Lysine-specific histone demethylase 1A (KDM1A/LSD1) inhibition attenuates DNA double strand break repair and augments efficacy of temozolomide in glioblastoma II

Patient-derived primary GSCs were established from discarded specimens obtained from glioblastoma (GBM) patients undergoing surgery at UT Health San Antonio. We examined global transcriptional changes by RNA-sequencing using patient derived glioma stem cells (GSCs) treated with either vehicle, KDM1A inhibitor (NCD38) or temozolomide (TMZ) , or combination of NCD38+TMZ. Patient-derived GSCs (GSC082209) were treated with vehicle or NCD38 (3 µM) or TMZ (50 µM) or NCD38+TMZ for 24 hours. The RNA was isolated and utilized for RNA-seq analysis. Overall design: Patient derived GSC08 cells were treated with vehicle or NCD38 (3 µM) or TMZ (50 µM) or NCD38+TMZ for 24 hours. The RNA was isolated and utilized for RNA-seq analysis. Total RNA was isolated using RNeasy mini kit according to the manufacturer's instructions (Qiagen, Valencia, CA). Illumina TruSeq RNA Sample Preparation was performed following the manufacturer's protocol. Samples were run on an Illumina HiSeq 3000 in duplicate. The combined raw reads were aligned to UCSC hg19, and genes were annotated by Tophat. Genes were annotated and quantified by the HTSeq-DESeq pipeline.