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Is Trogocytosis a Predictive Marker of CAR-T Cell Response in Diffuse Large B-cell Lymphoma?

Not Applicable
Recruiting
Conditions
Diffuse Large B Cell Lymphoma
Interventions
Other: Flow cytometry analysis to determine the level of trogocytosis by effector immune cells in patients
Other: Flow cytometry analysis to determine the level of trogocytosis by effector immune cells in volunteers
Registration Number
NCT06352242
Lead Sponsor
University Hospital, Montpellier
Brief Summary

CAR-T cell therapy has improved survival in patients with relapsed or refractory diffuse large B-cell lymphoma (DLBCL R/R). However, only 65% of patients achieve a complete metabolic response after this treatment. To date, there is no predictive test for therapeutic response after injection of CAR-T cells. Recent studies have shown that the level of trogocytosis by immune cells correlates with the persistence of tumor cells in patients with hematological malignancies. Our main objective is to identify a phenotypic "signature" of trogocytosis predictive of therapeutic response 6 months after injection of CAR-T cells for DLBCL.

Detailed Description

The therapeutic use of CAR-T cells (Chimeric Antigen Receptor T-cells) has significantly improved the survival of patients with relapsed or refractory (R/R) diffuse large B-cell lymphoma (DLBCL). However, only 65% of patients achieve metabolic complete response after this treatment, and one year after CAR-T cells infusion, between 50 and 60% of patients have relapsed or died. Injection of CAR-T cells can also be responsible for serious immunologic and hematologic adverse events. To date, there is no predictive test for the therapeutic response or toxicity following injection of CAR-T cells.

Trogocytosis is a physiological mechanism by which an effector immune cell integrates fragments of the membrane of target cells into its membrane. These aberrant membrane markers can directly modify the functions of the cell that has acquired them. Although the physiological role of trogocytosis remains debated, recent studies have shown that the level of trogocytosis in immune effector cells is correlated with persistent tumor cells in patients with hematological malignancies.

Our main hypothesis is that, in DLBCL, the level of early trogocytosis, assessed by the aberrant expression of tumor markers on the surface of CAR-T cells and other immune effector cells between D0 and D30 after CAR-T cells infusion, correlates with therapeutic response at M6 and/or the occurrence of immunological or severe hematological CAR-T cells side-effects.

Recruitment & Eligibility

Status
RECRUITING
Sex
All
Target Recruitment
85
Inclusion Criteria
  • For patients

    • Patient who has given free and informed consent in writing for inclusion in the non-interventional CART-BANK protocol, and orally for the CARTROG protocol,
    • Patients over 18 years of age at the time of inclusion,
    • Diagnosis of LDGCB,
    • Decision to treat with anti-CD19 CAR-T cells,
    • Patient affiliated to or benefiting from a social security scheme.
  • For healthy volunteers:

    • Given free and informed oral consent for inclusion in the CARTROG protocol,
    • Donor between 18 and 70 years of age at the time of inclusion,
    • No history of solid cancer or hematological malignancy,
    • No known chronic pathology (e.g. hypertension, diabetes, etc.) and no daily treatment,
    • No surgical treatment within the last 6 months.
Exclusion Criteria
  • Patients who do not meet all the inclusion criteria,
  • Pregnant or breast-feeding patient,
  • Patient unable to follow the procedures and/or frequency of visits planned in the trial, for psychological, family, social or geographical reasons,
  • Patient unable to consent freely to inclusion, under guardianship, curatorship or safeguard of justice.

Study & Design

Study Type
INTERVENTIONAL
Study Design
PARALLEL
Arm && Interventions
GroupInterventionDescription
PatientsFlow cytometry analysis to determine the level of trogocytosis by effector immune cells in patientsBlood sample for Flow cytometry analysis
healthy volunteer donorFlow cytometry analysis to determine the level of trogocytosis by effector immune cells in volunteersBlood sample for Flow cytometry analysis
Primary Outcome Measures
NameTimeMethod
Identification of a phenotypic "signature" of trogocytosis predictive of failure to achieve a complete metabolic response for patients with diffuse large B-cell lymphoma.During 6 months after CAR-T cells injection

Using flow cytometry to determine the level of trogocytosis, the phenotypic "signature" of trogocytosis will be assessed by the AUC : area under the ROC (Receiver operating characteristic) curve; established on circulating CAR-T cells, T lymphocytes and NK cells of patients, and defined as: the percentage of cells aberrantly expressing different tumor antigens normally expressed on lymphoma cells (CD19, CD20, etc.) at the different analysis times and/or the median florescence intensity (MFI) of the expression of these markers at the different analysis times and/or the evolution of these percentages and the MFI between 2 analysis times.

Failure to obtain a complete metabolic response will be defined by: the absence of a complete metabolic response on the PET scans of D30, D90 and M6 in the absence of implementation of a new therapeutic line; or by the absence of complete metabolic response on all PET scans carried out before the implementation of a new therapeutic line.

Secondary Outcome Measures
NameTimeMethod
Identification of a phenotypic "signature" of trogocytosis predictive of the occurrence of grade II or more immunological adverse eventsDuring 60 days after CAR-T cells injection

Using flow cytometry analysis, the phenotypic "signature" will be assessed by the AUC used to predict the occurrence of immunological side effects (Cytokine Release Syndrome and immune effector cell-associated neurotoxicity syndrome) of grade II or + according to the American Society for Transplantation and Cellular Therapy ASTCT2 criteria. It will be established on circulating CAR-T, T lymphocytes and NK cells of patients, and defined as : the percentage of cells aberrantly expressing different tumor antigens normally expressed on lymphoma cells at the different analysis times,and/or the MFI of the expression of these markers at the different analysis times and/or the evolution of these percentages and the MFI between 2 analysis times.The occurrence of immunological side effects between D0 and D60 will be assessed by clinical and, if necessary, paraclinical examinations (lumbar puncture, brain MRI, etc.). The grade will be evaluated according to the ASTCT2 criteria.

Determination of the trogocytosis level of normal lymphocytes and NK cells in healthy subjectsAt enrollment

Flow cytometry analysis will be performed on each sample on the day of collection (at enrollment), to determine the level of trogocytosis which will be evaluated by simple analysis of the trogocytosis phenotype of normal lymphocytes and NK cells in healthy subjects. This result will be considered as a negative control to determine the phenotypic "signature" of trogocytosis of the primary and secondary outcomes above.

Identification of a phenotypic "signature" of trogocytosis predictive of the occurrence of serious hematological side effects.Between Day 30 and 6 months after CAR-T cells injection

Flow cytometry analysis will be performed to determine the level of trogocytosis by effector cells. The phenotypic "signature" of trogocytosis will be assessed by the AUC used to predict the occurrence of serious hematological side effects (grade III or + cytopenias according to Common Terminology Criteria for Adverse Events CTCAE 5.03 between D30 and M6 after injection of anti-CD19 CAR-T cells .It will be established on circulating CAR-T, T lymphocytes and NK cells of patients, and defined as :the percentage of cells aberrantly expressing different tumor antigens normally expressed on lymphoma cells at the different analysis times, and/or the MFI of the expression of these markers at the different analysis times and/or the evolution of these percentages and the MFI between 2 analysis times. The occurrence of hematological side effects between D30 and M6 will be assessed by blood counts. The grade will be evaluated according to the CTCAE v5.03 criteria.

Trial Locations

Locations (2)

Clinical hematology department, University Hospital

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Montpellier, France

Clinical Investigation Center, University Hospital

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Montpellier, France

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