sarcoma) with MAGE-A4.sup.c1032 T-cell infusion.

[0348] FIG. 5. Table of best overall response: RECIST v1.1, responses in sarcoma (synovial sarcoma) following MAGE-A4.sup.c1032 T-cell infusion.

[0349] FIG. 6. Comparative efficacy observed in synovial sarcoma treatment; tables of % response rates: RECIST v1.1 percentage change from baseline SLD), responses in sarcoma (synovial sarcoma) following (a) standard of care treatments (b) MAGE-A4.sup.c1032 T-cell infusion.

[0350] FIG. 7. CT scans showing PR (partial response) at 24 weeks post transduced T cell infusion: CT data (lesion circled) from subject with head and neck cancer with MAGE-A4 expression (biopsy showed invasive SCC and left artenoid SCC.), previous treatment regimens had included: cisplatin, carboplatin + doxetaxel, pembrolizumab. Baseline scan showed SLD 47 mm (LN (lymph node) in chest target lesions & NT lesion in neck). Subject received 3.83 × 10.sup.9 transduced cells by infusion (ADP-A2M4 SPEAR T-cells). Recorded 36% reduction in SLD of lesion from baseline measurement at 24 weeks.

[0351] FIG. 8. CT scans showing PR (partial response) for subject with lung cancer at 20 weeks post transduced T cell infusion with tabulated data for the same subject: CT data (lesion with measurement scale) from subject with stage IV squamous cell NSLCL (PD-L1, ROS1, ). Biopsy showed MAGE-A4 antigen expression [IHC Levels: +1(0%); +2(5%); +3(95%)]. Prior cancer therapy included: surgery, systemic therapy (including chemotherapy, targeted therapy and immunotherapy); carboplatin/paclitaxel/denosumab; nivolumab; nivolumab and ipilimumab; docetaxel and ramucirumab; radiation therapy. Subject was infused with 6.5 x 10.sup.9 transduced T cells (ADP-A2M4 SPEAR T-cells). Recorded 41.7% reduction in SLD of lesion from baseline measurement at 20 weeks.

EXAMPLES

Example 1 - A Phase I Open Label, Clinical Trial Evaluating the Safety and Anti-Tumour Activity of Autologous T Cells Expressing Enhanced TCRs Specific for MAGE-A4, MAGE-A4.SUP.c1032.T in HLA-A2+ Subjects With MAGE-A4 Positive Head and Neck and Lung Tumors

Methods

[0352] The following presents an in human study of genetically engineered MAGE-A4.sup.c1032T cells in subjects with HLA-A*02 and MAGE-A4 positive inoperable locally advanced or metastatic tumors for head and neck or lung cancer.

[0353] Disease was histologically or cytogenetically confirmed and/or measurable disease recorded according to RECIST v1.1 criteria. Subjects who were eligible based on HLA type and who met MAGE-A4 criteria were screened for general health, performance status and disease stage. Following Screening, subjects meeting all eligibility criteria underwent leukapheresis to obtain cells for the manufacture of autologous MAGE-A4 TCR bearing T-cells. Eligible subjects had an ECOG Performance Status 0-1, adequate organ function and measurable disease required prior to lymphodepletion and: [0354] (a) Inoperable or metastatic (advanced) squamous cell head and neck cancer. Has received a platinum containing chemotherapy for treatment of primary tumor in adjuvant, locally advanced, or metastatic settings, or is intolerant, or refused such treatment. May have received prior immunotherapy. There is no limit on lines of prior anti-cancer therapies. [0355] (b) Histologically or cytologically confirmed diagnosis of advanced NSCLC (stage IIIB or IV) or recurrent disease. Has squamous cell, adenosquamous or large cell carcinoma. Has received at least one prior systemic therapy. Subjects whose tumors are known to have EGFR mutations or ALK gene rearrangements have failed (progressive disease or unacceptable toxicity) prior EGFR inhibitor or ALK tyrosine kinase inhibitor, respectively. Subjects with ROS-1 positive tumors have failed an ALK inhibitor (crizotinib). May have received PD-1 inhibitors. There is no limit on lines of prior anti-cancer therapies.

