A Pyrosequencing Assay for the Detection of EZH2Y641 Mutations in Diffuse Large B cell Lymphoma and Follicular Lymphoma

Aims: Enhancer of Zeste Homolog 2 (EZH2) is a critical enzymatic subunit of Polycomb Repressive Complex 2 (PRC2), which provides inhibitory regulation of gene transcription through trimethylation of histone H3 on lysine 27 (H3K27me3). The somatic mutation EZH2 Y641 in its highly conserved catalytic SET domain has been reported in diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma (FL) by methods of Sanger sequencing and whole exome sequencing. To develop a clinically applicable assay uses pyrosequencing method for accurate detection of EZH2Y641 mutations in formalinfixed-paraffin-embedded (FFPE) tissue. Methods: We validated the pyrosequencing method for the detection of EZH2 Y641 mutation in DLBCL cell lines. We also screened a cohort of 43 DLBCL and 12 FL cases by pyrosequencing, and correlated with the results of Sanger sequencing of