Phase II Study of Short-Term Cultured Anti-Tumor Autologous Lymphocytes After Lymphocyte-Depleting Chemotherapy in Metastatic Melanoma

Study Overview
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Brief Summary
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Background: Most therapeutic therapies for metastatic melanoma have focused on the ability of T-cell lymphocytes to kill cells of tumors. An adaptive cell transfer therapy has been pioneered, in which cells are grown for a short time in the laboratory. The way they are grown may have a better effect in a patient's body than do other cells that are cultured for a longer time. Objectives: To determine whether tumor-infiltrating lymphocytes (TIL) 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 human leukocyte antigens (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 TIL cells (called young TIL cells) are 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 young TIL cells. Patients receive an IV infusion of the treated cells, followed by infusions the drug aldesleukin-2 (IL-2), 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 computed tomography (CT) 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: Tumor Infiltrating Lymphocytes (TIL) can mediate the regression of bulky metastatic melanoma when administered to an autologous patient with high dose (HD) IL-2 following a non-myeloablative (NMA) but lymphodepleting chemotherapy preparative regimen. Clinical investigations and preclinical animal models have demonstrated that less time in culture, longer telomeres, and a less differentiated lymphocyte phenotype are associated with TIL that are capable of mediating objective clinical responses and persisting long term in the host. Previous methods for generating TIL require screening for anti-tumor specificity using gamma-interferon (IFN) production by the TIL. However, in vitro screening depends on autologous tumor reagents that are often unavailable; and gamma-IFN release in vitro may not be the best correlate to in vivo efficacy. Additionally, this method necessitates long in vitro culture times (44 days), and therefore reduces the clonal heterogeneity of TIL cultures, and results in TIL cultures with shorter telomere lengths and phenotypes that are skewed toward a more differentiated phenotype. In Surgery Branch pre-clinical experiments, we evaluated a method for rapidly generating young TIL from melanoma tumors with optimal phenotypic characteristics. Objectives: In cohort 1, to determine the ability of autologous TIL cells infused after minimal in vitro culture in conjunction with high dose aldesleukin (IL-2) following a non-myeloablative lymphodepleting preparative regimen to mediate tumor regression in patients with metastatic melanoma. In cohort 2, to determine the ability of autologous cluster of differentiation 4 (CD4+) cell depleted TIL cells infused after minimal in vitro culture in conjunction with high dose aldesleukin (IL-2) following a non-myeloablative lymphodepleting preparative regimen to mediate tumor regression in patients with metastatic melanoma. In cohort 3, to determine the ability of autologous CD4+ cell depleted TIL cells infused after minimal in vitro culture in conjunction with high dose aldesleukin following chemoradiation lymphoid depleting regimen to mediate complete tumor regression in patients with metastatic melanoma. In a prospective randomized fashion, to compare the ability of autologous TIL cells (cohort 4) and autologous CD4plus cell depleted TIL cells (cohort regimen, to mediate tumor regression, progression free survival, and overall survival in patients with metastatic melanoma. Evaluate the toxicity of these treatment regimens. Determine the rate of repopulation of the young TIL cells in treated patients and establish in vitro correlates of TIL cultures that mediate objective response and in vivo persistence. Eligibility: Patients who 18 years of age or older must have: Metastatic melanoma; Normal values for basic laboratory values. Patients may not have: Received prior cell transfer therapy that included non-myeloablative or ablative chemotherapy; Concurrent major medical illnesses; Any form of immunodeficiency; Severe hypersensitivity to any of the agents used in this study; Contraindications for high dose IL-2 administration. Design: Patients will undergo resection to obtain tumor for generation of autologous TIL cultures. Cohort 1: All patients will receive a non-myeloablative lymphocyte depleting preparative regimen of cyclophosphamide (60 mg/kg/day IV) on days -7 and -6 and fludarabine (25 mg/m^2/day IV) on days -5 through -1. On day 0 patients will receive the infusion of autologous TIL and then begin high-dose