[0356] Exclusion of subjects is based primarily on HLA-A genotype ie if : Subject is HLA-A* 02:05 positive. Subject has HLA-A*02:07 as the sole HLA-A*02 allele (eg. A subject with HLA alleles A*02:04 and A*02:07 is eligible). Subject has any A*02 null allele (designated with an “N”, eg, A*-2:32N) as the sole HLA-A*02 allele. Excluded subjects include those with symptomatic CNS metastases.

[0357] Following leukapheresis the cells are subsequently transduced with the MAGE-A4.sup.c1032T cells (SEQ ID NO: 5, 7) specific for MAGE-A4 antigen (particularly the specific MAGE-A4 antigenic peptide SEQ ID NO:2) and the cells expanded and cryopreserved for later use. Once the MAGE-A4.sup.c1032T cells were available, subjects underwent lymphodepleting chemotherapy with cyclophosphamide plus fludarabine on Days -7 to -5, or Days -7 to -4 followed by infusion of transduced cells on Day 1.

[0358] Three subject cohorts were treated dosing with between 100 million to 5 billion transduced cells respectively with no dose escalation:

[0359] 100mn cell dose, (cyclophosphamide: 500 mg/m.sup.2/d) × 3d; (fludarabine: 20 mg/m.sup.2/d) × 3d 1bn cell dose, (cyclophosphamide: 500 mg/m.sup.2/d) × 3d; (fludarabine: 20 mg/m.sup.2/d) × 3d 5bn cell dose, (cyclophosphamide: 600 mg/m.sup.2/d) × 3d; (fludarabine: 30 mg/m.sup.2/d) × 4d Subjects are hospitalised for 7 days following infusion and monitored for safety, T-cell persistence, cytokine production with CT and MRI performed at weeks 4, 8, 16, 24 and 3 monthly thereafter until disease progression or early interventional withdrawal, long term follow up annually is planned for a 15 year period.

[0360] A subject will be considered completing the interventional phase of the study when he/she has received T-cell infusion and then progressed or died prior to disease progression. Optionally a second T-cell infusion may be given, and they will remain in the interventional phase of the study until they have further progression of disease. Once progression is established, no further efficacy assessments are performed other than overall survival. All subjects completing from the interventional portion of the study will enter the long-term follow-up (LTFU) phase for observation of delayed adverse events (AEs) during the 15 years post-infusion in accordance with FDA and EMA regulations. This study will be considered complete when the last living subject has completed LTFU.

[0361] To evaluate the safety and tolerability of MAGE-A4.sup.c1032T the incidence of dose limiting toxicities (DLTs) is monitored, determination is made of optimally tolerated dose range, adverse events (AEs), and Serious Adverse Events (SAEs); laboratory assessments, including chemistry, haematology, and coagulation; and cardiac assessments, including ECG and cardiac Troponin.

[0362] During the study MAGE-A4 is evaluated as a biomarker for tumour MAGE-A4 expression, and antitumor activity. This is performed to correlate the level of antigen expression in tumour level at Baseline, and post MAGE-A4.sup.c1032T cell infusion. Post-therapy MAGE-A4 expression in tumour over time is assessed to determine tumour immunity or resistance to MAGE-A4.sup.c1032T. Additionally, circulating cytokines were measured and evaluated for association with cytokine release syndrome (CRS) and other adverse events (AEs). Additionally post MAGE-A4.sup.c1032T cell infusion, transduced cell persistence is assessed by determination of serum level persistence of MAGE-A4.sup.c1032T engineered T-cell as measured by MAGE-A4.sup.c1032T vector copy number and MAGE-A4.sup.c1032T transduced T-cell number. Mean expression of specific surface markers on gene-modified T cells in subject blood and tumour were measured by fluorescence intensity. Killing profile and cytokine profile of genetically modified T cells were evaluated using flow cytometry in blood and tumor.

