BMS_PD-L1_onco : Assessment of the PD-L1 Protein as a Biomarker in Oncology and Hematology

Study Overview
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Brief Summary
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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).

Detailed Description
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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

Official Title
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BMS_PD-L1_onco : Assessment of the PD-L1 Protein as a Biomarker in Oncology and Hematology

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

Participation Criteria
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Eligibility Criteria
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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 < 18 years et > 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

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 75 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| DLBCL (diffuse large B-cell lymphoma)<br>DLBCL (diffuse large B-cell lymphoma) | |
| Hodgkin's lymphoma<br>Hodgkin's lymphoma | |
| metastatic breast cancer<br>metastatic breast cancer or with lymph node involvement | |
| immune thrombocytopenia (ITP)<br>immune thrombocytopenia (ITP) | |
| healthy volunteers<br>healthy volunteers | |
| non-small cell lung cancer<br>non-small cell lung cancer | |

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Description of one or several blood cell types producing soluble PD-L1 in DLBCL, metastatic breast cancer, Hodgkin's lymphoma and non-small cell lung cancer | Description of one or several blood cell types producing soluble PD-L1 in DLBCL, metastatic breast cancer, Hodgkin's lymphoma and non-small cell lung cancer | 4 years | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Analysis of PD-L1 membrane protein expression on circulating tumor cells by multiparameter flow cytometry and Veridex® in DLBCL and metastatic breast cancer, and bone marrow tumor cells by flow cytometry in DLBCL | Analysis of PD-L1 membrane protein expression on circulating tumor cells by multiparameter flow cytometry and Veridex® in DLBCL and metastatic breast cancer, and bone marrow tumor cells by flow cytometry in DLBCL | 4 years | 
| Analysis of PD-L1 membrane protein expression on circulating endothelial cells with the Veridex® technology in DLBCL, Hodgkin's lymphoma and metastatic breast cancer (subpart ended in late 2012) | Analysis of PD-L1 membrane protein expression on circulating endothelial cells with the Veridex® technology in DLBCL, Hodgkin's lymphoma and metastatic breast cancer (subpart ended in late 2012) | 4 years | 
| Analysis of PD-L1 membrane protein expression on monocytes, B and T lymphocytes in all cohorts | Analysis of PD-L1 membrane protein expression on monocytes, B and T lymphocytes in all cohorts | 4 years | 
| Development of an ELISPOT technique to detect soluble PD-L1 in the supernatants of sorted primary cells (subpart ended mid 2013) | Development of an ELISPOT technique to detect soluble PD-L1 in the supernatants of sorted primary cells (subpart ended mid 2013) | 4 years | 
| Evaluation of the techniques (by immunomagnetic or cell-sorting) used to separate circulating tumor cells expressing PD-L1 | Evaluation of the techniques (by immunomagnetic or cell-sorting) used to separate circulating tumor cells expressing PD-L1 | 4 years | 
| Correlation between the PD-L1 expression *) in the plasma, **) in the tumor and ***) in the bronchoalveolar liquid in non-small cell lung cancer | Correlation between the PD-L1 expression *) in the plasma, **) in the tumor and ***) in the bronchoalveolar liquid in non-small cell lung cancer | 4 years | 
| Evaluation of the response to treatment according to soluble PD-L1 expression in non-small cell lung cancer | Evaluation of the response to treatment according to soluble PD-L1 expression in non-small cell lung cancer | 4 years | 
| Evaluation of the susceptibility to develop a disease according to PD-L1 gene SNP in DLBCL and non-small cell lung cancer | Evaluation of the susceptibility to develop a disease according to PD-L1 gene SNP in DLBCL and non-small cell lung cancer | 4 years | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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diffuse large B-cell lymphoma (DLBCL), Hodgkin lymphoma, metastatic breast cancer., non-small cell lung cancer