Assessing Neuronal and Astrocyte Derived Exosomes From Individuals With Mild Traumatic Brain Injury for Markers of Neurodegeneration and Cytotoxic Activity

Charisse N Winston; Haylie K Romero; Maya Ellisman; Sophie Nauss; David A Julovich; Tori Conger; James R Hall; Wendy Campana; Sid E O'Bryant; Caroline M Nievergelt; Dewleen G Baker; Victoria B Risbrough; Robert A Rissman

doi:10.3389/fnins.2019.01005

Assessing Neuronal and Astrocyte Derived Exosomes From Individuals With Mild Traumatic Brain Injury for Markers of Neurodegeneration and Cytotoxic Activity

Front Neurosci. 2019 Oct 2:13:1005. doi: 10.3389/fnins.2019.01005. eCollection 2019.

Authors

Charisse N Winston¹, Haylie K Romero², Maya Ellisman¹, Sophie Nauss¹, David A Julovich³, Tori Conger³, James R Hall³, Wendy Campana^{2

4}, Sid E O'Bryant³, Caroline M Nievergelt^{5

6}, Dewleen G Baker^{5

6}, Victoria B Risbrough^{5

6}, Robert A Rissman^{1

4}

Affiliations

¹ Department of Neurosciences, University of California, San Diego, La Jolla, CA, United States.
² Department of Anesthesiology, University of California, San Diego, La Jolla, CA, United States.
³ Department of Pharmacology and Neuroscience, Institute for Translational Research, University of North Texas Health Science Center, Fort Worth, TX, United States.
⁴ VA San Diego Healthcare System, La Jolla, CA, United States.
⁵ Center of Excellence for Stress and Mental Health, La Jolla, CA, United States.
⁶ Department of Psychiatry, University of California, San Diego, La Jolla, CA, United States.

Abstract

Mild traumatic brain injury (mTBI) disproportionately affects military service members and is very difficult to diagnose. To-date, there is currently no blood-based, diagnostic biomarker for mTBI cases with persistent post concussive symptoms. To examine the potential of neuronally-derived (NDE) and astrocytic-derived (ADE) exosome cargo proteins as biomarkers of chronic mTBI in younger adults, we examined plasma exosomes from a prospective longitudinal study of combat-related risk and resilience, marine resiliency study II (MRSII). After return from a combat-deployment participants were interviewed to assess TBI exposure while on deployment. Plasma exosomes from military service members with mTBI (mean age, 21.7 years, n = 19, avg. days since injury 151), and age-matched, controls (deployed service members who did not endorse a deployment-related TBI or a pre-deployment history of TBI; mean age, 21.95 years, n = 20) were precipitated and enriched against a neuronal adhesion protein, L1-CAM, and an astrocyte marker, glutamine aspartate transporter (GLAST) using magnetic beads to immunocapture the proteins and subsequently selected by fluorescent activated cell sorting (FACS). Extracted protein cargo from NDE and ADE preparations were quantified for protein levels implicated in TBI neuropathology by standard ELISAs and on the ultra-sensitive single molecule assay (Simoa) platform. Plasma NDE and ADE levels of Aβ42 were significantly higher while plasma NDE and ADE levels of the postsynaptic protein, neurogranin (NRGN) were significantly lower in participants endorsing mTBI exposure compared to controls with no TBI history. Plasma NDE and ADE levels of Aβ40, total tau, and neurofilament light (NFL), P-T181-tau, P-S396-tau were either undetectable or not significantly different between the two groups. In an effort to understand the pathogenetic potential of NDE and ADE cargo proteins, neuron-like cultures were treated with NDE and ADE preparations from TBI and non-TBI groups. Lastly, we determined that plasma NDE but not ADE cargo proteins from mTBI samples were found to be toxic to neuron-like recipient cells in vitro. These data support the presence of markers of neurodegeneration in NDEs of mTBI and suggest that these NDEs can be used as tools to identify pathogenic mechanisms of TBI.

Keywords: amyloid; astrocytes; neuronal exosomes; tau; traumatic brain injury.

Abstract

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