[0363] Biomarkers of subject sample including polymorphisms in cytokine genes and cytokine production.

[0364] To evaluate anti-tumour activity of MAGE-A4.sup.c1032T the following endpoints are monitored by RECIST v1.1; Overall Response Rate (ORR) defined as the proportion of subjects with a confirmed complete response (CR) or partial response (PR). Additional endpoints are monitored for duration of response (DoR), duration of stable disease (SD), progression free survival (PFS), overall survival (OS). Evaluation was made of the efficacy of the treatment by assessment of duration of response and assessment of overall survival. Also assessed were intervals for [0365] (a) the date of first T cell infusion dose and first documented evidence of CR or PR and evaluation of the efficacy of the treatment by assessment of time to first response. [0366] (b) the date of first documented evidence of CR or PR until first documented disease progression or death due to any cause. [0367] (c) the date of first documented evidence of stable disease (SD) until first documented disease progression or death due to any cause. [0368] (d) the date of first T cell infusion and the earliest date of disease, progression or death due to any cause. [0369] (e) between the date of first T cell infusion and date of death due to any cause.

[0370] Evaluation of the efficacy of the treatment by Number and % of subjects having any Long Term Follow Up Adverse Events (AEs), malignancy, neurologic disorder, rheumatologic or other autoimmune disorder, hematologic disorder, infections

[0371] Subjects were additionally monitored for safety and tolerability response through laboratory assessments including chemistry, hematology and coagulation, and anti-MAGE-A4 TCR antibodies, adverse events (AE), including serious adverse events (SAEs), dose limiting toxicities (DLT) NCI CTCAE and optimally tolerated dose range and evaluation of persistence of genetically modified T cells in the periphery and retention of heterologous TCR expression in the T cells PBMCs using PCR-based assay.

[0372] The same study was also extended to investigate treatment of subjects with ovarian cancer, melanoma and urothelial / bladder cancer and sarcoma (synovial sarcoma) with MAGE-A4.sup.c1032T for which data is presented below.

Results

[0373] Data presented in FIG. 1 provide the best overall response defined as the best response recorded from the date of T cell infusion until disease progression based on RECIST v1.1 criteria for responses in, head & neck cancer, and lung cancer for patients receiving a MAGE-A4.sup.c1032 T-cell infusion cell dose of between 35 × 10.sup.9 and 10 × 10.sup.9 cells (5 to 10 bn cells). These data support confirmed responses seen in subjects with head and neck cancer and lung cancer.

[0374] The data in FIG. 2 represent the CT scans of 42-yr-old male subject diagnosed age 25 and recently developed metastatic disease. The subject had moderate MAGE-A4 expression at baseline (16% 1+, 37% 2+, 41% 3+) and large disease burden; baseline SLD was 20 cm and was provided a first infusion of ~10 billion SPEAR MAGE-A4.sup.c1032 T-cells, adverse reaction was minimal developing grade 2 CRS (cytokine release syndrome) and cytopenias and consistent with those typically experienced by cancer patients undergoing cytotoxic chemotherapy and/or cancer immunotherapy. For lung cancer the tumour showed a greater than 45% decrease by RECIST 1.1 and symptoms of shortness of breath were resolved by treatment. The shortness of breath was due to fluid in pleural space, FIG. 2 lower left, seen resolved in lower right. FIG. 2 also shows a massive tumor displacing major blood vessels and compressing the right lung at baseline, upper left panel, and twelve weeks following treatment the tumor is dramatically decreased and non-target lesion absent, top right scan. FIG. 2 bottom right scan shows the lung expanded.

[0375] Data in FIG. 3 show the tumour responses for cohort subjects with different MAGE-A4 expressing cancers and the percent changes in sum of diameters in target lesions over a measured period of weeks as indicated following the date of T cell infusion. The data confirms the 24 week or 6 month efficacy of response in terms of tumour size reduction (36% reduction) for head and neck cancer treatment.

