Study Evaluating Impact of IL-7 on CD4 Lymphopenia, Risks of Severe Haematological Toxicity and Tumor Progression in Metastatic Breast Cancer Patients

• to determine if CYT107 treatment enables to reduce the incidence of severe haematological toxicity (any type of haematological toxicity Grade ≥ 3) post-chemotherapy [ Time Frame: at the end of study M12 ]
• To assess the impact of CYT107 on progression-free survival [ Time Frame: at the end of study (M12) ]
  Time from randomisation to first evidence of progression or death of any cause.
• To assess the impact of CYT107 on compliance to chemotherapy regimen (dose intensity, number of chemotherapy cycles). [ Time Frame: at the end of study (M12) ]
  Number of CT cycles, CT dose delays and/or reduction, CT discontinuation
• To assess the impact of CYT107 on CD4 lymphopenia over the study period [ Time Frame: at the end of study (M12) ]
  Evolution of CD4 count from Day 0 to end of study visit
• to evaluate if CYT107 treatment will selectively stimulate the proliferation and activation of peripheral immune subsets (analysis of phenotype and activation status of peripheral immune e sub-populations) [ Time Frame: D0, D21, D57, D78 and at end of study M12 ]
  Measure of frequency and activation status of circulating immune subpopulations on fresh whole blood. Multi-parametric marker sets (6-8 markers) will be used to analyse phenotype of immune subpopulations (TCD4+, TCD8+, Treg, T, NK, DC) and their activation status (PD1, ICOS, CD39, CD73, CD62L, CCR7, CD45RO, CD45RA, CD86).
• to evaluate if CYT107 treatment will selectively improve the functional response of T cells, DC subsets and NK cells [ Time Frame: D0, D21, D57, D78 and at the end of study M12 ]
  Analysis of the functional response of T cells, DC subsets and NK cells
• to evaluate if CYT107 treatment will is able to revert tolerogenic immune burden to increase specific anti-tumor response (measure of antigen specific CD8 response, measure of cytokine plasmatic levels) [ Time Frame: D0, D21, D57, D78 and at the end of study M12 ]
  • Analysis of tumor associated antigen (TAA) specific CD8 responses
  • Quantification of circulating cytokines including mainly, but not limited to, IL-6, IL-2, IFN, VEGF, TNF, IL-15,F FGF using Luminex technology and VEGF, TGF, IL-7R by Elisa.
• to evaluate if CYT107 treatment will enable to increase TCR diversity (analysis of combinatorial diversity). [ Time Frame: D0, D21, D57, D78 and at the end of study M12 ]
  Evaluation of T cell receptor diversity using ImmuneTraCkeR test and Constel'ID software (ImmunID Technologies, Grenoble, France).
• To assess the impact of CYT107 treatment on overall incidence of side effects [ Time Frame: after 12 weeks of treatment ]
  Number of patients with AEs (any type any grade) using NCI-CTCAE scale (version 4.0) from D0 to W12

