Phase II Study of Metastatic Melanoma With Lymphodepleting Conditioning and Infusion of Anti-MART-1 F5 TCR-Gene-Engineered Lymphocytes

NCT ID: NCT00509288

Last Updated: 2012-12-28

Study Results

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Basic Information

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Recruitment Status

COMPLETED

Clinical Phase

PHASE2

Total Enrollment

24 participants

Study Classification

INTERVENTIONAL

Study Start Date

2007-06-30

Study Completion Date

2012-07-31

Brief Summary

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Background:

* Human peripheral blood lymphocytes have been engineered to express a T-cell receptor (TCR) that recognizes a blood type,HLA-A 0201 (human leukocyte antigen) derived from the gp100 protein. A retroviral vector was constructed that can deliver the T-cell receptor (TCR) to cells.
* Patients' cells will be converted into cells able to recognize and fight melanoma tumors.

Objectives:

* To determine whether TCR-engineered lymphocytes can be put in cells removed from patients' tumors or blood and then reinfused, with the purpose of shrinking tumors.
* To evaluate safety and effectiveness of the treatment.

Eligibility:

* Patients 18 years of age or older with metastatic cancer melanoma (cancer that has spread beyond the original site).
* Patient's leukocyte antigen type is HLA-A 0201.

Design:

-Patients undergo the following procedures:

* Leukapheresis (on two occasions). This is a method of collecting large numbers of white blood cells. The cells obtained in the first leukapheresis procedure are grown in the laboratory, and the anti-MART-1 protein is inserted into the cells using an inactivated (harmless) virus in a process called retroviral transduction. Cells collected in the second leukapheresis procedure are used to evaluate the effectiveness of the study treatment.
* Chemotherapy. Patients are given chemotherapy through a vein (intravenously, IV) over 1 hour for 2 days to suppress the immune system so that the patient's immune cells do not interfere with the treatment.
* Treatment with anti-melanoma antigen recognized by T-cells (MART)-1. Patients receive an intravenous (IV) infusion of the treated cells containing anti-MART-1 protein, followed by infusions of a drug called IL-2 (aldesleukin), which helps boost the effectiveness of the treated white cells.
* Patients are given support medications to prevent complications such as infections.
* Patients may undergo a tumor biopsy (removal of a small piece of tumor tissue).
* Patients are evaluated with laboratory tests and imaging tests, such as CT (computed tomography) scans, 4 to 6 weeks after treatment and then once a month for 3 to 4 months to determine the response to treatment.
* Patients have blood tests at 3, 6, and 12 months and then annually for 5 years.

Detailed Description

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Background:

* We have engineered human TIL (tumor infiltrating lymphocytes) and peripheral blood lymphocytes (PBLs) to express an anti-MART-1 T-cell receptor that recognizes an HLA-A\*0201 restricted epitope derived from the TIL clone DMF5.
* We constructed a single retroviral vector that contains both alpha and beta chains and can mediate genetic transfer of this TCR with high efficiency without the need to perform any selection.
* In co-cultures with HLA-A\*0201 positive melanoma, anti-MART-1 F5 TCR transduced T cells secreted significant amount of interferon (IFN)-gamma (but no significant secretion was observed in control co-cultures with cell lines.
* The anti-MART-1 F5 TCR transduced T-cells could efficiently kill HLA-A\*0201 positive tumors. There was little or no recognition of normal fibroblasts cells.
* This TCR is over 10 times more reactive with melanoma cells than the MART-1 TCR that mediated tumor regression in two patients with metastatic melanoma.

Objectives:

Primary objectives:

-Determine if the administration of anti-MART-1 F5 TCR -engineered peripheral blood lymphocytes (PBL) or tumor infiltrating lymphocytes (TIL) and aldesleukin to patients following a nonmyeloablative but lymphoid depleting preparative regimen will result in clinical tumor regression in patients with metastatic melanoma.

Secondary objectives:

* Determine the in vivo survival of TCR gene-engineered cells.
* Determine the toxicity profile of this treatment regimen.

Eligibility:

Patients who are HLA-A 0201 positive and 18 years of age or older must have:

* metastatic melanoma;
* previously received and have been a non-responder to or recurred after aldesleukin;
* normal values for basic laboratory values.

Patients may not have:

* concurrent major medical illnesses;
* any form of primary or secondary immunodeficiency;
* severe hypersensitivity to any of the agents used in this study;
* contraindications for high dose aldesleukin administration.

