Lymphoma with secondary central nervous system (CNS) involvement represents one of the most aggressive malignancies, with poor prognosis and high mortality. New diagnostic tools for its early detection, response evaluation, and CNS relapse prediction are needed. We analyzed circulating microRNAs in the cerebrospinal fluid (CSF) and plasma of 162 patients with aggressive B-cell non-Hodgkin's lymphomas (B-NHL) and compared their levels in CNS-involving lymphomas versus in systemic lymphomas, at diagnosis and during treatment and CNS relapse. We identified a set of five oncogenic microRNAs (miR-19a, miR-20a, miR-21, miR-92a, and miR-155) in CSF that detect, with high sensitivity, secondary CNS lymphoma involvement in aggressive B-NHL, including DLBCL, MCL, and Burkitt lymphoma. Their combination into an oncomiR index enables the separation of CNS lymphomas from systemic lymphomas or nonmalignant controls with high sensitivity and specificity, and high Receiver Operating Characteristics (DLBCL AUC = 0.96, MCL = 0.93, BL = 1.0). Longitudinal analysis showed that oncomiR levels reflect treatment efficacy and clinical outcomes, allowing their monitoring and prediction. In contrast to conventional methods, CSF oncomiRs enable detection of early and residual CNS involvement, as well as parenchymal involvement. These circulating oncomiRs increase 1-4 months before CNS relapse, allowing its early detection and improving the prediction of CNS relapse risk in DLBCL. Similar effects were detectable, to a lesser extent, in plasma.

1st Department of Medicine Charles University General Hospital 128 08 Prague Czech Republic

Biocev 1st Faculty of Medicine Charles University 252 50 Vestec Czech Republic

Department of Cell Therapy Institute of Haematology and Blood Transfusion 128 20 Prague Czech Republic

Department of Haematology University Hospital Kralovske Vinohrady and 3rd Faculty of Medicine Charles University 100 34 Prague Czech Republic

Faculty of Mathematics and Physics Charles University 186 75 Prague Czech Republic

Institute of Pathological Physiology 1st Faculty of Medicine Charles University 128 53 Prague Czech Republic

Section of Experimental Haematology Leeds Institute of Cancer and Pathology St James's University Hospital University of Leeds Leeds LS2 9JT UK

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Outcome of patients with diffuse large B-cell lymphoma and testicular involvement - real world data