aldesleukin (720,000 IU/kg IV every 8 hours for up to 15 doses). Clinical and Immunologic response will be evaluated about 4-6 weeks after TIL infusion. Using a small optimal two-stage Phase II design, initially 21 patients will be enrolled, and if two or more of the first 21 patients has a clinical response (partial response (PR) or complete response (CR)), accrual will continue to 41 patients, targeting a 20% goal for objective response. Cohort 1 will be closed with amendment D. Cohort 2 will be initiated with amendment D whereby CD4+ cells will be eliminated from the cultures, using the Miltenyi Clinimacs apparatus, prior to performing the rapid expansion of the young TIL cells. Patients in cohort 2 will receive CD4+ cell depleted young unselected TIL. Patients will also receive high dose IL-2 after non-myeloablative but lymphodepleting chemotherapy preparative regimen as described above for cohort 1. Clinical and immunologic response will be evaluated about 4-6 weeks after TIL infusion. Using a small optimal two-stage Phase II design, initially 18 patients will be enrolled, and if three or more of the first 18 patients have a clinical response (PR or CR), accrual will continue to 35 patients, targeting a 30% goal for objective response. With the initiation of Cohort 3 with amendment H, patients will only be accrued to Cohort 2 if they are not eligible to receive 600 cGy due to prior radiation, or to inability to mobilize cluster of differentiation 34 (CD34+) cells. Also at this time, accrual will be expanded to a total of 50 patients in cohort 2. Cohort 2 will be closed with amendment K. Cohort 3 will be initiated with amendment H, whereby patients will receive a chemoradiation lymphocyte depleting preparative regimen consisting of cyclophosphamide, fludarabine, and 600 cGy total body irradiation followed by intravenous infusion of autologous CD4+ cell depleted young TIL plus IV high dose IL-2. Clinical and immunologic response will be evaluated about 4-6 weeks after TIL infusion. Using a small optimal two-stage Phase II design, initially 26 patients will be enrolled, and if one or more of the first 26 patients have a complete response (CR), accrual will continue to 51 patients, targeting a 10% goal for complete response. Cohort 3 will be closed with amendment K. Prospective randomization between cohorts 4 and 5: Cohort 4: All patients will receive a non-myeloablative lymphocyte depleting preparative regimen of cyclophosphamide (60 mg/kg/day IV) on days -7 and -6 and fludarabine (25 mg/m^2/day IV) on days -5 through -1. On day 0 patients will receive the infusion of autologous TIL and then begin high-dose aldesleukin (720,000 IU/kg IV every 8 hours for up to 15 doses). Clinical and immunologic response will be evaluated about 4-6 weeks after TIL infusion. Cohort 5 All patients will receive a non-myeloablative lymphocyte depleting preparative regimen of cyclophosphamide (60 mg/kg/day IV) on days -7 and -6 and fludarabine (25 mg/m^2/day IV) on days -5 through -1. On day 0 patients will receive the infusion of autologous CD4+ depleted TIL and then begin high-dose aldesleukin (720,000 IU/kg IV every 8 hours for up to 15 doses). Clinical and immunologic response will be evaluated about 4-6 weeks after TIL infusion.

Official Title
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A Phase II Study Using Short-Term Cultured Anti-Tumor Autologous Lymphocytes Following a Lymphocyte Depleting Regimen in Metastatic Melanoma

Conditions
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Melanoma, Malignant Melanoma, Melanoma, Experimental, Experimental Melanomas

Intervention / Treatment
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* Biological: aldesleukin
* Biological: therapeutic autologous lymphocytes
* Drug: Cyclophosphamide
* Drug: Fludarabine phosphate
* Radiation: Total body irradiation


Participation Criteria
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Eligibility Criteria
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-INCLUSION CRITERIA: Measurable metastatic melanoma with at least one lesion that is resectable for tumor infiltrating lymphocytes (TIL) generation. Patients with one to three brain metastases are eligible (lesions greater than or equal to 1 cm each, or symptomatic lesions must have been treated and stable for 3 months). Greater than or equal to 18 years of age . Willing to practice birth control during treatment and for four months after receiving the preparative regimen. Life expectancy of greater than three months. Willing to sign a durable power of attorney. Able to understand and sign the Informed Consent Document. Clinical performance status of Eastern Cooperative Oncology Group (ECOG) 0 or 1. Hematology: Absolute neutrophil count greater than 1000/mm^3 without support of filgrastim. Normal white blood cell (WBC) (greater than 3000/ mm^3). Hemoglobin greater than 8.0 g/dl. Platelet count greater than 100,000/ mm^3. Serology: 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.) Seronegative for hepatitis B or hepatitis C. Chemistry: . Serum alanine aminotransferase (ALT)/aspartate aminotransferase (AST) less than three times the upper limit of normal. Serum creatinine less than or equal to 1.6 mg/dl. Total bilirubin less than or equal to 2 mg/dl, except in patients with Gilbert's Syndrome who must have a total bilirubin less than 3 mg/dl. 