[0376] Data for ovarian cancer, urothelial / bladder cancer, and melanoma individual subjects also demonstrated a reduction in tumour size following treatment with MAGE-A4.sup.c1032 T-cell (between 5 to 10bn cell infusion) as determined by percentage changes in sum of diameters in target lesions based on the (SLD) sum of the long diameters for non-nodal lesions and short axis for nodal lesions; responses were evaluated by RECIST 1.1. Data demonstrated tumour SLD reductions of about 9% reduction for ovarian cancer at 12 weeks post infusion of MAGE-A4.sup.c1032T engineered T-cells, about 21% reduction for melanoma at 6 weeks post infusion of MAGE-A4.sup.c1032T engineered T-cells, approximately 67% reduction for urothelial / bladder cancer at 6 weeks post infusion of MAGE-A4.sup.c1032T engineered T-cells (FIG. 3). The data therefore demonstrates therapeutic efficacy and target tumor reductions observed in patients with ovarian, urothelial / bladder, and melanoma cancers.

[0377] In conclusion the data from the clinical studies show a confirmed response seen in subjects with head and neck cancer and lung cancer with tumor reductions also observed in patients with ovarian, bladder, and melanoma.

Sequences

[0378] SEQ ID NO: 1, MAGE A4

TABLE-US-00001 MSSEQKSQHC KPEEGVEAQE EALGLVGAQA PTTEEQEAAV SSSSPL VPGT LEEVPAAESA GPPQSPQGAS ALPTTISFTC WRQPNEGSSS Q EEEGPSTSP DAESLFREAL SNKVDELAHF LLRKYRAKEL VTKAEML ERV IKNYKRCFPV IFGKASESLK MIFGIDVKEV DPASNTYTLV TC LGLSYDGL LGNNQIFPKT GLLIIVLGTI AMEGDSASEE EIWEELGV MG VYDGREHTVY GEPRKLLTQD WVQENYLEYR QVPGSNPARY EFL WGPRALA ETSYVKVLEH VVRVNARVRI AYPSLREAAL LEEEEGV

[0379] SEQ ID NO: 2, MAGE A4 peptide

TABLE-US-00002 GVYDGREHTV

[0380] SEQ ID NO: 3; (CD8α)CDRs bold underlined, signal sequence italic underlined

TABLE-US-00003 MALPVTALLLPLALLLHAARPSQFRVSPLDRTWNLGETVELKCQVLLSNP TSGCSWLFQPRGAAASPTFLLYLSQNKPKAAEGLDTQRFSGKRLGDTFVL TLSDFRRENEGYYFCSALSNSIMYFSHFVPVFLPAKPTTTPAPRPPTPAP TIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSL VITLYCNHRNRRRVCKCPRPVVKSGDKPSLSARYV

[0381] SEQ ID NO: 4; (CD8α)

TABLE-US-00004 ATGGCCTTACCAGTGACCGCCTTGCTCCTGCCGCTGGCCTTGCTGCTCCA CGCCGCCAGGCCGAGCCAGTTCCGGGTGTCGCCGCTGGATCGGACCTGGA ACCTGGGCGAGACAGTGGAGCTGAAGTGCCAGGTGCTGCTGTCCAACCCG ACGTCGGGCTGCTCGTGGCTCTTCCAGCCGCGCGGCGCCGCCGCCAGTCC CACCTTCCTCCTATACCTCTCCCAAAACAAGCCCAAGGCGGCCGAGGGGC TGGACACCCAGCGGTTCTCGGGCAAGAGGTTGGGGGACACCTTCGTCCTC ACCCTGAGCGACTTCCGCCGAGAGAACGAGGGCTACTATTTCTGCTCGGC CCTGAGCAACTCCATCATGTACTTCAGCCACTTCGTGCCGGTCTTCCTGC CAGCGAAGCCCACCACGACGCCAGCGCCGCGACCACCAACACCGGCGCCC ACCATCGCGTCGCAGCCCCTGTCCCTGCGCCCAGAGGCGTGCCGGCCAGC GGCGGGGGGCGCAGTGCACACGAGGGGGCTGGACTTCGCCTGTGATATCT ACATCTGGGCGCCCTTGGCCGGGACTTGTGGGGTCCTTCTCCTGTCACTG GTTATCACCCTTTACTGCAACCACAGGAACCGAAGACGTGTTTGCAAATG TCCCCGGCCTGTGGTCAAATCGGGAGACAAGCCCAGCCTTTCGGCGAGAT ACGTCGGTTCAAGAGCTAAAAGAAGTGGTAGTGGTGCCCCTGTGA

