Genetic Profiling of Meningioma

Variants identified by WES, as well as selected regions of previously published genes implicated in meningioma (NF2, AKT1, TRAF7, SMO, and KLF4), were analyzed by PCR and Sanger sequencing. Mutant sites from NF2 (22), TRAF7 (5), AKT1 (1), SMO (2), KLF4 (1), and TERT (2) were sequenced. DNA from the four meningioma cell lines and one normal human cell line (293T) were amplified using PCR primers designed to target identified mutations (S1 Table). PCR products were separated by 1% agarose gel, visualized under UV light, extracted using the Qiagen Gel Extraction Kit (Valencia, CA), and subject to Sanger sequencing (Macrogen, Boston, MA). Mutant variants identified by Sanger sequencing were verified by alignment of sequenced results with sequences of normal human cell line 293T and human genomic DNA reference (hg19). Furthermore, the TERT promoter region was focally sequenced across cell lines to assess for presence of TERT mutations, given the important role it may play in meningioma progression [7 (link), 21 (link)].