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Serum is preferable to plasma for circulating exosomal microRNA biomarker discovery studies

ABSTRACT: Exosomes were purified from 250 ul matched plasma and serum samples using ExoQuickTm. The presence of particles consistent in size with exosomes (60-150nm) was confirmed using a Nanosight LM10. miRNA was extracted from exosomes using an miRNeasy Serum/Plasma kit (Qiagen, #217184). miRNA was reversed transcribed using a TaqMan® microRNA Reverse Transcription Kit (Life technologies, #4366596). miRNA profiling was performed with a high throughput TaqMan® OpenArray® Human microRNA panel (Life technologies, #4461104). The panel consisted of probes for 754 human miRNAs that are based on miRNA sequences derived from Sanger miRBase v14. MegaplexTM Primer Human Pool A v2.1 and Human Pool B v2.0 or v3.0

ORGANISM(S): Homo sapiens

PROVIDER: GSE142855 | GEO | 2022/09/01

REPOSITORIES: GEO

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Circulating serum miRNAs as potential diagnostic biomarkers for oesophageal adenocarcinoma

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Nanostring human miRNA panel: exosome fraction from peripheral blood of trauma patients versus healthy

Project description:827 human miRNA standard human panel Plasma derived extracellular vesicles (EV)/exosomes can serve as markers of cell damage/disease but can also have therapeutic utility depending on the nature of their cargo, such as miRNA. Currently there are challenges and lack of innovations regarding early diagnosis and therapeutic options within different aspects of management of trauma patients. We characterized the miRNA cargo of exosomes purified from trauma patients, and compared it to those from healthy participants. Methods Plasma was collected from a prospective, cohort study of trauma patients (n = 15; 7 males, 8 females) with hip and/or femur fractures and an injury severity score (ISS) ≥ 15; elective total hip arthroplasty (THA) patients (n = 8; 4 males, 4 females) served as operative controls. Exosomes were isolated from plasma with the Invitrogen Total Exosome Isolation Kit and RNA was isolated using a miRNeasy Mini Kit. Direct quantification of miRNA copies was performed by NanoString Technologies on a human miRNA gene panel and analyzed with nSolver Results Trauma plasma-derived exosomes had 60 miRNA that were significantly downregulated and 3 miR upregulated when compared to THA (p<0.05). Thirteen miR have a direct role in hematopoiesis regulation, including miR-223 and miR-451a. Further, male trauma plasma-derived exosomes demonstrated downregulation of 150 miR compared to male THA (p<0.05). While female trauma plasma-derived exosomes demonstrated downregulation of only four miR and upregulation of 2 miR compared to female THA (p<0.05). Conclusions Sex-specific differences in the miR from plasma-derived exosomes occurred following severe trauma. These miR changes in plasma-derived exosomes likely contribute to postinjury systemic inflammation, immune system dysregulation, and bone marrow dysfunction and may merit consideration as future potential therapeutic targets.

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