[0382] SEQ ID NO: 5; (MAGE A4 TCR α chain) CDRs bold underlined

TABLE-US-00005 MKKHLTTFLVILWLYFYRGNGKNQVEQSPQSLIILEGKNCTLQCNYTVSP FSNLRWYKQDTGRGPVSLTILTFSENTKSNGRYTATLDADTKQSSLHITA SQLSDSASYICVVSGGTDSWGKLQFGAGTQVVVTPDIQNPDPAVYQLRDS KSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAW SNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNL SVIGFRILLLKVAGFNLLMTLRLWSSGSRAKR

[0383] SEQ ID NO: 6; (MAGE A4 TCR α chain coding sequence)

TABLE-US-00006 ATGAAGAAGCACCTGACCACCTTTCTCGTGATCCTGTGGCTGTACTTCTA CCGGGGCAACGGCAAGAACCAGGTGGAACAGAGCCCCCAGAGCCTGATCA TCCTGGAAGGCAAGAACTGCACCCTGCAGTGCAACTACACCGTGTCCCCC TTCAGCAACCTGCGGTGGTACAAGCAGGACACCGGCAGAGGCCCTGTGTC CCTGACCATCCTGACCTTCAGCGAGAACACCAAGAGCAACGGCCGGTACA CCGCCACCCTGGACGCCGATACAAAGCAGAGCAGCCTGCACATCACCGCC AGCCAGCTGAGCGATAGCGCCAGCTACATCTGCGTGGTGTCCGGCGGCAC AGACAGCTGGGGCAAGCTGCAGTTTGGCGCCGGAACACAGGTGGTCGTGA CCCCCGACATCCAGAACCCTGACCCTGCCGTGTACCAGCTGCGGGACAGC AAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGAC CAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGACCG TGCTGGACATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTGG TCCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATTAT CCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCTGCGACGTCAAGC TGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAGAACCTG AGCGTGATCGGCTTCAGAATCCTGCTGCTGAAGGTGGCCGGCTTCAACCT GCTGATGACCCTGAGACTGTGGTCCAGCGGCAGCCGGGCCAAGAGA

[0384] SEQ ID NO: 7; (MAGE A4 TCR β chain) CDRs bold underlined

TABLE-US-00007 MASLLFFCGAFYLLGTGSMDADVTQTPRNRITKTGKRIMLECSQTKGHDR MYWYRQDPGLGLRLIYYSFDVKDINKGEISDGYSVSRQAQAKFSLSLESA IPNQTALYFCATSGQGAYEEQFFGPGTRLTVLEDLKNVFPPEVAVFEPSE AEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQPA LNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVT QIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVL MAMVKRKDSRG

[0385] SEQ ID NO: 8; (MAGE A4 TCR β chain coding sequence)