Design:

* If TIL can be obtained and grown, but are non-reactive, patients will be assigned to receive TIL transduced with the anti-MART-1 F5 TCR retroviral vector. If TIL cannot be obtained, PBMC will be obtained by leukapheresis (approximately 5 X 10(9) cells) and cultured in the presence of anti-CD3 (OKT3) and aldesleukin and transduced with the anti-MART-1 F5 TCR retroviral vector. If TIL cells are reactive to autologous tumor or major histocompatibility complex (MHC)-matched tumor cells or PBL cannot be grown, patients will not be treated on this protocol.
* Transduction is initiated by exposure of approximately 10(8) to 5 times 10(8) cells to retroviral vector supernatant containing the anti-MART-1 F5 TCR genes. These transduced cells will be expanded and tested for their anti-tumor activity.
* Once engineered lymphocytes are demonstrated to be biologically active according to the strict criteria outlined in the Certificate of Analysis, patients will receive a nonmyeloablative but lymphocyte depleting preparative regimen consisting of cyclophosphamide and fludarabine followed by intravenous infusion of ex vivo tumor reactive, TCR gene transduced cells plus IV aldesleukin (720,000 IU/kg q8h for a maximum of 15 doses).
* Patients will undergo complete evaluation of tumor with physical examination, CT of the chest, abdomen and pelvis and clinical laboratory evaluation four to six weeks after treatment and then monthly for approximately 3 to 4 months or until off study criteria are met.
* The study will be conducted using a phase II optimal design where initially 21 evaluable patients will be enrolled into each of two cohorts. If 0 or 1 of the 21 patients per cohort experiences a clinical response, then no further patients will be enrolled but if 2 or more of the first 21 evaluable patients enrolled in that cohort have a clinical response, then accrual to that cohort will continue until a total of 41 evaluable patients have been enrolled in that cohort.
* The objective will be to determine in two cohorts if the combination of high dose aldesleukin, lymphocyte depleting chemotherapy, and anti-MART-1 F5 TCR-gene engineered lymphocytes (TIL and PBL) is able to be associated with a clinical response rate that can rule out 5% (p0=0.05) in favor of a modest 20% PR (partial response) + CR (complete response) rate (p1=0.20).

Conditions

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Melanoma Skin Cancer

Keywords

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Refractory Gene Therapy HLA-A2 Positive Stage IV Melanoma Melanoma Skin Cancer Malignant Melanoma

Study Design

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Allocation Method

NON_RANDOMIZED

Intervention Model

SINGLE_GROUP

Primary Study Purpose

TREATMENT

Blinding Strategy

NONE

Study Groups

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anti-MART-1 F5 TCR PBL + HD IL-2

Patients treated with peripheral blood lymphocytes (PBL)

Group Type EXPERIMENTAL

autologous anti-MART-1 F5 T-cell receptor

Intervention Type BIOLOGICAL

Autologous anti-MART-1 F5 T-cell receptor gene-engineered tumor infiltrating lymphocytes.

A minimum of approximately 5 X 10\^8 cells will be given up to 3x10\^11 anti-MART-1 F5 TCR engineered TIL or PBL.

Cyclophosphamide

Intervention Type DRUG

Day -7 to -5: Cyclophosphamide 60 mg/kg/day X 2 days intravenous (IV) in 250 ml D5W with mesna 15 mg/kg/day X 2 days over 1 hr

Fludarabine

Intervention Type DRUG

Day -5 to 1: Fludarabine 25 mg/m\^2/day intravenous piggy back (IVPB) daily over 30 minutes for 5 days

Aldesleukin

Intervention Type BIOLOGICAL

Day 0: Cells will be infused intravenously (i.v.). Patients will receive up to 3x10e\^11 (with a minimum of 5x10e\^8 cells) anti-MART-1 F5 TCR engineered TIL or PBL Aldesleukin (based on total body weight) 720,000 IU/kg intravenous (IV) over 15 minute every eight hours beginning within 24 hours of cell infusion

anti-MART-1 F5 TCR TIL + HD IL-2

Patients treated with TIL (tumor infiltrating lymphocytes).

