MiRNA Profiling In Tumor Tissue, Body Fluids And Exosomes – A Combinational Techniques Approach Of NGS And QPCR.

Robert P. Loewe
GeneWake GmbH, Germany

miRNA has gained a pivotal role in molecular diagnostics and disease analysis. Due its stable nature and an abundant presence either in the tissue of origin, in microparticles or free circulating, it is a viable and interesting analyte. To understand the dynamics and spectrum of this biological dilution – from the production site down to circulation – we analysed glioblastoma tissue in conjunction with cerebrospinal fluid (CSF) and serum from the same patients. As the generation of microvesicles, general gene expression and epigenetic changes (methylation and hydroxyl-methylation of DNA) might mechanistically contribute to the phenomena, these characteristics were also measured. miRNA was primarily analysed via miRNA-Seq on a HiSeq instrument to allow a certain depth of the data. A data convergence of the different biological levels and source materials in addition to further information via qPCR was gathered. The profiling procedure and results underlining the inherent dynamics will be presented.

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