TABLE-US-00008 ATGGCCAGCCTGCTGTTCTTCTGCGGCGCCTTCTACCTGCTGGGCACCGG CTCTATGGATGCCGACGTGACCCAGACCCCCCGGAACAGAATCACCAAGA CCGGCAAGCGGATCATGCTGGAATGCTCCCAGACCAAGGGCCACGACCGG ATGTACTGGTACAGACAGGACCCTGGCCTGGGCCTGCGGCTGATCTACTA CAGCTTCGACGTGAAGGACATCAACAAGGGCGAGATCAGCGACGGCTACA GCGTGTCCAGACAGGCTCAGGCCAAGTTCAGCCTGTCCCTGGAAAGCGCC ATCCCCAACCAGACCGCCCTGTACTTTTGTGCCACAAGCGGCCAGGGCGC CTACGAGGAGCAGTTCTTTGGCCCTGGCACCCGGCTGACAGTGCTGGAAG ATCTGAAGAACGTGTTCCCCCCAGAGGTGGCCGTGTTCGAGCCTTCTGAG GCCGAAATCAGCCACACCCAGAAAGCCACACTCGTGTGTCTGGCCACCGG CTTCTACCCCGACCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGG TGCACAGCGGCGTGTCCACCGATCCCCAGCCTCTGAAAGAACAGCCCGCC CTGAACGACAGCCGGTACTGCCTGAGCAGCAGACTGAGAGTGTCCGCCAC CTTCTGGCAGAACCCCAGAAACCACTTCAGATGCCAGGTGCAGTTTTACG GCCTGAGCGAGAACGACGAGTGGACCCAGGACAGAGCCAAGCCCGTGACA CAGATCGTGTCTGCCGAAGCTTGGGGGCGCGCCGATTGTGGCTTTACCAG CGAGAGCTACCAGCAGGGCGTGCTGAGCGCCACCATCCTGTACGAGATCC TGCTGGGAAAGGCCACACTGTACGCCGTGCTGGTGTCTGCCCTGGTGCTG ATGGCCATGGTCAAGCGGAAGGACAGCCGGGGC

[0386] SEQ ID NO: 9; (MAGE A4 TCR α chain variable region)136AA - CDRs bold underlined

TABLE-US-00009 MKKHLTTFLVILWLYFYRGNGKNQVEQSPQSLIILEGKNCTLQCNYTVSP FSNLRWYKQDTGRGPVSLTILTFSENTKSNGRYTATLDADTKQSSLHITA SQLSDSASYICVVSGGTDSWGKLQFGAGTQVVVTPD

[0387] SEQ ID NO: 10; (MAGE A4 TCR β chain variable region)133AA - CDRs bold underlined

TABLE-US-00010 MASLLFFCGAFYLLGTGSMDADVTQTPRNRITKTGKRIMLECSQTKGHDR MYWYRQDPGLGLRLIYYSFDVKDINKGEISDGYSVSRQAQAKFSLSLESA IPNQTALYFCATSGQGAYEEQFFGPGTRLTVLE

[0388] SEQ ID NO: 11; CDR1 MAGE A4 TCR α chain, (residues 48-53)

TABLE-US-00011 VSPFSN

[0389] SEQ ID NO: 12; CDR2 MAGE A4 TCR α chain, (residues 71-76)

TABLE-US-00012 LTFSEN

[0390] SEQ ID NO: 13; CDR3 MAGE A4 TCR α chain, (residues 111-125)

TABLE-US-00013 CVVSGGTDSWGKLQF

[0391] SEQ ID NO: 14; CDR1 MAGE A4 TCR β chain, (residues 46 - 50)

TABLE-US-00014 KGHDR

[0392] SEQ ID NO: 15; CDR2 MAGE A4 TCR β chain, (residues 68-73)

TABLE-US-00015 SFDVKD

[0393] SEQ ID NO: 16; CDR3 MAGE A4 TCR β chain, (residues 110 - 123)

TABLE-US-00016 CATSGQGAYEEQFF

[0394] SEQ ID NO: 17; CDR1 CD8α (residues 45-53)

TABLE-US-00017 VLLSNPTSG

[0395] SEQ ID NO: 18; CDR2 CD8α (residues 72-79)

TABLE-US-00018 YLSQNKPK

[0396] SEQ ID NO: 19; CDR3 CD8α (residues 118-123)

TABLE-US-00019 LSNSIM