MIF : a New Target to Eradicate the Pre-leukemic Clone in Acute Myeloid Leukemia
试验速览
- 阶段
- 不适用
- 状态
- 进行中（未招募）
- 入组人数
- 186
- 试验地点
- 1
- 主要终点
- MIF concentration in the blood and the bone marrow and MIF RNA expression
研究概览
简要总结
This study is an observational study of MIF involvement in retrospectively and prospectively included adult acute myeloid leukemia (AML). Standard care samples collected at diagnosis, after one course of treatment, at time of remission controls, and at time of relapse will be used.
The first objective is to determine which AMLs have pre-leukemic stem cells that overexpress MIF. Cytogenetic and molecular (NGS) profiling will be performed at diagnosis. Blood and bone marrow plasma, as well as bone marrow mononuclear cells will be collected and stored. The expression of MIF and its receptor (CD74 and CXCR4) will be analysed. Their prognostic value will be also tested.
The second objective is to test whether patients in complete remission have persistent pre-leukemic stem cells that overexpress MIF. Blood and bone marrow plasma, bone marrow mononuclear cells from patients in complete remission will be collected. MIF, CD74, and CXCR4 expression by hematopoietic cells at time of diagnosis and remission will be compared to determine which patients have a persistent overexpression/secretion of MIF. In the meantime, the persistence of initiating lesions in complete remission samples will be tested by NGS, digital PCR, FISH, or RT-PCR methods.
The third objective is to develop a pre-clinical model to target MIF in immuno-compromised mice (NSG mice) transplanted with primary AML cells and cells with pre-leukemic lesions. TET2 depletion leads to MIF over-expression/secretion by hematopoietic cells and improved multi-lineage NSG-repopulation capacity. MIF inhibitors and anti-MIF antibodies will be tested in these pre-clinical TET2-depleted models. Xenotransplantation of selected primary AML samples and xenotransplantation of TET2 depleted hematopoietic stem cells into NSG mice will be used.
The fourth objective is to understand how MIF is deregulated in pre-leukemic stem cells and how the MIF-dependent crosstalk between mesenchymal stromal cells (MSCs) and pre-leukemic stem cells or normal hematopoietic cells works.
The molecular mechanisms of MIF overexpression will be analyzed in hematopoietic stem and progenitor cells from normal and leukemic bone marrow, with a focus on cells depleted in TET2 or DNMT3A. To study the cross-talk between hematopoietic stem and progenitor cells, pre-leukemic stem cells, and bone marrow MSCs, co-culture experiments will be performed using available MSC cell lines and primary MSCs from healthy donors.
详细描述
The study aims at studying the involvement of MIF in adult AML.
- DESCRIPTION OF THE ELEMENT OR ELEMENTS UNDER INVESTIGATION 1.1. Measurement of MIF concentrations Plasma and BM supernatant from patients and controls (100 age-matched controls for blood and BM plasma, available at Tours Hospital, ClinicalTrials.gov #NCT02789839 ) will be analyzed by ELISA. BM supernatants will be obtained after a 2-step centrifugation procedure: 1 mL of fresh BM from AML patients or control BM will be centrifuged at 540 g for 5 min to recover viable cells in the pellet (for cytometric and expression studies); the supernatant will be centrifuged at 1000 g/15 min/4°C to prepare the plasma. MIF concentrations in BM and blood plasma will be systematically compared. As MIF concentrations may be affected by different factors, including inflammation, leukocytosis, and monocytosis, blood cell counts and plasma C-reactive protein levels will be measured and taken into account in the interpretation of the data.
1.2. Measurement of MIF, CD74, and CXCR4 expression Hematopoietic stem cells, progenitor cells and blasts from AML patients will be sorted according to the expression of CD34, CD38, CD90, CD45-RA, IL-3R antigens, taking into account the initial immunophenotype of the disease. Hematopoietic stem cells and progenitor cells from control BM samples will be sorted similarly. MIF, CD74 and CXCR4 RNA expression will be studied by quantitative RT-PCR in sorted cell subpopulations. The surface expression of CXCR4 and anti-CD74 as well as intracellular CD74 will be studied by multiparametric flow with cytometry.
1.3. Analysis of MIF, CD74, CXCR4 expression, and MIF concentrations at time of complete remission.
The analyses performed in a) and b) will be repeated in complete remission samples in 30 to 50 selected patients. Patients will be selected according to their response to therapy (assessment of complete remission and normal blood counts), and to the genotype of their disease (15-25 patients with either TET2 or DNMT3A mutations at diagnosis vs 15-25 patients without these mutations). Samples will be collected after two chemotherapy courses at time of remission evaluation. Whenever possible, bone marrow controls of patients in long term remission (> 1 year post diagnosis) will be collected and studied. In the meantime, NGS, ddPCR, FISH or RT-PCR techniques will be used to assess the persistence or absence of the initial AML lesions.
研究设计
- 研究类型
- Observational
- 观察模型
- Cohort
- 时间视角
- Other
入排标准
- 年龄范围
- 18 Years 至 —（Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 否
入选标准
- •Patient newly diagnosed with AML in Saint Antoine hospital or Tours University hospital
- •Age > or equal 18 years old,
- •Written informed consent signed
排除标准
- •Patient without social insurance
结局指标
主要结局
MIF concentration in the blood and the bone marrow and MIF RNA expression
时间窗: At Day 0
MIF concentration in the blood and the bone marrow (measured by ELISA), and MIF RNA expression assessed by quantitative RT-PCR in unsorted and sorted cell populations in order to determine which AMLs have pre-leukemic stem cells that overexpress MIF
次要结局
- Overall and leukemia free survivals(patient complete remission until the last patient visit, for 36 months period)
- The number of patients in complete remission with persistent pre-leukemic stem cells that over express MIF(at Day 0 and at time of complete remission or relapse until the last patient visit for a 36 months period.)
- Rate of MIF inhibitors and anti-MIF antibodies will be tested in preclinical models(at Day 0 and at time of complete remission or relapse until the last patient visit for a 36 months period.)
- The molecular mechanisms of MIF overexpression(at Day 0 and at time of complete remission or relapse until the last patient visit for a 36 months period.)