• to determine if CYT107 treatment enables to reduce the incidence of severe haematological toxicity (any type of haematological toxicity Grade ≥ 3) post-chemotherapy [ Time Frame: at the end of study M12 ]
• To assess the impact of CYT107 on progression-free survival [ Time Frame: at the end of study (M12) ]
  Time from randomisation to first evidence of progression or death of any cause.
• To assess the impact of CYT107 on compliance to chemotherapy regimen (dose intensity, number of chemotherapy cycles). [ Time Frame: at the end of study (M12) ]
  Number of CT cycles, CT dose delays and/or reduction, CT discontinuation
• To assess the impact of CYT107 on CD4 lymphopenia over the study period [ Time Frame: at the end of study (M12) ]
  Evolution of CD4 count from Day 0 to end of study visit
• to evaluate if CYT107 treatment will selectively stimulate the proliferation and activation of peripheral immune subsets (analysis of phenotype and activation status of peripheral immune e sub-populations) [ Time Frame: D0, D21, D63, D184 and at end of study M12 ]
  Measure of frequency and activation status of circulating immune subpopulations on fresh whole blood. Multi-parametric marker sets (6-8 markers) will be used to analyse phenotype of immune subpopulations (TCD4+, TCD8+, Treg, T, NK, DC) and their activation status (PD1, ICOS, CD39, CD73, CD62L, CCR7, CD45RO, CD45RA, CD86).
• to evaluate if CYT107 treatment will selectively improve the functional response of T cells, DC subsets and NK cells [ Time Frame: D0, D21, D63, D184 and at the end of study M12 ]
  Analysis of the functional response of T cells, DC subsets and NK cells
• to evaluate if CYT107 treatment will is able to revert tolerogenic immune burden to increase specific anti-tumor response (measure of antigen specific CD8 response, measure of cytokine plasmatic levels) [ Time Frame: D0, D21, D63, D184 and at the end of study M12 ]
  • Analysis of tumor associated antigen (TAA) specific CD8 responses
  • Quantification of circulating cytokines including mainly, but not limited to, IL-6, IL-2, IFN, VEGF, TNF, IL-15,F FGF using Luminex technology and VEGF, TGF, IL-7R by Elisa.
• to evaluate if CYT107 treatment will enable to increase TCR diversity (analysis of combinatorial diversity). [ Time Frame: D0, D21, D63, D184 and at the end of study M12 ]
  Evaluation of T cell receptor diversity using ImmuneTraCkeR test and Constel'ID software (ImmunID Technologies, Grenoble, France).
• To assess the impact of CYT107 treatment on overall incidence of side effects [ Time Frame: after 12 weeks of treatment ]
  Number of patients with AEs (any type any grade) using NCI-CTCAE scale (version 4.0) from D0 to W12

The purpose of the study is to evaluate the impact of an immunotherapy by IL-7 on CD4 lymphopenia, risks of severe haematological toxicity and tumor progression in metastatic breast cancer patients.

The primary objective is to determine the optimal schedule to deliver CYT107 during chemotherapy based on restoration of CD4 count.

This study is a phase II, randomised, double-blind, placebo-controlled, single-centre.

24 patients will be included in the study.

A key secondary objective is to determine if CYT107 treatment enables to reduce the incidence of severe haematological toxicity (any type of haematological toxicity Grade ≥ 3) post-chemotherapy.

Other secondary objectives are to assess the impact of CYT107 treatment on the following parameters:

• Overall incidence of side effects (any type any grade)
• Progression-free survival (PFS)
• Compliance to chemotherapy regimen (dose intensity, number of chemotherapy cycles).
• CD4 lymphopenia over the study period

Exploratory biological markers

A series of biomarkers analyses will be performed to evaluate if CYT107 treatment will:

• selectively stimulate the proliferation and activation of peripheral immune subsets (analysis of phenotype and activation status of peripheral immune e sub-populations)
• selectively improve the functional response of T cells, DC subsets and NK cells.
• is able to revert tolerogenic immune burden to increase specific anti-tumor response (measure of antigen specific CD8 response, measure of cytokine plasmatic levels)
• enable to increase TCR diversity (analysis of combinatorial diversity).

• Drug: placebo
  Placebo before the 1st (D0, D7, D14)and during the 3rd CT cycle (D57, D64, D71)
• Drug: interleukin 7
  patients will receive an induction cycle of CYT107 (10µg/kg/week subcutaneously for 3 weeks) before the 1st CT cycle (D0, D7, D14) and the placebo during the 3rd CT cycle (D57, D64, D71)
• Drug: interleukin 7
  patients will receive the placebo before the 1st CT cycle (D0, D7, D14) and a delayed treatment with CYT107 (10µg/kg/week subcutaneously for 3 weeks) during the 3rd CT cycle (D57, D64, D71)
• Drug: interleukin 7
  patients will receive an induction cycle of CYT107 (10µg/kg/week subcutaneously for 3 weeks) before the 1st CT cycle (D0, D7, D14) and a maintenance cycle of IL-7 (10µg/kg/week subcutaneously for 3 weeks) during the 3rd CT cycle (D57, D64, D71)

• Placebo Comparator: Placebo Arm
  the patients will receive Placebo before the 1st and during the 3rd CT cycle (N=6)
  Intervention: Drug: placebo
• Experimental: CYT107 treatment before CT
  patients will receive an induction cycle of CYT107 (10µg/kg/week subcutaneously for 3 weeks) before the 1st CT cycle and the placebo during the 3rd CT cycle (N=6)
  Interventions:

