Retinal Debris Triggers Cytotoxic Damage in Cocultivated Primary Porcine RPE Cells

Overview

Journal Front Neurosci

Date 2024 Aug 8

PMID 39114482

Authors

Natalie Wagner

Teresa Tsai

Sabrina Reinehr

Janine Theile

H Burkhard Dick

Stephanie C Joachim

Affiliations

Soon will be listed here.

Abstract

Introduction: One of the most common causes of vision loss in the elderly population worldwide is age-related macular degeneration (AMD). Subsequently, the number of people affected by AMD is estimated to reach approximately 288 million by the year 2040. The aim of this study was to develop an model that simulates various aspects of the complex AMD pathogenesis.

Methods: For this purpose, primary porcine retinal pigment epithelial cells (ppRPE) were isolated and cultured. One group was exposed to medium containing sodium iodate (NaIO) to induce degeneration. The others were exposed to different supplemented media, such as bovine serum albumin (BSA), homogenized porcine retinas (HPR), or rod outer segments (ROOS) for eight days to promote retinal deposits. Then, these ppRPE cells were cocultured with porcine neuroretina explants for another eight days. To assess the viability of ppRPE cells, live/dead assay was performed at the end of the study. The positive RPE65 and ZO1 area was evaluated by immunocytochemistry and the expression of , , and was analyzed by RT-qPCR. Additionally, drusen (), inflammation (, , , , ), oxidative stress (, , ), and hypoxia () markers were investigated. The concentration of the inflammatory cytokines IL-6 and IL-8 was determined in medium supernatants from day 16 and 24 via ELISA.

Results: Live/dead assay suggests that especially exposure to NaIO and HPR induced damage to ppRPE cells, leading in a significant ppRPE cell loss. All supplemented media resulted in decreased RPE-characteristic markers (RPE65; ZO-1) and gene expression like and in the cultured ppRPE cells. Besides, some inflammatory, oxidative as well as hypoxic stress markers were altered in ppRPE cells cultivated with NaIO. The application of HPR induced an enhanced expression. Pre-exposure of the ppRPE cells led to a diminished number of cones in all supplemented media groups compared to controls.

Discussion: Overall, this novel coculture model represents an interesting initial approach to incorporating deposits into coculture to mimic AMD pathogenesis. Nevertheless, the effects of the media used need to be investigated in further studies.

References

Tarallo V, Hirano Y, Gelfand B, Dridi S, Kerur N, Kim Y . DICER1 loss and Alu RNA induce age-related macular degeneration via the NLRP3 inflammasome and MyD88. Cell. 2012; 149(4):847-59. PMC: 3351582. DOI: 10.1016/j.cell.2012.03.036. View

Farjood F, Vargis E . Physical disruption of cell-cell contact induces VEGF expression in RPE cells. Mol Vis. 2017; 23:431-446. PMC: 5524271. View

Zarbin M . Current concepts in the pathogenesis of age-related macular degeneration. Arch Ophthalmol. 2004; 122(4):598-614. DOI: 10.1001/archopht.122.4.598. View

Page-McCaw A, Ewald A, Werb Z . Matrix metalloproteinases and the regulation of tissue remodelling. Nat Rev Mol Cell Biol. 2007; 8(3):221-33. PMC: 2760082. DOI: 10.1038/nrm2125. View

Wong W, Su X, Li X, Cheung C, Klein R, Cheng C . Global prevalence of age-related macular degeneration and disease burden projection for 2020 and 2040: a systematic review and meta-analysis. Lancet Glob Health. 2014; 2(2):e106-16. DOI: 10.1016/S2214-109X(13)70145-1. View

Wagner N, Safaei A, Vogt P, Gammel M, Dick H, Schnichels S . Coculture of ARPE-19 Cells and Porcine Neural Retina as an Retinal Model. Altern Lab Anim. 2022; 50(1):27-44. DOI: 10.1177/02611929221082662. View

Xie J, Zhu R, Peng Y, Gao W, Du J, Zhao L . Tumor necrosis factor-alpha regulates photoreceptor cell autophagy after retinal detachment. Sci Rep. 2017; 7(1):17108. PMC: 5719449. DOI: 10.1038/s41598-017-17400-3. View

