Dendritic Cells (White Blood Cells) Vaccination for Advanced Melanoma

Study Overview
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Brief Summary
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The purpose of this study is to investigate a method of using dendritic cells (a kind of white blood cell) as a vaccine to stimulate your own immune system to react to your melanoma cells.

Detailed Description
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Eligible patients that provide written informed consent will undergo apheresis to collect blood mononuclear cells for vaccine production. All patients will be given cyclophosphamide 300mg/m2 IV three days prior to vaccine dose #1 in order to deplete regulatory T cells. All patients will receive mature DC for each dose of vaccine. For each dose all patients will receive autologous dendritic cells pulsed with 2 gp100 melanoma peptides (G209-2M and G280-9V) plus up to an additional 10 unique melanoma tumor-specific peptides. All patients will receive booster doses with mature DC. The DC vaccine will be given intravenously every three weeks for a total of six vaccine doses. Peripheral blood (16 ml) will be taken weekly to monitor the immune response to each peptide by tetramer assay. Apheresis is repeated after vaccine dose #3 and dose #6 in order to collect PBMC for immune monitoring. Restaging is performed after three and six vaccine doses. Patients with stable disease or better (partial response/complete response) after six doses will be eligible to receive additional vaccinations as maintenance therapy every 2 months until progression.

Official Title
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Mature Dendritic Cell Vaccination Against gp100 in Patients With Advanced Melanoma

Conditions
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Melanoma

Intervention / Treatment
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* Drug: cyclophosphamide
* Biological: Mature dendritic cell vaccine


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Unresectable stage III and stage IV M1a/M1b/M1c melanoma including patients with uveal melanoma Age ≥ 18 years Life expectancy ≥ 4 months ECOG performance status 0-2 At least 28 days from prior treatment (including adjuvant interferon) except in cases of a BRAF inhibitor (such as vemurafenib); concurrent treatment with a BRAF inhibitor +/- MEK inhibitor is permitted Required initial laboratory values (submitted within 14 days prior to registration): WBC >3,000/mm3 Hg ≥ 9.0 gm/dl Platelets >75,000/mm3 Serum Bilirubin < 2.0 mg/dl Serum Creatinine < 2.0 mg/dl Sexually active women of childbearing potential must use effective birth control during the trial and for at least two months following the trial, and sexually active men must be willing to avoid fathering a new child while receiving therapy. Exclusion Criteria: Prior treatment with more than one line of cytotoxic chemotherapy; prior treatment with one line of cytotoxic chemotherapy is permitted. Prior treatment with targeted therapy (such as ipilumumab, anti-PD1, and BRAF inhibitor) is permitted. Active untreated CNS metastasis Active infection Prior malignancy (except non-melanoma skin cancer) within 3 years Pregnant or nursing Concurrent treatment with corticosteroids; local (inhaled or topical) steroids are permitted. Inability to provide adequate informed consent Known allergy to eggs Prior history or uveitis or autoimmune inflammatory eye disease. Known positivity for hepatitis BsAg, hepatitis C antibody, or HIV antibody.

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: Parallel Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Dendritic Cell Vaccine (First Group)<br>Blood mononuclear cells will be collected for vaccine production through apheresis. Patients will be given cyclophosphamide 300mg/m2 IV 3 days prior to vaccine dose #1 in order to deplete regulatory T cells. Patients will receive mature DC for each dose of vaccine and will receive autologous dendritic cells. The DC vaccine will be given intravenously every 3 weeks for a total of 6 doses. Peripheral blood will be taken weekly to monitor the immune response. Apheresis is repeated after vaccine dose #3 and dose #6 in order to collect PBMC for immune monitoring. Patients with stable disease or better after 6 doses will be eligible to receive additional vaccinations as maintenance therapy every 2 months until progression. | Drug: cyclophosphamide<br> <br> * Other names: Cytoxan;Biological: Mature dendritic cell vaccine<br> <br> |
| Experimental: Dendritic Cell Vaccine (Second Group)<br>Blood mononuclear cells will be collected for vaccine production through apheresis. Patients will be given cyclophosphamide 300mg/m2 IV 3 days prior to vaccine dose #1 in order to deplete regulatory T cells. Patients will receive mature DC for each dose of vaccine and will receive autologous dendritic cells. The DC vaccine will be given intravenously every 3 weeks for a total of 6 doses. Peripheral blood will be taken weekly to monitor the immune response. Apheresis is repeated after vaccine dose #3 and dose #6 in order to collect PBMC for immune monitoring. Patients with stable disease or better after 6 doses will be eligible to receive additional vaccinations as maintenance therapy every 2 months until progression. | Drug: cyclophosphamide<br> <br> * Other names: Cytoxan;Biological: Mature dendritic cell vaccine<br> <br> |
| Experimental: Dendritic Cell Vaccine (Third Group)<br>Blood mononuclear cells will be collected for vaccine production through apheresis. Patients will be given cyclophosphamide 300mg/m2 IV 3 days prior to vaccine dose #1 in order to deplete regulatory T cells. Patients will receive mature DC for each dose of vaccine and will receive autologous dendritic cells. The DC vaccine will be given intravenously every 6 weeks for a total of 3 doses. Peripheral blood will be taken weekly to monitor the immune response. Apheresis is repeated after vaccine dose #3 in order to collect PBMC for immune monitoring. | Drug: cyclophosphamide<br> <br> * Other names: Cytoxan;Biological: Mature dendritic cell vaccine<br> <br> |

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Immunological response based on measuring increased numbers of peptide specific CD8+ T cells as calculated by the tetramer assay. | Starting on Day 0, two tubes will be drawn weekly until Day 64. Thereafter, two tubes will be drawn every 21 days until Day 190. For patients receiving maintenance treatment, blood is drawn every month. Data are presented as the percentage of CD8+ T cells positive for tetramer binding based on gating variables set using the iMASC reagent kit (Beckman Coulter). | Through completion of treatment | 
| Safety and tolerability of the mature dendritic cell vaccine as measured by adverse events | The descriptions and grading scales found in the revised NCI Common Terminology Criteria for Adverse Events (CTCAE) version 3.0 will be utilized for all toxicity reporting. | 30 days after end of treatment | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Time to progression |  | Through completion of treatment or until progressive disease | 
| Regulatory T cell depletion after cyclophosphamide administration. | Regulatory T cells (Treg) are defined as CD4+CD25+foxP3+ (triple positive) cells. At the indicated time points, the percentage of Treg cells is determined by 3 color flow cytometry. The depletion of Treg is defined as follows [Treg baseline - Treg nadir/ Treg baseline x 100= % depletion]. | Day -3 (72 hours prior to vaccine dose 1) | 
| Safety and side effect profile of mDC administered to patients given after a single dose of cyclophosphamide. |  | Day 0 (prior to vaccine dose 1) | 
| Clinical response rate using RECIST criteria |  | After third vaccine, sixth vaccine, and then every 8 weeks |