Monitoring of Post-Brain Injuries By Measuring Plasma Levels of Neuron-Derived Extracellular Vesicles

Naoshi Hotta; Takahiro Tadokoro; John Henry; Daisuke Koga; Keisuke Kawata; Hiroyuki Ishida; Yuko Oguma; Akihiro Hirata; Masato Mitsuhashi; Kenji Yoshitani

doi:10.1177/11772719221128145

Monitoring of Post-Brain Injuries By Measuring Plasma Levels of Neuron-Derived Extracellular Vesicles

Biomark Insights. 2022 Oct 26:17:11772719221128145. doi: 10.1177/11772719221128145. eCollection 2022.

Authors

Affiliations

¹ Department of Anesthesiology, National Cerebral and Cardiovascular Center, Osaka, Japan.
² NanoSomiX, Inc., Irvine, CA, USA.
³ Department of Anatomy, Asahikawa Medical University, Asahikawa, Japan.
⁴ School of Public Health, Indiana University, Bloomington, IN, USA.
⁵ Sports Medicine Research Center, Keio University, Kanagawa, Japan.

Abstract

Background: Extracellular vesicles (EV) released from neurons into the blood can reflect the state of nervous tissue. Measurement of neuron derived EV (NDE) may serve as an indicator of brain injury.

Methods: A sandwich immunoassay was established to measure plasma NDE using anti-neuron CD171 and anti-EV CD9 ([CD171 ⁺ CD9⁺]). Plasma samples were obtained from commercial sources, cross-country (n = 9), football (n = 22), soccer (n = 19), and rugby (n = 18) athletes over time. Plasma was also collected from patients undergoing total aortic arch replacement (TAR) with selective cerebral perfusion during cardiopulmonary bypass before and after surgery (n = 36).

Results: The specificity, linearity, and reproducibility of NDE assay (measurement of [CD171 ⁺ CD9⁺]) were confirmed. By scanning electron microscopy and nanoparticle tracking, spherical vesicles ranging in size from 150 to 300 nm were confirmed. Plasma levels of NDE were widely spread over 2 to 3 logs in different individuals with a significant age-dependent decrease. However, NDE were very stable in each individual within a ± 50% change over time (cross-country, football, soccer), whereas rugby players were more variable over 4 years. In patients undergoing TAR, NDE increased rapidly in days post-surgery and were significantly (P = .0004) higher in those developing postoperative delirium (POD) (n = 13) than non-delirium patients (n = 23).

Conclusions: The blood test to determine plasma levels of NDE was established by a sandwich immunoassay using 2 antibodies against neuron (CD171) and exosomes (CD9). NDE levels varied widely in different individuals and decreased with age, indicating that NDE levels should be considered as a normalizer of NDE biomarker studies. However, NDE levels were stable over time in each individual, and increased rapidly after TAR with greater increases associated with patients developing POD. This assay may serve as a surrogate for evaluating and monitoring brain injuries.

Keywords: CD171; CD9; ELISA; Extracellular vesicles; brain injuries; sports concussion; total aortic arch replacement.