Aberrant DNA methylation at specific genetic loci is a key molecular feature of juvenile myelomonocytic leukemia (JMML) with poor prognosis. Using quantitative high-resolution mass spectrometry, we identified RASA4 isoform 2, which maps to chromosome 7 and encodes a member of the GAP1 family of GTPase-activating proteins for small G proteins, as a recurrent target of isoform-specific DNA hypermethylation in JMML (51% of 125 patients analyzed). RASA4 isoform 2 promoter methylation correlated with clinical parameters predicting poor prognosis (older age, elevated fetal hemoglobin), with higher risk of relapse after hematopoietic stem cell transplantation, and with PTPN11 mutation. The level of isoform 2 methylation increased in relapsed cases after transplantation. Interestingly, most JMML cases with monosomy 7 exhibited hypermethylation on the remaining RASA4 allele. The results corroborate the significance of epigenetic modifications in the phenotype of aggressive JMML.

Department of Pediatric Hematology Oncology; IRCCS Ospedale Bambino Gesu; Rome Italy; University of Pavia; Pavia Italy

Department of Pediatric Oncology Hematology; Erasmus Medical Center; Rotterdam The Netherlands; Dutch Children's Oncology Group; The Hague Netherlands

Department of Pediatrics; Aarhus University Hospital Skejby; Aarhus Denmark

Dept of Pediatric Hematology and Oncology; 2nd Faculty of Medicine; Charles University and University Hospital Motol Praha; Czech Pediatric Hematology Working Group ; Prague Czech Republic

Dept of Pediatric Hematology Oncology; Ghent University Hospital; Ghent Belgium

Division of Biostatistics; German Cancer Research Center; Heidelberg Germany

Division of Epigenomics and Cancer Risk Factors ; German Cancer Research Center; Heidelberg Germany

Division of Epigenomics and Cancer Risk Factors ; German Cancer Research Center; Heidelberg Germany; The German Cancer Consortium; Heidelberg Germany

Division of Hematology; University Children's Hospital; Zurich Switzerland

Division of Pediatric Hematology Oncology; University Medical Center; Freiburg Germany

Pediatric Hematology Oncology; Fondazione IRCCS;Policlinico San Matteo; Pavia Italy

St Anna Children's Hospital and Children's Cancer Research Institute; Department of Pediatrics; Medical University of Vienna; Vienna Austria

The German Cancer Consortium; Heidelberg Germany; Division of Pediatric Hematology Oncology; University Medical Center; Freiburg Germany

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