Targeted RNASeq using a custom panel of metabolic genes

Purpose: The purpose of this experiment was to determine the time point following activation at which greatest transcriptional differences in metabolic genes were observed between primary human CD4+ T cells treated with metformin + 2-deoxyglucose combination regimen and cells treated with vehicle. Methods: Targeted RNA-Seq data was generated from primary human CD4+ T cell mRNA using a Qiagen custom designed panel of metabolic genes and the Illumina TruSeq stranded mRNA prep. Results: We obtained differential gene expression between metformin + 2-deoxyglucose and vehicle-treated cells using the DESeq2 app in Illumina Basespace for a set of metabolic genes during different time points following activation, and found the greatest changes at 24hr. Overall design: Targeted RNASeq transcriptomic profiling of primary human CD4+ T cells activated for 0, 3, 6, 9, and 24 hr with soluble anti-CD3 and anti-CD28 tetrameric complexes (Immunocult) in the presence of metformin and 2-deoxyglucose, of 2 individual donors.