bms_pd-l1_onco : assessment of the pd-l1 protein as a biomarker in oncology and hematology

Conditions: Diffuse Large B-cell Lymphoma; Hodgkin Lymphoma; Metastatic Breastcancer; Non-small Cell Lung Cancer

Summary: diffuse large b-cell lymphomas (dlbcls) represent 25 to 30% of adult non-hodgkin lymphomas in western countries. dlbcls are aggressive cancer but potentially curable with multi-agent chemotherapy. whereas r-chop regimen has led to a marked improvement in survival, this disease remains a biologically heterogeneous entity. new therapeutic strategies are required including identification of patients' subgroups with different prognostic. this project is based on bms_lytrans and goelams 075 clinical trial. a study of whole blood transcriptome in 75 dlbcl patients and in 87 controls showed that pd-l1 (cd274) gene was overexpressed in dlbcl patients. preliminary results demonstrated that pd-l1 is detected in plasma of dlbcl patients with a significantly higher concentration than in controls. this protein was selected as a potential biomarker because of its established role in anti-tumoral immunity. interaction between pd-l1 and its receptor pd-1 is known to inhibit activation of immune responses by inducing t-lymphocytes anergy and/or apoptosis. moreover, a direct involvement of pd-l1 in the protection of cancer cells from lysis by activated t lymphocytes has been demonstrated. pd-l1 expression has been described in several solid tumours, including ovary cancer, breast cancer, colon cancer, renal cell carcinoma, non-small cell lung carcinoma and in hematological malignancies such as t-nhl, mm and hodgkin's lymphoma. furthermore the expression of pd-l1 by tumour cells is associated with poor prognosis. the blockade of pd-l1/pd-1 axis may represent a novel therapeutic approach in aggressive cancers. these first results incite to identify the cells releasing soluble pd-l1 and to investigate its role in the anti-tumoral immunity in dlbcl patients. the aim of this study is to identify cells producing soluble pd-l1 in dlbcl patients at diagnosis in comparison to others tumours known to express pd-l1 (metastatic breast cancer, hodgkin's lymphoma, non-small cell lung cancer).

secondary purposes are : * to confirm the presence of plasma soluble form of pd-l1 in others malignancies * to study surface expression of pd-l1 on circulating tumour cells with multiparameter fow cytometry and veridex® technology in dlbcl and metastatic breast cancer patients * to study surface expression of pd-l1 on circulating endothelial cells in dlbcl, hodgkin lymphoma and metastatic breast cancer patients (subpart ended in late 2012) * to study surface expression of pd-l1 on different types of leukocytes (monocytes, b and t lymphocytes) * to separate circulating tumour cells expressing pd-l1 by immunomagnetic or cell-sorting method * to develop elispot technique to study the release of soluble pd-l1 in culture supernatants of selected cells (subpart ended mid 2013) * to evaluate the correlation between the expression of pd-l1 in the plasma and \*) the expression of pd-l1 in the tumor, \*\*) the expression of pd-l1 and other molecules in the bronchoalveolar liquid (whenever available from routine) in non-small cell lung cancer * to evaluate the response to treatment according to plasma pd-l1 expression in non-small cell lung cancer * to evaluate the susceptibility to develop a disease according to the single nucleotide polymorphisms of the pd-l1 gene in dlbcl and non-small cell lung cancer * constitution of the different cohorts and collection of samples main cohort : de novo dlbcl at diagnosis secondary cohorts: hodgkin's lymphoma, metastatic breast cancer, non small cell lung cancer control cohorts : healthy volunteers (blood donors), patients with immune thrombocytopenia (itp) * quantification of plasma soluble pd-l1 in the different cohorts

Eligibility criteria:
inclusion criteria: general inclusion criteria : * age ≥ 18 years and ≤ 75 years, * life expectancy more than 4 months * signed informed consent obtained * social security affiliation is mandatory * non previously treated (even by corticotherapy), * hiv negative, hbs negative, hcv negative inclusion criteria for dlbcl patients : * a biopsy proven diagnosis of de novo dlbcl according to the current who criteria, * immunohistochemistry for gcb/nongc classification according to hans' algorithm * patients with advanced-stage disease defined as ann arbor stages iii or iv, or stages i or ii with bulky disease (\>7cm) inclusion criteria for non-small cell lung cancer patients : * a biopsy proven diagnosis of de novo non-small cell lung cancer (all stages) according to the current who criteria inclusion criteria for hodgkin's lymphoma patients : * a biopsy proven diagnosis of hodgkin's lymphoma according to the current who criteria inclusion criteria for metastatic breast cancer or with lymph node involvement : * a biopsy proven diagnosis infiltrating lobular or ductal breast carcinoma * with lymph node involvement or metastasis inclusion criteria for patients with immune thrombocytopenia (itp) : * primary itp was defined by the iwg as a platelet count less than 100 g/l in the absence of other causes or disorders that may be associated with thrombocytopenia. * bone marrow examination excluding a central aetiology of thrombocytopenia inclusion criteria for healthy volunteers : - inclusion criteria for blood donation according to the etablissement français du sang (efs) criteria exclusion criteria: general non-inclusion criteria : * age \ 75 years, * pregnant women, * person legally involved in a case * no social security affiliation * signed informed consent not obtained, * preliminary treatment (even corticoid treatment). * hiv positive, hbs positive, hcv positive non-inclusion criteria for dlbcl patients : * lymphoma other than dlbcl, * transformation of a low grade lymphoma to a high grade lymphoma (dlbcl), * extranodal marginal zone lymphoma of malt lymphoma, * post-transplant lymphoproliferative disorders, * lymphoblastic lymphoma, * burkitt's lymphoma, * carcinoma or history of carcinoma except in situ cervical carcinoma. non-inclusion criteria for non-small cell lung cancer patients : none non-inclusion criteria for hodgkin patients : - non hodgkin's lymphoma non-inclusion criteria for metastatic breast cancer or with lymph node involvement : * carcinoma other than infiltrating lobular or ductal breast carcinoma * chemotherapy in 30 days preceding the inclusion * hormonotherapy in 7 days preceding the inclusion * carcinoma or history of carcinoma except in situ cervical carcinoma. * hemoglobin level \< 10g/dl non-inclusion criteria for patients with immune thrombocytopenia (itp) : - central aetiology of the thrombocytopenia non-inclusion criteria for healthy volunteers : - exclusion criteria for blood donation according to the etablissement français du sang (efs) criteria

Keywords: diffuse large b-cell lymphoma (dlbcl) hodgkin lymphoma metastatic breast cancer. non-small cell lung cancer