June 2020
Volume 61, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2020
RPE cellular morphology and intracellular autofluorescence after suppression of vitreous complement levels
Author Affiliations & Notes
  • Anthony Jones
    Ophthalmology, CU Anschutz, Aurora, Colorado, United States
  • Joshua Logan Morgenstern
    Ophthalmology, CU Anschutz, Aurora, Colorado, United States
  • Jeffrey L Olson
    Ophthalmology, CU Anschutz, Aurora, Colorado, United States
  • Footnotes
    Commercial Relationships   Anthony Jones, None; Joshua Morgenstern, None; Jeffrey Olson, 2C Tech equity (F), Genentech clinical reset funding (F), Patent application via CU Board of Regents for this technology (P)
  • Footnotes
    Support  Gates Grubstake Fund
Investigative Ophthalmology & Visual Science June 2020, Vol.61, 318. doi:
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      Anthony Jones, Joshua Logan Morgenstern, Jeffrey L Olson; RPE cellular morphology and intracellular autofluorescence after suppression of vitreous complement levels. Invest. Ophthalmol. Vis. Sci. 2020;61(7):318.

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

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Abstract

Purpose : Age-related macular degeneration (AMD) is associated with changes in the retinal pigmented epithelial (RPE) cells that can be monitored clinically through autofluorescence (AF). Most of these pathologic changes are related to RPE lipofuscin which can be seen on a cellular level as aggregates and pre-aggregates. Furthermore, cellular morphology has been shown to be an indicator of disease progression. In this study we analyzed both cellular AF and cellular morphology in a mouse AMD model and assessed the effect a polyacylonitrile (PAN) implant has on disease progression.

Methods : Three CFH -/- mice, with known abnormal complement activation and subsequent photoreceptor dysfunction, received intravitreal PAN injections in the right eye (n=3) while the left eye (n=3) served as the control. 5 months post injection, eyes were enucleated and prepared for flat mounts with the tissue cytoskeleton labeled with 647 Alexa-conjugated phalloidin. Cell morphology and AF were assessed using a previously established classification system specific for RPE cells. Morphological derangement was determined on the prevalence of misshapen cell boundaries, while AF was based on signal intensity and amount of granule aggregates and pre-aggregates after exposure to 488nm. Statistical analysis was performed using a two tailed student's t-test.

Results : Cytoskeletal derangements differing from the healthy polygonal cellular morphology range from round, mixed and concave, and were classified as misshapen. Fewer aberrations were seen in the PAN treatment groups with a statistically significant difference between all study subjects (p<0.05, t-test). Compared to the control group, the PAN treated eyes displayed significantly less (p<0.05) AF intensity and granule aggregates/pre-aggregates.

Conclusions : PAN implanted eyes showed cellular anatomical preservation with less RPE dysmorphia. Furthermore, the treatment eyes had significantly less AF aggregates and pre-aggregates than the non-PAN implant eyes. These findings elucidate on the utility of PAN as a novel therapy for the attenuation of AMD pathology as seen form a cellular level. Our results offer insight into an additional avenue for management of dry AMD, a disease with currently no treatment options.

This is a 2020 ARVO Annual Meeting abstract.

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