  • Drug: placebo
  • Drug: interleukin 7
• Experimental: CYT107 treatment during CT
  patients will receive the placebo before the 1st CT cycle and a delayed treatment with CYT107 (10µg/kg/week subcutaneously for 3 weeks) during the 3rd CT cycle (N=6)
  Intervention: Drug: interleukin 7
• Experimental: CYT107 treatment before and during CT
  patients will receive an induction cycle of CYT107 (10µg/kg/week subcutaneously for 3 weeks) before the 1st CT cycle and a maintenance cycle of IL-7 (10µg/kg/week subcutaneously for 3 weeks) during the 3rd CT cycle (N=6).
  Intervention: Drug: interleukin 7

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Inclusion Criteria:

• Female aged more than 18 years
• Histologic diagnosis of metastatic breast cancer to be treated with capecitabine at study entry. NB: Patients previously treated with capecitabine are eligible only if more than 6 months have elapsed since the last capecitabine intake.
• Lymphopenic (i.e. with at least one value of lymphocyte count 1500/µL within 15 days before Day 0).
• Performance status ECOG of 0, 1,2 or 3
• Life expectancy ≥ 6months

• Adequate bone marrow, hepatic and renal function as follows:

  • Neutrophils ≥ 1,000/µL
  • Platelets ≥ 100 109/µL
  • ASAT, ALAT, or Alkaline Phosphatase ≤ 2.5 x ULN
  • Total Bilirubin ≤ 1.5 x ULN
  • INR ≤ 1.5
  • Calculated creatinin clearance ≥ 60mL/min (Cockcroft formula or MDRD formula for patients older than 65 years old)- Ability to understand and sign informed consent
• Covered by a medical insurance.

Exclusion Criteria:

• Prior history of other malignancies other than breast cancer (except for basal cell or squamous cell carcinoma of the skin or carcinoma in situ of the cervix) unless the subjects has been free of the disease for at least 3 years.
• No resolution of specific toxicities related to any prior anti-cancer therapy to Grade ≤2 according to the NCI CTCAE v.4.0 (except lymphopenia, alopecia and neuropathy)
• Wash out period of less than 5 times the half-life of previous anti-cancer treatment before study entry, except if previous chemotherapy treatment before study entry. NB: For patient previously treated by hormonotherapy, a wash out period of 1 week will be sufficient
• Uncontrolled hypertension (i.e., resting systolic blood pressure greater than140 mmHg or resting diastolic blood pressure greater than 90 mmHg), despite pharmacologic antihypertensive treatment, confirmed with a second blood pressure measurement done later in the same day
• History of lymphoid malignancy (e.g. Hodgkin disease, non Hodgkin lymphoma, Leukemia).
• History of splenectomy or hematologic disease associated with hypersplenism, such as gamma or beta-thalassemia, hereditary spherocytosis, Gaucher's disease, or autoimmune hemolytic anemia.
• Any cardiac, pulmonary, thyroid, renal, hepatic, neurological severe/uncontrolled concurrent medical disease that in the opinion of the investigator could cause unacceptable safety risks or compromise compliance with the protocol
• Any history of severe auto-immune disease
• Hepatitis B antigen (HBs Ag) positive, Hepatitis C (HCV Ab) antibody positive or HCV RNA detectable
• Documented HIV-1 positivity
• History of cardiovascular disorders grade >2 (NYHA) within 6 months preceding the inclusion
• Active uncontrolled viral, fungal or bacterial infection
• Active drug or alcohol use or dependence that, in the opinion of the investigator, would interfere with adherence to study requirements (participants must agree to refrain from substance abuse use during the entire course of the study)
• Pregnant or breast-feeding women
• No use of effective birth control methods for women of childbearing potential
• Any contraindications to capecitabine treatment (refer to Xeloda SPC Appendix 11) and to any other anti-cancer treatment authorized as per protocol (refer to respective SPC for specific contraindications)

• Ministry of Health, France
• Cytheris, Inc.