July 2019
Volume 60, Issue 9
Free
ARVO Annual Meeting Abstract  |   July 2019
Necroptosis in a murine repeated primary blast injury model
Author Affiliations & Notes
  • Chloe Naomi Thomas
    Neuroscience and Ophthalmology, Institute of Inflammation and Ageing, University of Birmingham, Edgbaston, Birmingham, England, United Kingdom
    Institute of Clinical Research, University of Birmingham, Birmingham, England, United Kingdom
  • Tonia S Rex
    Vanderbilt University Medical Center, Vanderbilt Eye Institute, Nashville, Tennessee, United States
  • Alexandra Bernardo-Colon
    Vanderbilt University Medical Center, Vanderbilt Eye Institute, Nashville, Tennessee, United States
  • Ella Courtie
    Neuroscience and Ophthalmology, Institute of Inflammation and Ageing, University of Birmingham, Edgbaston, Birmingham, England, United Kingdom
  • Gareth Essex
    Neuroscience and Ophthalmology, Institute of Inflammation and Ageing, University of Birmingham, Edgbaston, Birmingham, England, United Kingdom
  • Zubair Ahmed
    Neuroscience and Ophthalmology, Institute of Inflammation and Ageing, University of Birmingham, Edgbaston, Birmingham, England, United Kingdom
  • Richard J Blanch
    Academic Department of Military Surgery and Trauma, Royal Centre for Defence Medicine, Birmingham, England, United Kingdom
    Neuroscience and Ophthalmology, Institute of Inflammation and Ageing, University of Birmingham, Edgbaston, Birmingham, England, United Kingdom
  • Footnotes
    Commercial Relationships   Chloe Thomas, None; Tonia Rex, None; Alexandra Bernardo-Colon, None; Ella Courtie, None; Gareth Essex, None; Zubair Ahmed, None; Richard Blanch, None
  • Footnotes
    Support  Fight for Sight PhD Studentship (Grant code: 1560/1561). Tonia Rex: DoD W81XWH-15-1-0096 (TR) and W81XWH-17-2-0055 (TR), NEI R01 EY022349 (TR), NIA R01 NS094595 (TR), NEI P30EY008126 (VVRC), Research Prevent Blindness Unrestricted Funds (VEI), Potocsnak Family – CSC Research Fund (TR), Ayers Research Fund in Regenerative Visual Neuroscience (TR), Ret. Maj. General Stephen L. Jones, MD Fund (TR), Mark Pigott Fund (TR), and Vanderbilt University Medical Center Cell Imaging Shared Resource core facility (Clinical and Translational Science Award Grant UL1 RR024975 from National Center for Research Resources).
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 4404. doi:
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    • Get Citation

      Chloe Naomi Thomas, Tonia S Rex, Alexandra Bernardo-Colon, Ella Courtie, Gareth Essex, Zubair Ahmed, Richard J Blanch; Necroptosis in a murine repeated primary blast injury model. Invest. Ophthalmol. Vis. Sci. 2019;60(9):4404.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Repeated primary blast injury (rPBI) causes visual dysfunction and degeneration. Necroptosis (regulated necrosis) promotes neuronal death in trauma, neurodegeneration and in ocular disease and involves interactions between receptor interacting protein kinase (RIPK)-1,-3 and mixed lineage kinase domain-like (MLKL). RIPK1 and RIPK3 are elevated after a single 26 psi blast and necroptosis promotes retinal ganglion cell (RGC) death in blunt ocular injury. We aimed to determine if necroptosis promotes RGC death and ON degeneration in a murine rPBI model and if inhibition using Necrostatin-1 stable (Nec-1s) was protective.

Methods : We exposed the left eye of anesthetised C57Bl/6 mice to repeated blast of overpressure air (2x15 psi blasts in quick succession, 24 hours apart over 3 days). Sham mice were exposed to anaesthetics but not the blast wave. Nec-1s (2µl of 3.6mM Nec-1s in DMSO and 0.09% methyl-β-cyclodextrin) or vehicle control were intravitreally injected 48 hours before blast and repeated every 7 days and mice culled at 28 days post injury (dpi) (n=10 per group). Eyes were cryosectioned and RBPMS+ immunostained RGC quantified. Far proximal ON was resin embedded, sectioned, and stained with p-phenylenediamine and toluidine blue. Total healthy and degenerating axons were counted. The remaining ON was cryosectioned longitudinally and immunostained. Retinal optical coherence tomography (OCT) imaging was performed throughout the experiment and % of eyes with vitreal inflammation quantified.

Results : There were no differences in RBPMS+ RGC between sham and rPBI, but a reduction with combined rPBI and intravitreal injections. Nec-1s provided neuroprotection compared to vehicle (13.08 ± 0.81 compared to 10.29 ± 0.37 RBPMS+ cells/mm of retina). OCT images showed more vitreal inflammation in eyes receiving blast and injection combination (0% in sham and rPBI compared to 83% in vehicle and 88% in Nec-1s). The direct blast left eye had more inflammation compared to contralateral eye (33% in vehicle and 11% in Nec-1s). There were increased degenerating axons after rPBI which were not affected by Nec-1s. ED1+ macrophages in the ON increased after rPBI and remained elevated with Nec-1s.

Conclusions : Necroptosis might have a role in RGC death after rPBI. Intravitreal injections in combination with direct blast exposure increased vitreal inflammation and RGC death, suggesting an alternative administration for therapy would be preferable.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

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