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). Patients may have undergone minor surgical procedures with the past 3 weeks, as long as all toxicities have recovered to grade 1 or less or as specified in the eligibility criteria in Section 2.1.1. Six weeks must have elapsed since prior MDX-010 (Ipilimumab) therapy to allow antibody levels to decline. Patients who have previously received any anti-CTLA4 (cytotoxic T-lymphocyte antigen 4) antibody and experienced treatment related colitis must have a normal colonoscopy with normal colonic biopsies. EXCLUSION CRITERIA: Prior cell transfer therapy that included non-myeloablative or ablative chemotherapy (for cohorts 4 and 5). 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. Systemic steroid therapy required. Active systemic infections, coagulation disorders or other active major medical illnesses of the cardiovascular, respiratory or immune system, as evidenced by a positive stress thallium or comparable test, myocardial infarction, cardiac arrhythmias, obstructive or restrictive pulmonary disease. Any form of primary immunodeficiency (such as Severe Combined Immunodeficiency Disease and Acquired Immune Deficiency Syndrome (AIDS)). 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.) History of severe immediate hypersensitivity reaction to any of the agents used in this study. History of coronary revascularization or ischemic symptoms. Any patient known to have an left ventricular ejection fraction (LVEF) less than or equal to 45%. Documented LVEF of less than or equal to 45% 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. Documented forced expiratory volume 1 (FEV1) less than or equal to 60% predicted tested in patients with: A prolonged history of cigarette smoking Symptoms of respiratory dysfunction

Ages Eligible for Study
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Minimum Age: 18 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Allocation: Non-Randomized
Intervention Model: Single Group Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Cohort 1 - NMA, TIL, aldesleukin<br>Cohort 1 - Nonmyeloablative (NMA), tumor infiltrating lymphocytes (TIL), & high dose (HD) aldesleukin: Nonmyeloablative chemotherapeutic conditioning regimen followed by bulk young tumor infiltrating lymphocytes and high dose aldesleukin. Cyclophosphamide 60 mg/kg intravenous (IV) daily x 2 days. Fludarabine 25 mg/m^2 intravenous (IV) daily x 5 days. Bulk young TIL Aldesleukin 720,000 IU/kg intravenous (IV) (based on body weight) over 15 minutes every eight hours for up to 5 days. Cohort 1 = unselected TIL | Biological: aldesleukin<br>* Given subcutaneously every 8 hours for up to 15 doses, day 0, 720,000 IU/kg<br>* Other names: IL-2;Biological: therapeutic autologous lymphocytes<br>* Given as infusion, up to 3 x 10^11 lymphocytes (minimum of 1 x 10^9), day 0<br>Drug: Cyclophosphamide<br>* Given intravenously 60 mg/kg/day, day -7 to -6<br>Drug: Fludarabine phosphate<br>* Given intravenously 25 mg/m^2/day over 15-30 minutes, day -5 to -1<br>* Other names: Fludara;|
| Experimental: Cohort 2 - NMA, CD4+ TIL, aldesleukin<br>Cohort 2 - Nonmyeloablative (NMA), cluster of differentiation 4 (CD4+) depleted tumor infiltrating lymphocytes (TIL), aldesleukin: Nonmyeloablative chemotherapeutic conditioning regimen followed by CD4+ depleted tumor infiltrating lymphocytes and high dose (HD) aldesleukin. Cyclophosphamide 60 mg/kg intravenous (IV) daily x 2 days. Fludarabine 25 mg/m^2 intravenous (IV) daily x 5 days. CD4+ depleted TIL Aldesleukin 720,000 IU/kg intravenous (IV) (based on body weight) over 15 minutes every eight hours for up to 5 days. Cohort 2 = CD4+ depleted (selected) TIL | Biological: aldesleukin<br>* Given subcutaneously every 8 hours for up to 15 doses, day 0, 720,000 IU/kg<br>* Other names: IL-2;Biological: therapeutic autologous lymphocytes<br>* Given as infusion, up to 3 x 10^11 lymphocytes (minimum of 1 x 10^9), day 0<br>Drug: Cyclophosphamide<br>* Given intravenously 60 mg/kg/day, day -7 to -6<br>Drug: Fludarabine phosphate<br>* Given intravenously 25 mg/m^2/day over 15-30 minutes, day -5 to -1<br>* Other names: Fludara;|
