NOVEL EPIGENETIC TEST DISTINGUISHES MDS, AML AND CONTROLS THROUGH PERIPHERAL BLOOD ANALYSIS
Miriam Neaman, Department of Chemistry, Raymond and Beverly Sackler Faculty of Exact Sciences, Tel Aviv University, Department of Hematology, Tel Aviv Sourasky Medical Center. Matan Fainzilber, Department of Statistics and Operations Research, Tel Aviv University.
Jasline Deek, Department of Chemistry, Raymond and Beverly Sackler Faculty of Exact Sciences, Tel Aviv University. Rita Haldar, Department of Chemistry, Raymond and Beverly Sackler Faculty of Exact Sciences, Tel Aviv University. Amit Manor, Gray Faculty of Medical & Health Sciences, Tel Aviv University. Mika Bell, JaxBio Technologies Ltd. Yakir Moshe, Department of Hematology, Tel Aviv Sourasky Medical Center. Irit Mazza Avivi, Department of Hematology, Tel Aviv Sourasky Medical Center, Gray Faculty of Medical & Health Sciences, Tel Aviv University. Moshe Mittelman, Department of Hematology, Tel Aviv Sourasky Medical Center, Gray Faculty of Medical & Health Sciences, Tel Aviv University. Karolis Sablauskas, Institute of Data Science and Digital Technologies, Vilnius University, Vilnius, Lithuania; Hematology, Oncology and Transfusion Medicine Center, Vilnius University Hospital Santaros Klinikos, Vilnius, Lithuania. Efstathios Kastritis, Department of Clinical Therapeutics, National and Kapodistrian University of Athens, School of Medicine, Athens, Greece. Tomas Papajik, Department of Hemato-Oncology, Faculty of Medicine and Dentistry, Palacky University and University Hospital, Olomouc, Czech Republic. Eva Kriegova, Department of Immunology, Faculty of Medicine and Dentistry, Palacky University and University Hospital, Olomouc, Czech Republic. Amit Moscovich, Department of Statistics and Operations Research, Tel Aviv University. Yuval Ebenstein, Department of Chemistry, Raymond and Beverly Sackler Faculty of Exact Sciences, Tel Aviv University, Department of Biomedical Engineering, Faculty of Engineering, Tel Aviv Univesity.​
Objectives: Aberrant DNA methylation patterns are well-documented in acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS), yet epigenetic markers remain absent from diagnostic or classification scores. We combined direct chemoenzymatic 5mC and 5hmC labeling with microarray technology for enhanced epigenetic profiling of AML and MDS. We aim to introduce a simple non-invasive tool for AML and MDS detection and monitoring using epigenetic markers.
Methods: Peripheral blood samples from 53 AML, 36 MDS patients, and 50 controls were collected across four European centers (NCT05735704). We applied direct chemoenzymatic 5hmC and 5mC labeling developed by our lab, producing dual-labeled DNA with distinct fluorophores. Labeled DNA was hybridized to custom microarrays targeting 13,000 genomic loci relevant to AML methylation. Fluorophore intensities representing 5hmC and 5mC levels were analyzed using Welch's t-statistics to identify differentially methylated loci distinguishing between cohorts as biomarkers. Performance was evaluated using leave-one-out cross-validation with logistic regression for individual markers and multichannel integration.
Results: Preliminary results demonstrated successful discrimination between AML, MDS, and controls using 500 differential markers from 13,000 loci. Multichannel analysis combining both markers achieved superior performance versus single-marker approaches. For AML versus control, 5hmC and 5mC analyses yielded 94.9% and 97.2% AUC, respectively, while multichannel integration achieved 98.4% AUC with 94% sensitivity and 90% specificity. MDS detection improved with combined analysis: 97% AUC, 94% sensitivity, and 98% specificity. Multichannel analysis distinguished MDS from AML with 98.5% AUC, correctly classifying 98% of AML and 92% of MDS samples.
Conclusions: AML and MDS can be accurately detected from peripheral blood using our novel method with combined 5hmC and 5mC analysis. This assay potentially assists physicians in clinical decision-making prior to performing invasive and costly bone marrow testing. This first-in-class multichannel epigenetic analysis demonstrates significant clinical utility for early MDS and AML detection and monitoring MDS progression to AML.