10779/rcsi.10790735.v1 Rana Raoof Rana Raoof Sebastian Bauer Sebastian Bauer Hany El Naggar Hany El Naggar Gary P Brennan Gary P Brennan Elizabeth Brindley Elizabeth Brindley Thomas Hill Thomas Hill Hazel McArdle Hazel McArdle Elaine Spain Elaine Spain Robert J Forster Robert J Forster Jochen HM Prehn Jochen HM Prehn Hajo Hamer Hajo Hamer Norman Delanty Norman Delanty Felix Rosenow Felix Rosenow Catherine Mooney Catherine Mooney David C. Henshall David C. Henshall Dual-center, dual-platform microRNA profiling identifies potential plasma biomarkers of adult temporal lobe epilepsy. Royal College of Surgeons in Ireland 2019 Animals Biomarkers Case-Control Studies Circulating MicroRNA Computational Biology Disease Models Animal Epilepsy Temporal Lobe Gene Expression Profiling Gene Regulatory Networks High-Throughput Nucleotide Sequencing Humans Mice MicroRNAs Transcriptome Physiology Medical Physics 2019-11-22 16:56:28 Journal contribution https://repository.rcsi.com/articles/journal_contribution/Dual-center_dual-platform_microRNA_profiling_identifies_potential_plasma_biomarkers_of_adult_temporal_lobe_epilepsy_/10790735 <p>BACKGROUND: There are no blood-based molecular biomarkers of temporal lobe epilepsy (TLE) to support clinical diagnosis. MicroRNAs are short noncoding RNAs with strong biomarker potential due to their cell-specific expression, mechanistic links to brain excitability, and stable detection in biofluids. Altered levels of circulating microRNAs have been reported in human epilepsy, but most studies collected samples from one clinical site, used a single profiling platform or conducted minimal validation.</p> <p>METHOD: Using a case-control design, we collected plasma samples from video-electroencephalogram-monitored adult TLE patients at epilepsy specialist centers in two countries, performed genome-wide PCR-based and RNA sequencing during the discovery phase and validated findings in a large (>250) cohort of samples that included patients with psychogenic non-epileptic seizures (PNES).</p> <p>FINDINGS: After profiling and validation, we identified miR-27a-3p, miR-328-3p and miR-654-3p with biomarker potential. Plasma levels of these microRNAs were also changed in a mouse model of TLE but were not different to healthy controls in PNES patients. We determined copy number of the three microRNAs in plasma and demonstrate their rapid detection using an electrochemical RNA microfluidic disk as a prototype point-of-care device. Analysis of the microRNAs within the exosome-enriched fraction provided high diagnostic accuracy while Argonaute-bound miR-328-3p selectively increased in patient samples after seizures. In situ hybridization localized miR-27a-3p and miR-328-3p within neurons in human brain and bioinformatics predicted targets linked to growth factor signaling and apoptosis.</p> <p>INTERPRETATION: This study demonstrates the biomarker potential of circulating microRNAs for epilepsy diagnosis and mechanistic links to underlying pathomechanisms.</p>