Group Type EXPERIMENTAL

Cyclophosphamide

Intervention Type DRUG

Day -7 to -5: Cyclophosphamide 60 mg/kg/day X 2 days intravenous (IV) in 250 ml D5W with mesna 15 mg/kg/day X 2 days over 1 hr

Fludarabine

Intervention Type DRUG

Day -5 to 1: Fludarabine 25 mg/m\^2/day intravenous piggy back (IVPB) daily over 30 minutes for 5 days

Aldesleukin

Intervention Type BIOLOGICAL

Day 0: Cells will be infused intravenously (i.v.). Patients will receive up to 3x10e\^11 (with a minimum of 5x10e\^8 cells) anti-MART-1 F5 TCR engineered TIL or PBL Aldesleukin (based on total body weight) 720,000 IU/kg intravenous (IV) over 15 minute every eight hours beginning within 24 hours of cell infusion

autologous anti-MART-1 F5 T-cell receptor gene-engineered tumor infiltrating lymphocytes

Intervention Type BIOLOGICAL

Interventions

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autologous anti-MART-1 F5 T-cell receptor

Autologous anti-MART-1 F5 T-cell receptor gene-engineered tumor infiltrating lymphocytes.

A minimum of approximately 5 X 10\^8 cells will be given up to 3x10\^11 anti-MART-1 F5 TCR engineered TIL or PBL.

Intervention Type BIOLOGICAL

Cyclophosphamide

Day -7 to -5: Cyclophosphamide 60 mg/kg/day X 2 days intravenous (IV) in 250 ml D5W with mesna 15 mg/kg/day X 2 days over 1 hr

Intervention Type DRUG

Fludarabine

Day -5 to 1: Fludarabine 25 mg/m\^2/day intravenous piggy back (IVPB) daily over 30 minutes for 5 days

Intervention Type DRUG

Aldesleukin

Day 0: Cells will be infused intravenously (i.v.). Patients will receive up to 3x10e\^11 (with a minimum of 5x10e\^8 cells) anti-MART-1 F5 TCR engineered TIL or PBL Aldesleukin (based on total body weight) 720,000 IU/kg intravenous (IV) over 15 minute every eight hours beginning within 24 hours of cell infusion

Intervention Type BIOLOGICAL

autologous anti-MART-1 F5 T-cell receptor gene-engineered tumor infiltrating lymphocytes

Intervention Type BIOLOGICAL

Other Intervention Names

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Cytoxan Fludara IL-2

Eligibility Criteria

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Inclusion Criteria

1. Metastatic melanoma with measurable disease
2. Previously received high dose IL-2 (aldesleukin) and have been either non-responders (progressive disease) or have recurred.
3. Positive for MART-1 by immunohistochemistry (IHC)
4. Greater than or equal to 18 years of age.
5. Willing to sign a durable power of attorney
6. Able to understand and sign the Informed Consent Document
7. Clinical performance status of Eastern Cooperative Oncology Group (ECOG) 0 or 1.
8. Life expectancy of greater than three months.
9. Patients of both genders must be willing to practice birth control for four months after receiving the preparative regimen.
10. Patients must be human leukocyte antigen (HLA-A)\*0201 positive
11. Serology:

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1. Seronegative for human immunodeficiency virus (HIV) antibody. (The experimental treatment being evaluated in this protocol depends on an intact immune system. Patients who are HIV seropositive can have decreased immune -competence and thus be less responsive to the experimental treatment and more susceptible to its toxicities.)
2. Seronegative for hepatitis B antigen and hepatitis C antibody unless antigen negative.
3. Women of child-bearing potential must have a negative pregnancy test because of the potentially dangerous effects of the preparative chemotherapy on the fetus.

l. Hematology:
1. Absolute neutrophil count greater than 1000/mm\^3 without the support of filgrastim.
2. White blood cell (WBC) (greater than 3000/mm\^3).
3. Platelet count greater than 100,000/ mm\^3.
4. Hemoglobin greater than 8.0 g/dl.

m. Chemistry:
1. Serum alanine aminotransferase (ALT)/aspartate aminotransferase (AST) less or equal to 2.5 times the upper limit of normal.
2. Serum creatinine less than or equal to 1.6 mg/dl.
3. Total bilirubin less than or equal to 1.5 mg/dl, except in patients with Gilbert's Syndrome who must have a total bilirubin less than 3.0 mg/dl.

n. More than four weeks must have elapsed since any prior systemic therapy at the time the patient receives the preparative regimen, and patients' toxicities must have recovered to a grade 1 or less (except for toxicities such as alopecia or vitiligo).

o. Six weeks must have elapsed since prior anti-CTLA4 antibody therapy to allow antibody levels to decline.

p. Patients who have previously received anti-CTLA4 antibody must have a normal colonoscopy with normal colonic biopsies.

