June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Histologic cell shape descriptors for the retinal pigment epithelium in age-related macular degeneration
Author Affiliations & Notes
  • Leon Alexander von der Emde
    Ophthalmology, University eye hospital in Bonn, Bonn, NRW, Germany
  • Marc Vaisband
    University of Bonn, Life & Medical Sciences Institute, Bonn, Germany, Germany
    Department of Internal Medicine III with Haematology, Medical Oncology, Haemostaseology, Infectiology and Rheumatology, Oncologic Center; Salzburg Cancer Research Institute - Laboratory for Immunological and Molecular Cancer Research (SCRI-LIMCR); Paracelsus Medical University, Salzburg, Austria, Cancer Cluster Salzburg, Austria, Austria
  • Jan Hasenauer
    University of Bonn, Life & Medical Sciences Institute, Bonn, Germany, Germany
    Helmholtz Center Munich- German Research Center for Environmental Health, Institute of Computational Biology, Neuherberg, Germany, Germany
  • Leonie Bourauel
    Ophthalmology, University eye hospital in Bonn, Bonn, NRW, Germany
  • Marlene Sassmannshausen
    Ophthalmology, University eye hospital in Bonn, Bonn, NRW, Germany
  • Katharina Bermond
    Department of Ophthalmology, Ludwigshafen Hospital, Ludwigshafen, Germany, Germany
  • Rainer Heintzmann
    eibniz Institute of Photonic Technology, Jena, Germany, Germany
    Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich-Schiller University Jena, Jena, Germany, Germany
  • Frank G Holz
    Ophthalmology, University eye hospital in Bonn, Bonn, NRW, Germany
  • Christine A Curcio
    Department of Ophthalmology and Visual Sciences, University of Alabama at Birmingham, Alabama, AL, United States, Alabama, United States
  • Kenneth R Sloan
    Department of Ophthalmology and Visual Sciences, University of Alabama at Birmingham, Alabama, AL, United States, Alabama, United States
  • Thomas Ach
    Ophthalmology, University eye hospital in Bonn, Bonn, NRW, Germany
  • Footnotes
    Commercial Relationships   Leon von der Emde None; Marc Vaisband None; Jan Hasenauer None; Leonie Bourauel None; Marlene Sassmannshausen Heidelberg Engineering, Code F (Financial Support), Carl Zeiss Meditec, Code F (Financial Support), Centervue, Code F (Financial Support); Katharina Bermond None; Rainer Heintzmann Zeiss, Code C (Consultant/Contractor), Zeiss, Code F (Financial Support); Frank Holz Acucela, Code C (Consultant/Contractor), Appellis, Code C (Consultant/Contractor), Bayer, Code C (Consultant/Contractor), Boehringer-Ingelheim, Code C (Consultant/Contractor), Bioeq/Formycon , Code C (Consultant/Contractor), Roche, Code C (Consultant/Contractor), Acucela, Code F (Financial Support), Allergan, Code F (Financial Support); Christine Curcio Genentech, Code F (Financial Support), Heidelberg Engineering, Code F (Financial Support), Regeneron, Code F (Financial Support), MacRegen Inc., Code I (Personal Financial Interest); Kenneth Sloan MacRegen Inc., Code I (Personal Financial Interest); Thomas Ach Roche, Code C (Consultant/Contractor), Novartis, Code C (Consultant/Contractor), MacRegen Inc., Code I (Personal Financial Interest), Novartis, Code R (Recipient)
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 4508 – F0295. doi:
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      Leon Alexander von der Emde, Marc Vaisband, Jan Hasenauer, Leonie Bourauel, Marlene Sassmannshausen, Katharina Bermond, Rainer Heintzmann, Frank G Holz, Christine A Curcio, Kenneth R Sloan, Thomas Ach; Histologic cell shape descriptors for the retinal pigment epithelium in age-related macular degeneration. Invest. Ophthalmol. Vis. Sci. 2022;63(7):4508 – F0295.

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

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Abstract

Purpose : Phenotypic alterations of retinal pigment epithelium (RPE) cells are central in age-related macular degeneration (AMD). Shape descriptors for individual RPE cells based on RPEs cytoskeleton may help to delineate healthy from AMD affected cells, even in early stages of disease. Here, we quantified differences in RPE morphology between donors with unaffected macula and with AMD.

Methods : Twenty-two human RPE flatmounts (7 AMD, mean age 85 ± 3 yrs (early: 3, geographic atrophy: 1; neovascular: 3;); 15 unaffected (≤51 yrs, n = 8; >80 yrs, n = 7)) were imaged at different locations (fovea, perifovea, near periphery) using a laser scanning confocal fluorescence microscope (exc. 488 nm; emission: 490-695 nm; z-stack: 390 nm steps). Hypoautofluorescent gaps between adjacent RPE cells were formed by the absence of lipofuscin/melanolipofuscin granules between the F-actin cytoskeletons. These gaps were manually marked with computer-assistance and considered borders between cells. Shape descriptors including form factor (measure for circularity), area, solidity, convexity, and roundness for each cell were calculated using customized software. Total autofluorescence (AF) per cell was defined as the integrated density of intensity from all pixels within the cell. Statistical analysis was performed using an ensemble classifier based on logistic regression.

Results : Comparing young vs healthy aged eyes, there was a trend to increased area and reduced form factor. In AMD eyes, RPE shape was significantly altered at all locations with area, solidity and form factor being the most discriminatory descriptors. In samples from healthy donors, with increasing distance to the fovea, area, solidity, and convexity increased, while form factor decreased. Finally, reduced RPE cell AF in AMD was significantly associated with decreased roundness and solidity of the RPE.

Conclusions : Morphological RPE cell alterations in presence of AMD can be accurately quantified using cell shape descriptors. Enlarged and deformed cells in AMD are assumed to be indicative of structurally and functionally impaired RPE cells. The results of this study may help guide the interpretation of RPE morphology in in vivo studies utilizing high-resolution single cell imaging, e.g., adaptive optics scanning laser ophthalmoscopes or transscleral optic phase imaging.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

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