| Experimental: Cohort 3 - NMA, total body irradiation<br>Cohort 3 - Nonmyeloablative (NMA), total body irradiation (TBI): Nonmyeloablative chemotherapeutic conditioning regimen and 2 gray units (Gy) of total body irradiation followed by cluster of differentiation 4 (CD4+) depleted tumor infiltrating lymphocytes and high dose (HD) aldesleukin. Cyclophosphamide 60 mg/kg intravenous (IV) daily x 2 days. Fludarabine 25 mg/m^2 intravenous (IV) daily x 5 days. CD4+ depleted TIL 2Gy (gray units) of total body irradiation (TBI) twice on day -2 and once on day -1 (total dose 6 Gy) at a rate of 0.07 Gy/minute using a linear accelerator in Radiation Oncology Aldesleukin 720,000 IU/kg intravenous (IV) (based on body weight) over 15 minutes every eight hours for up to 5 days. Cohort 3 = CD4 + depleted (selected) TIL + 600Gy radiation | Biological: aldesleukin<br>* Given subcutaneously every 8 hours for up to 15 doses, day 0, 720,000 IU/kg<br>* Other names: IL-2;Biological: therapeutic autologous lymphocytes<br>* Given as infusion, up to 3 x 10^11 lymphocytes (minimum of 1 x 10^9), day 0<br>Drug: Cyclophosphamide<br>* Given intravenously 60 mg/kg/day, day -7 to -6<br>Drug: Fludarabine phosphate<br>* Given intravenously 25 mg/m^2/day over 15-30 minutes, day -5 to -1<br>* Other names: Fludara;Radiation: Total body irradiation<br>* 600 cGy<br>* Other names: TBI;|
| Experimental: Cohort 4 - NMA, young TIL, aldesleukin<br>Cohort 4 - Nonmyeloablative (NMA), tumor infiltrating lymphocytes (TIL), aldesleukin: Nonmyeloablative chemotherapeutic conditioning regimen followed by bulk young tumor infiltrating lymphocytes and high dose (HD) aldesleukin. Cyclophosphamide 60 mg/kg intravenous (IV) daily x 2 days. Fludarabine 25 mg/m^2 intravenous (IV) daily x 5 days. Bulk young TIL Aldesleukin 720,000 IU/kg intravenous (IV) (based on body weight) over 15 minutes every eight hours for up to 5 days. Cohort 4 = unselected TIL - it is the SAME as cohort 1 | Biological: aldesleukin<br>* Given subcutaneously every 8 hours for up to 15 doses, day 0, 720,000 IU/kg<br>* Other names: IL-2;Biological: therapeutic autologous lymphocytes<br>* Given as infusion, up to 3 x 10^11 lymphocytes (minimum of 1 x 10^9), day 0<br>Drug: Cyclophosphamide<br>* Given intravenously 60 mg/kg/day, day -7 to -6<br>Drug: Fludarabine phosphate<br>* Given intravenously 25 mg/m^2/day over 15-30 minutes, day -5 to -1<br>* Other names: Fludara;|
| Experimental: Cohort 5 - NMA, CD4+TIL, HD aldesleukin<br>Cohort 5 - Nonmyeloablative (NMA), cluster of differentiation 4 (CD4+) tumor infiltrating lymphocytes (TIL), high dose (HD) aldesleukin: Nonmyeloablative chemotherapeutic conditioning regimen followed by CD4+ depleted tumor infiltrating lymphocytes and high dose aldesleukin. Cyclophosphamide 60 mg/kg intravenous (IV) daily x 2 days. Fludarabine 25 mg/m^2 intravenous (IV) daily x 5 days. CD4+ depleted TIL Aldesleukin 720,000 IU/kg intravenous (IV) (based on body weight) over 15 minutes every eight hours for up to 5 days. Cohort 5 = CD4 + depleted TIL - it is the SAME as cohort 2 | Biological: aldesleukin<br>* Given subcutaneously every 8 hours for up to 15 doses, day 0, 720,000 IU/kg<br>* Other names: IL-2;Biological: therapeutic autologous lymphocytes<br>* Given as infusion, up to 3 x 10^11 lymphocytes (minimum of 1 x 10^9), day 0<br>Drug: Cyclophosphamide<br>* Given intravenously 60 mg/kg/day, day -7 to -6<br>Drug: Fludarabine phosphate<br>* Given intravenously 25 mg/m^2/day over 15-30 minutes, day -5 to -1<br>* Other names: Fludara;|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Clinical Response | Clinical response is defined as complete response (CR)- a disappearance of all target lesions, partial response (PR) - at least a 30% decrease in the sum of the longest diameter (LD) of target lesions taking as reference the baseline sum LD. Progression (PD)- at least a 20% increase in the sum of the LD of target lesions taking as reference the smallest sum LD recorded since the treatment started or the appearance of one or more new lesions. Stable disease (SD) - neither sufficient shrinkage to qualify for PR nor sufficient increase to qualify for PD taking as references the smallest sum LD. | every 1-3 months until disease progression. Total length of time -8/7/2007 to 9/27/2012 | 
| Toxicity | Here is the number of participants with adverse events. For a detailed list of adverse events, see the adverse event module. | 5 years | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Clinical Response, Immunotherapy, Cancer, Cytokines, Adoptive Cell Therapy, Melanoma, Skin Cancer, Malignant Melanoma