Peptide Vaccination Associated With Tumoral Immunomodulation in Patients With Advanced Metastatic Melanoma
Peptide Vaccination Associated With Tumoral Immunomodulation in Patients With Advanced Metastatic Melanoma
Human cancers express tumor antigens that can be targeted by cytolytic T lymphocytes (CTL).
 These antigens consist of a small peptide, derived from endogenous proteins, that is
 presented at the cancer cell's surface by an HLA class I molecule. Such antigenic peptides,
 including MAGE-3.A1 and NA17.A2, have been tested in experimental therapeutic vaccines to
 elicit CTL responses in cancer patients, mainly with metastatic melanoma. Up to now, only
 rare tumor responses have been observed.

 Tumor resistance to CTL killing is the most likely explanation for the poor effectiveness of
 cancer vaccines. This resistance is probably acquired by the tumor during its development and
 selected by its repetitive challenge with spontaneous anti-tumoral immune responses. The
 precise molecular mechanisms of tumor resistance remain unknown. The observation that
 tumor-infiltrating lymphocytes (TIL) purified from melanoma metastases can recognize and kill
 autologous tumor cells in vitro, whilst they seem unable to control tumor growth in vivo,
 suggests that this resistance is hosted by the tumor environment, rather than being the
 result of a generalized immune suppression.

 The investigators have developed a murine model of cutaneous graft rejection that mimics the
 situation in melanoma. Female CBA mice do not reject syngeneic male skin grafts, even though
 they mount a spontaneous CTL response against H-Y, a male specific minor histocompatibility
 antigen, following grafting. The investigators have tested various experimental procedures
 aimed at inducing effective graft rejection in these mice. This was obtained with a
 combination of IFN-α, IL-2, GM-CSF, each administered separately under the skin graft,
 associated with topical applications of imiquimod. All these agents are available as
 registered drugs. Based on this murine model of cutaneous allograft rejection, the
 investigators postulate that local immunomodulation with this combination can trigger an
 effective tumor rejection process, and induce a more efficient and long-lasting anti-tumoral
 immune response following peptide vaccination.
Patients will receive the following treatments:

 1. Vaccinations:

 The vaccine will be the MAGE-3.A1 and/or the NA17.A2 peptide, matching the patient's HLA
 type and the gene expression of his tumor. If both antigens are expressed, then the
 patient will receive both peptides.

 2. Local treatment with a combination of immunomodulatory drugs:

 This treatment will combine peritumoral injection of IL-2, IFN-α and GM-CSF (6000 IU, 100.000
 IU and 300 ng per tumor injected, respectively), as well as topical application of imiquimod
 (applied during 24h). The peritumoral injections of cytokines will be given on days
 +2,+3,+4,+7,+8 and +9, and the Aldara® cream will be applied on days +2 and +7 following
 vaccines 3 and 4. One or 2 cutaneous lesions will be treated, if there are 2 or more such
 lesions present at day 29 of the treatment, respectively.
Inclusion Criteria:

 -  1. Patients with histologically proven cutaneous melanoma at one of the following AJCC
 stages.Regional metastatic disease (any T; N2c or N3; M0). Distant metastatic disease
 (any T; any N; M1a, M1b or M1c*).*except uncontrolled brain metastasis.

 -  2. HLA-A1 or HLA-A2 (by serology or molecular biology).

 -  3. MAGE-3 gene expression by the tumor if patient is HLA-A1 and/or NA17 gene
 expression by the tumor if patient is HLA-A2 (determined by RT-PCR analysis).

 -  4. Measurable Disease (RECIST v1.1)Patients must have at least 2 cutaneous metastases,
 suitable for peri-tumoral injection and surgical resection, with their largest
 diameter equal to or greater than 5 mm.

 -  5. Age ≥ 18 years.

 -  6. Karnosky Performance status (KPS) ≥70 or WHO performance status of 0 or 1

 -  7. Expected survival of at least 6 months.

 -  8. Normal laboratory values : Platelet count ≥100x103/μL,Leucocyte count ≥ 3x103/μL,
 Hemoglobin ≥ 9 g/dL, ASAT and ALAT ≤ 2xUNL, Serum creatinine ≤1.5xUNL, Total bilirubin
 ≤ 1.5xUNL, LDH ≤ 1.5xUNL

 -  9. Viral serology : negative antibodies for HCV & HIV; negative antigens for HBV.

 -  10. Patient should agree to perform biopsies and blood collections for translational
 research.

 -  11. Signed informed consent from the patient or legal representative must be obtained.

 Exclusion Criteria:

 1. Uncontrolled brain or central nervous system metastases.

 2. Previous treatment for the melanoma within 6 weeks from inclusion, with any reagent
 known to modulate the immune system such as a cancer vaccine, interferon-alpha,
 interleukins or anti-CTLA-4 antibodies.

 3. Previous chemotherapy, radiotherapy, corticotherapy, or other immune suppressive
 therapy within 4 weeks from inclusion.

 4. Clinically significant cardiovascular disease (including cardiac insufficiency NYHA
 grade III and IV, unstable angina, arrythmia, myocardial infarction, symptomatic
 congestive heart failure) in the past 12 months before enrollment.

 5. Other serious acute or chronic illnesses, e.g. active infections requiring
 antibiotics, bleeding disorders or other conditions requiring concurrent medications
 not allowed during this study.

 6. Other malignancy within 3 years prior to entry in the study, except for treated
 non-melanoma skin cancer and in situ cervical carcinoma.

 7. Active immunodeficiency disease or autoimmune disease (Vitiligo is not an exclusion
 criterion).

 8. Lack of availability for immunological and clinical follow-up assessments.

 9. Participation in any other clinical trial involving another investigational agent
 within 4 weeks prior to enrollment.

 10. Subject pregnant or breastfeeding, or planning to become pregnant within 6 months
 after the end of treatment.

 11. Subject (male or female) not willing to use highly effective methods of contraception
 (per institutional standard) during treatment and for 6 months after the end of
 treatment.