title: miRNA contents of cerebrospinal fluid extracellular vesicles in glioblastoma patients.
abstract: Analysis of extracellular vesicles (EVs) derived from plasma or cerebrospinal fluid (CSF) has emerged as a promising biomarker platform for therapeutic monitoring in glioblastoma patients. However, the contents of the various subpopulations of EVs in these clinical specimens remain poorly defined. Here we characterize the relative abundance of miRNA species in EVs derived from the serum and cerebrospinal fluid of glioblastoma patients. EVs were isolated from glioblastoma cell lines as well as the plasma and CSF of glioblastoma patients. The microvesicle subpopulation was isolated by pelleting at 10,000×g for 30 min after cellular debris was cleared by a 2000×g (20 min) spin. The exosome subpopulation was isolated by pelleting the microvesicle supernatant at 120,000×g (120 min). qRT-PCR was performed to examine the distribution of miR-21, miR-103, miR-24, and miR-125. Global miRNA profiling was performed in select glioblastoma CSF samples. In plasma and cell line derived EVs, the relative abundance of miRNAs in exosome and microvesicles were highly variable. In some specimens, the majority of the miRNA species were found in exosomes while in other, they were found in microvesicles. In contrast, CSF exosomes were enriched for miRNAs relative to CSF microvesicles. In CSF, there is an average of one molecule of miRNA per 150-25,000 EVs. Most EVs derived from clinical biofluids are devoid of miRNA content. The relative distribution of miRNA species in plasma exosomes or microvesicles is unpredictable. In contrast, CSF exosomes are the major EV compartment that harbor miRNAs.
authors.affiliation: Center for Theoretical and Applied Neuro-Oncology, University of California, San Diego, CA, USA.
authors.forename: Johnny C
authors.initials: JC
authors.lastname: Akers
authors.forename: Valya
authors.initials: V
authors.lastname: Ramakrishnan
authors.forename: Ryan
authors.initials: R
authors.lastname: Kim
authors.forename: Shirley
authors.initials: S
authors.lastname: Phillips
authors.forename: Vivek
authors.initials: V
authors.lastname: Kaimal
authors.forename: Ying
authors.initials: Y
authors.lastname: Mao
authors.forename: Wei
authors.initials: W
authors.lastname: Hua
authors.forename: Isaac
authors.initials: I
authors.lastname: Yang
authors.forename: Chia-Chun
authors.initials: CC
authors.lastname: Fu
authors.forename: John
authors.initials: J
authors.lastname: Nolan
authors.forename: Ichiro
authors.initials: I
authors.lastname: Nakano
authors.forename: Yuanfan
authors.initials: Y
authors.lastname: Yang
authors.forename: Martin
authors.initials: M
authors.lastname: Beaulieu
authors.forename: Bob S
authors.initials: BS
authors.lastname: Carter
authors.forename: Clark C
authors.initials: CC
authors.lastname: Chen
mesh_terms: D015415:Biomarkers / Q000134:cerebrospinal fluid*; D000067128:Extracellular Vesicles / Q000235:genetics*; D020869:Gene Expression Profiling; D005909:Glioblastoma / Q000134:cerebrospinal fluid / Q000235:genetics*; D006801:Humans; D035683:MicroRNAs / Q000134:cerebrospinal fluid / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
chemical_list: D015415:Biomarkers; D035683:MicroRNAs; D012333:RNA, Messenger