June 2020
Volume 61, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2020
ROLE OF CITICOLINE IN AN IN VITRO AMD MODEL
Author Affiliations & Notes
  • Sonali R Nashine
    University of California at Irvine, Irvine, California, United States
  • Cristina M Kenney
    University of California at Irvine, Irvine, California, United States
  • Footnotes
    Commercial Relationships   Sonali Nashine, None; Cristina Kenney, None
  • Footnotes
    Support  SN is a recipient of the 2017 Genentech/ ARVO AMD Translational Research Fellowship, RPB pilot research grant, and the Arnold and Mabel Beckman Postdoctoral fellowship.This work was supported by the Discovery Eye Foundation, Polly and Michael Smith, Edith and Roy Carver, Iris and B. Gerald Cantor Foundation, and NEI R01 EY0127363 (MCK). Supported in part by an Unrestricted Departmental Grant from Research to Prevent Blindness. We acknowledge the support of the Institute for Clinical and Translational Science (ICTS) at University of California Irvine.
Investigative Ophthalmology & Visual Science June 2020, Vol.61, 4436. doi:
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    • Get Citation

      Sonali R Nashine, Cristina M Kenney; ROLE OF CITICOLINE IN AN IN VITRO AMD MODEL. Invest. Ophthalmol. Vis. Sci. 2020;61(7):4436.

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

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Abstract

Purpose : In quest of identifying novel therapeutic candidates for Age-related Macular Degeneration (AMD), this study tested the hypothesis that Citicoline, a naturally occurring nootropic, will protect against apoptotic cell death in transmitochondrial AMD RPE cybrid cells (an in vitro AMD model). Citicoline maintains neuronal membrane integrity and its endogenous production from choline is an intermediate step in the synthesis of cell membrane phospholipids.

Methods : Purified Citicoline was exogenously added to transmitochondrial AMD RPE cybrid cells which had identical nuclei derived from mitochondria-deficient ARPE-19 cells, but differed in mitochondrial (mt) DNA content derived from clinically characterized AMD patients. The ability of Citicoline to attenuate apoptosis was examined via: 1) Analysis of untreated and Citicoline-treated AMD cybrid cells stained with Caspase-3/7 (quantifies cells undergoing caspase-3/7-mediated apoptosis) and NucLight red (stains live-cell nuclei) reagents in an IncuCyte live-cell imaging system; 2) Flow Cytometry analysis of untreated and Citicoline-treated AMD cybrid cells stained with apoptotic and dead cell markers namely AnnexinV and Propidium Iodide (PI) respectively.

Results : Addition of Citicoline led to a 55.99 % decrease in Overlap object count (i.e., Caspase-3/7 Green+NucLight Red staining)/ NucLight Red object count in AMD cybrid cells: Untreated- 1±0.078 a.u. and Citicoline-treated- 0.440±0.125 a.u.(p=0.03,n=4) at 48 h. At 72 h, a 47.54 % drop in Overlap object count was observed in Citicoline-treated AMD cybrid cells (0.52±0.11 a.u.) compared to their untreated counterparts (1±0.082 a.u.) (p=0.03, n=4). Furthermore, Flow cytometry analyses revealed a 21.67 % decrease in AnnexinV/PI double positives’ fluorescence intensity in Citicoline-treated AMD cybrid cells (0.783±0.06 a.u.) compared to their untreated counterparts (1±0.059 a.u.)(p=0.04, n=6).

Conclusions : In conclusion, this data support our hypothesis as Citicoline conferred significant protection against apoptotic cell death that was in-part mediated by damaged mtDNA from AMD patients in transmitochondrial AMD cybrid cells. Although further studies with Citicoline/ AMD cybrid cells are underway, these results present novel findings that identify Citicoline to be a potential protector which attenuates apoptotic cell death in AMD. Citicoline usage offers the advantage of easy access as it is available as an over-the-counter dietary supplement in the U.S.

This is a 2020 ARVO Annual Meeting abstract.

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