DNA methylation profiling of sorted cells from myelofibrosis patients reveals aberrant epigenetic regulation of immune pathways and identifies early MPN driver genes

Helene Myrtue Nielsen, Christen Lykkegaard Andersen, Lasse Sommer Kristensen, Fazila Asmar, Torben A Kruse, Mads Thomassen, Thomas Stauffer Larsen, Vibe Skov, Erik Sørensen, Henrik Ullum, Caroline Riley, Lise Lotte Hansen, Ole Weis Bjerrum, Hans Carl Hasselbalch, Vasu Punj, K Grønbæk

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Abstract

Background: Primary myelofibrosis (PMF) belongs to the heterogeneous group of chronic myeloproliferative neoplasms (MPN) together with essential thrombocytosis (ET) and polycythemia vera (PV). It has been suggested that these neoplasms represent a biological continuum from early cancer stage (ET, PV) toadvancedMF. Multiple studies report frequent mutations in epigenetic regulators. However, the association to epigenetic changes and the role of epigenetic aberrations in different cell populations is still unknown. Aims: We therefore performed DNA methylation profiling of sorted cells from MF patients to unravel pathways contributing to disease phenotype and gain insight into MF pathogenesis. As an aberrant DNA methylation pattern may be an early event in tumorigenesis and may be crucial for progression of the malignant clone towards the more aggressive forms of MPN, we further aimed to identify candidate driver genes. Methods: Peripheral blood samples from 16 MF patients and BM (bone marrow) and peripheral blood from 3 healthy age matched controls were sorted in CD34+ cells, granulocytes and mononuclear cells, and analysed for differential methylated regions using Illumina Infinium HumanMethylation 450K BeadChip. Candidate genes were validated by pyrosequencing in a second cohort of 30 MF patients where DNA was extracted from full blood (PB). To identify potential driver genes, the DNA methylation status of candidate genes was likewise analyzed in PB from a larger cohort consisting of 60 ET and PV patients. Results: The number of differentially methylated CpG sites between MF cells and the respective counterparts from healthy donors differed extensively among the three cell populations analyzed. In MF CD34+ cells 1628 CpG sites were differentially methylated compared to normal CD34+ cells, and 519 and 213 differential methylated CpG sites were observed in MF granulocytes and MF mononuclear cells, respectively (DELTAbeta was set to 0.2 with an adjusted p-value
Original languageEnglish
Article numberP302
JournalHaematologica
Volume100
Issue numberS1
Pages (from-to)97
Number of pages1
ISSN0390-6078
Publication statusPublished - 2015
Event20th Congress of the European Hematology Association - Vienna, Austria
Duration: 11. Jun 201514. Jun 2015

Conference

Conference20th Congress of the European Hematology Association
Country/TerritoryAustria
CityVienna
Period11/06/201514/06/2015

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