Wagner N, Reinehr S, Gammel M, Greulich A, Hurst J, Dick H . Novel Porcine Retina Cultivation Techniques Provide Improved Photoreceptor Preservation. Front Neurosci. 2020; 14:556700. PMC: 7573241. DOI: 10.3389/fnins.2020.556700. View

Klein R, Myers C, Cruickshanks K, Gangnon R, Danforth L, Sivakumaran T . Markers of inflammation, oxidative stress, and endothelial dysfunction and the 20-year cumulative incidence of early age-related macular degeneration: the Beaver Dam Eye Study. JAMA Ophthalmol. 2014; 132(4):446-55. PMC: 4076038. DOI: 10.1001/jamaophthalmol.2013.7671. View

10.

Enzbrenner A, Zulliger R, Biber J, Quintela Pousa A, Schafer N, Stucki C . Sodium Iodate-Induced Degeneration Results in Local Complement Changes and Inflammatory Processes in Murine Retina. Int J Mol Sci. 2021; 22(17). PMC: 8431125. DOI: 10.3390/ijms22179218. View

11.

Shen J, Dong A, Hackett S, Bell W, Green W, Campochiaro P . Oxidative damage in age-related macular degeneration. Histol Histopathol. 2007; 22(12):1301-8. DOI: 10.14670/HH-22.1301. View

12.

Zhao Z, Chen Y, Wang J, Sternberg P, Freeman M, Grossniklaus H . Age-related retinopathy in NRF2-deficient mice. PLoS One. 2011; 6(4):e19456. PMC: 3084871. DOI: 10.1371/journal.pone.0019456. View

13.

Mueller-Buehl A, Buehner T, Pfarrer C, Deppe L, Peters L, Dick B . Hypoxic Processes Induce Complement Activation via Classical Pathway in Porcine Neuroretinas. Cells. 2021; 10(12). PMC: 8700265. DOI: 10.3390/cells10123575. View

14.

Seddon J, George S, Rosner B, Rifai N . Progression of age-related macular degeneration: prospective assessment of C-reactive protein, interleukin 6, and other cardiovascular biomarkers. Arch Ophthalmol. 2005; 123(6):774-82. DOI: 10.1001/archopht.123.6.774. View

15.

Carido M, Zhu Y, Postel K, Benkner B, Cimalla P, Karl M . Characterization of a mouse model with complete RPE loss and its use for RPE cell transplantation. Invest Ophthalmol Vis Sci. 2014; 55(8):5431-44. DOI: 10.1167/iovs.14-14325. View

16.

Yang X, Rai U, Chung J, Esumi N . Fine Tuning of an Oxidative Stress Model with Sodium Iodate Revealed Protective Effect of NF-κB Inhibition and Sex-Specific Difference in Susceptibility of the Retinal Pigment Epithelium. Antioxidants (Basel). 2022; 11(1). PMC: 8773095. DOI: 10.3390/antiox11010103. View

17.

Redmond T, Poliakov E, Yu S, Tsai J, Lu Z, Gentleman S . Mutation of key residues of RPE65 abolishes its enzymatic role as isomerohydrolase in the visual cycle. Proc Natl Acad Sci U S A. 2005; 102(38):13658-63. PMC: 1224626. DOI: 10.1073/pnas.0504167102. View

18.

Ferris 3rd F, Wilkinson C, Bird A, Chakravarthy U, Chew E, Csaky K . Clinical classification of age-related macular degeneration. Ophthalmology. 2013; 120(4):844-51. PMC: 11551519. DOI: 10.1016/j.ophtha.2012.10.036. View

19.

Kauppinen A, Niskanen H, Suuronen T, Kinnunen K, Salminen A, Kaarniranta K . Oxidative stress activates NLRP3 inflammasomes in ARPE-19 cells--implications for age-related macular degeneration (AMD). Immunol Lett. 2012; 147(1-2):29-33. DOI: 10.1016/j.imlet.2012.05.005. View

20.

Datta S, Cano M, Ebrahimi K, Wang L, Handa J . The impact of oxidative stress and inflammation on RPE degeneration in non-neovascular AMD. Prog Retin Eye Res. 2017; 60:201-218. PMC: 5600827. DOI: 10.1016/j.preteyeres.2017.03.002. View