title: A Customized Quantitative PCR MicroRNA Panel Provides a Technically Robust Context for Studying Neurodegenerative Disease Biomarkers and Indicates a High Correlation Between Cerebrospinal Fluid and Choroid Plexus MicroRNA Expression.
abstract: MicroRNA (miRNA) expression varies in association with different tissue types and in diseases. Having been found in body fluids including blood and cerebrospinal fluid (CSF), miRNAs constitute potential biomarkers. CSF miRNAs have been proposed as biomarkers for neurodegenerative diseases; however, there is a lack of consensus about the best candidate miRNA biomarkers and there has been variability in results from different research centers, perhaps due to technical factors. Here, we sought to optimize technical parameters for CSF miRNA studies. We examined different RNA isolation methods and performed miRNA expression profiling with TaqMan® miRNA Arrays. More specifically, we developed a customized CSF-miRNA low-density array (TLDA) panel that contains 47 targets: miRNAs shown previously to be relevant to neurodegenerative disease, miRNAs that are abundant in CSF, data normalizers, and controls for potential blood and tissue contamination. The advantages of using this CSF-miRNA TLDA panel include specificity, sensitivity, fast processing and data analysis, and cost effectiveness. We optimized technical parameters for this assay. Further, the TLDA panel can be tailored to other specific purposes. We tested whether the profile of miRNAs in the CSF resembled miRNAs isolated from brain tissue (hippocampus or cerebellum), blood, or the choroid plexus. We found that the CSF miRNA expression profile most closely resembles that of choroid plexus tissue, underscoring the potential importance of choroid plexus-derived signaling through CSF miRNAs. In summary, the TLDA miRNA array panel will enable evaluation and discovery of CSF miRNA biomarkers and can potentially be utilized in clinical diagnosis and disease stage monitoring.
authors.affiliation: Sanders Brown Center on Aging, University of Kentucky, Lexington, KY, 40536, USA. wwangc@email.uky.edu.
authors.forename: Wang-Xia
authors.identifier: http://orcid.org/0000-0002-8104-3779
authors.initials: WX
authors.lastname: Wang
authors.affiliation: Sanders Brown Center on Aging, University of Kentucky, Lexington, KY, 40536, USA.|Department of Biostatistics, University of Kentucky, Lexington, KY, 40536, USA.
authors.forename: David W
authors.initials: DW
authors.lastname: Fardo
authors.affiliation: Sanders Brown Center on Aging, University of Kentucky, Lexington, KY, 40536, USA.|Department of Neurology, University of Kentucky, Lexington, KY, 40536, USA.
authors.forename: Gregory A
authors.initials: GA
authors.lastname: Jicha
authors.affiliation: Sanders Brown Center on Aging, University of Kentucky, Lexington, KY, 40536, USA. pnels2@email.uky.edu.|Department of Pathology, Division of Neuropathology, University of Kentucky, Lexington, KY, 40536, USA. pnels2@email.uky.edu.
authors.forename: Peter T
authors.initials: PT
authors.lastname: Nelson
mesh_terms: D015415:Biomarkers / Q000134:cerebrospinal fluid / Q000378:metabolism; D001921:Brain / Q000378:metabolism; D002831:Choroid Plexus / Q000378:metabolism*; D015870:Gene Expression; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D035683:MicroRNAs / Q000096:biosynthesis / Q000235:genetics*; D019636:Neurodegenerative Diseases / Q000134:cerebrospinal fluid* / Q000235:genetics* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards
chemical_list: D015415:Biomarkers; D035683:MicroRNAs
keywords: Biofluid; Biomarker; Cerebrospinal fluid; Choroid plexus; MicroRNA; Neurodegenerative disease