Exclusion Criteria

1. Patients with reactive TIL (IFN-gamma release greater than 200 pg/mL) available based on overnight co-culture assay with autologous tumor or MHC-matched tumor cells.
2. Women of child-bearing potential who are pregnant or breastfeeding because of the potentially dangerous effects of the preparative chemotherapy on the fetus or infant.
3. Active systemic infections, coagulation disorders or other major medical illnesses of the cardiovascular, respiratory or immune system, myocardial infarction, cardiac arrhythmias, obstructive or restrictive pulmonary disease.
4. Any form of primary immunodeficiency (such as Severe Combined Immunodeficiency Disease).
5. Opportunistic infections (The experimental treatment being evaluated in this protocol depends on an intact immune system. Patients who have decreased immune competence may be less responsive to the experimental treatment and more susceptible to its toxicities).
6. Systemic steroid therapy.
7. History of severe immediate hypersensitivity reaction to any of the agents used in this study.
8. History of coronary revascularization or ischemic symptoms.
9. Any patient known to have an left ventricular ejection fraction (LVEF) less than or equal to 45 percent.
10. Documented LVEF of less than or equal to 45 percent tested in patients with:

* Clinically significant atrial and/or ventricular arrhythmias including but not limited to: atrial fibrillation, ventricular tachycardia, second or third degree heart block.
* Age greater than or equal to 60 years old.
11. Documented forced expiratory volume 1 (FEV1) less than or equal to 60 percent predicted tested in patients with:

* A prolonged history of cigarette smoking (20 pk/yrs of smoking).
* Symptoms of respiratory dysfunction.
Minimum Eligible Age

18 Years

Eligible Sex

ALL

Accepts Healthy Volunteers

No

Sponsors

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National Cancer Institute (NCI)

NIH

Sponsor Role lead

Responsible Party

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National Institutes of Health

Principal Investigators

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Steven Rosenberg, M.D.

Role: PRINCIPAL_INVESTIGATOR

National Cancer Institute, National Institutes of Health

Locations

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National Institutes of Health Clinical Center, 9000 Rockville Pike

Bethesda, Maryland, United States

Site Status

Countries

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United States

References

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Kawakami Y, Eliyahu S, Delgado CH, Robbins PF, Rivoltini L, Topalian SL, Miki T, Rosenberg SA. Cloning of the gene coding for a shared human melanoma antigen recognized by autologous T cells infiltrating into tumor. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3515-9. doi: 10.1073/pnas.91.9.3515.

Reference Type BACKGROUND
PMID: 8170938 (View on PubMed)

Kawakami Y, Eliyahu S, Sakaguchi K, Robbins PF, Rivoltini L, Yannelli JR, Appella E, Rosenberg SA. Identification of the immunodominant peptides of the MART-1 human melanoma antigen recognized by the majority of HLA-A2-restricted tumor infiltrating lymphocytes. J Exp Med. 1994 Jul 1;180(1):347-52. doi: 10.1084/jem.180.1.347.

Reference Type BACKGROUND
PMID: 7516411 (View on PubMed)

Kawakami Y, Eliyahu S, Delgado CH, Robbins PF, Sakaguchi K, Appella E, Yannelli JR, Adema GJ, Miki T, Rosenberg SA. Identification of a human melanoma antigen recognized by tumor-infiltrating lymphocytes associated with in vivo tumor rejection. Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6458-62. doi: 10.1073/pnas.91.14.6458.

Reference Type BACKGROUND
PMID: 8022805 (View on PubMed)

Abate-Daga D, Hanada K, Davis JL, Yang JC, Rosenberg SA, Morgan RA. Expression profiling of TCR-engineered T cells demonstrates overexpression of multiple inhibitory receptors in persisting lymphocytes. Blood. 2013 Aug 22;122(8):1399-410. doi: 10.1182/blood-2013-04-495531. Epub 2013 Jul 16.

Reference Type DERIVED
PMID: 23861247 (View on PubMed)

Related Links

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Other Identifiers

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07-C-0175

Identifier Type: -

Identifier Source: secondary_id

070175

Identifier Type: -

Identifier Source: org_study_id