June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Gene-environment interaction between mitophagy regulator, Pink1, and smoking generates a range of AMD like phenotype in the mouse RPE
Author Affiliations & Notes
  • Sayantan Datta
    Ophthalmology, Emory University School of Medicine, Atlanta, Georgia, United States
  • GANESH SATYANARAYANA
    Ophthalmology, Emory University School of Medicine, Atlanta, Georgia, United States
  • Marisol Cano
    Ophthalmology, Johns Hopkins Medicine, Baltimore, Maryland, United States
  • James T Handa
    Ophthalmology, Johns Hopkins Medicine, Baltimore, Maryland, United States
  • Footnotes
    Commercial Relationships   Sayantan Datta None; GANESH SATYANARAYANA None; Marisol Cano None; James Handa Character Biosciences, Seeing Medicines Cirrus Therapeutics, , Code C (Consultant/Contractor)
  • Footnotes
    Support  NEI/NIH R00EY029010
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 2303. doi:
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      Sayantan Datta, GANESH SATYANARAYANA, Marisol Cano, James T Handa; Gene-environment interaction between mitophagy regulator, Pink1, and smoking generates a range of AMD like phenotype in the mouse RPE. Invest. Ophthalmol. Vis. Sci. 2023;64(8):2303.

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

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Abstract

Purpose : Dry AMD is a complex disease exhibiting a great degree of heterogeneity, which is both poorly modelled and understood. In AMD, mitochondrial dysfunction is an important genetic factor and smoking is the strongest environmental risk factor, however the joint effect of these two risk factors have not been investigated in the context of AMD. Dysfunction and dysmorphology of the retinal pigment epithelial (RPE) cells is an early feature in AMD and here we investigate the effect of Pink1 loss, the master regulator of mitophagy and smoking on RPE structure and function in Pink1-/- mice. In AMD, RPE cells show both EMT (epithelial to mesenchymal transition) and cell death, decrease in expression of RPE specific markers and overall structural and functional decline.

Methods : Four-month-old C57 and Pink1-/- mice were smoked on Kentucky Reference Cigarettes or on air in a smoking chamber for 2hrs/day, 5days/week for 6months and evaluated for: i. RPE morphology and shape using RPE flatmounts, ii. Sub-cellular deposits and RPE inclusions using TEM, iii. Gene expression of RPE specific genes – Lrat, Best1, Rpe65 using qPCR and western blots, iv. Gene expression of EMT (e-Cadherin, Vimentin, Snail1) and cell death (Bcl-2, Caspase-3,9) related genes using qPCR and western blots, v. Functional evaluation using scotopic electroretinogram (ERG).

Results : Pink1-/- mice even without smoking showed dysmorphic RPE with typical EMT morphology and 2-fold increase in EMT markers (P<0.005 for all markers), basal laminar deposits and significantly decreased ERG b and c waves compared to C57 (both air and smoked) mice. However, Pink1-/- mice which were smoked showed evidence of increased cell death both morphologically (non-nucleated, vacuole filled RPE cells in TEM) and biochemically (increased caspase expression) and showed more pronounced basal deposits. ERG a, b and c waves were significantly decreased compared to both C57 smoked and Pink1 air-raised mice. C57 smoked mice compared to C57 air mice showed slight increase in EMT markers and some evidence of basal deposits but not as strong as those seen in Pink1-/- mice.

Conclusions : Thus, Pink1-/- mimics dysfunctional RPE which are still surviving undergoing EMT whereas an added environmental insult drives the cells towards cell death. Our model helps capture the typical heterogeneity seen in the RPE of AMD patients.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

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