A lasered mouse model of retinal degeneration displays progressive outer retinal pathology providing insights into early geographic atrophy

Ibbett, Paul; Goverdhan, Srinivas; Pipi, Elena; Chouhan, Joe K.; Keeling, Eloise; Angus, Elizabeth M.; Scott, Jenny A.; Gatherer, Maureen; Page, Anton; Teeling, Jessica L.; Lotery, Andrew; Ratnayaka, J. Arjuna

doi:10.1038/s41598-019-43906-z

Cited by 18 publications

(16 citation statements)

References 47 publications

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“…In contrast, MC gliosis in mice persists until the last timepoint investigated (Day 14; Figure B, D, E). In line with previous studies [29,30], we showed that our injury model effectively simulates a chronic gliotic response. However, it is unclear how chronic MC reactivity in mammals exacerbates injury response leading to glial scar formation.…”

Section: Discussionsupporting

confidence: 91%

The TGFβ/Notch axis facilitates Müller cell-to-epithelial transition to ultimately form a chronic glial scar

Conedera

Pousa

Mercader

et al. 2020

Preprint

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Background Contrasting with zebrafish, retinal regeneration from Müller cells (MCs) is largely limited in mammals. There, MCs undergo reactive gliosis that consist of a hypertrophic response and ultimately results in vision loss. Transforming growth factor β (TGFβ) is essential for wound healing, including both scar formation and regeneration. However, targeting TGFβ may affect other physiological mechanisms, owing its pleiotropic nature. The regulation of various cellular activities by TGFβ relies on its interaction with other pathways including Notch. Here, we explore the interplay of TGFβ with Notch and how this regulates MC response to injury in zebrafish and mice. Furthermore, we aimed to characterize potential similarities between murine and human MCs during chronic reactive gliosis. Methods Focal damage to photoreceptors was induced with a 532 nm diode laser in TgBAC (gfap:gfap-GFP) zebrafish (ZF) and B6-Tg (Rlbp1-GFP) mice. Transcriptomics, immunofluorescence, and flow cytometry were employed for a comparative analysis of MC response to laser-induced injury between ZF and mouse. The laser-induced injury was paired with pharmacological treatments to inhibit either Notch (DAPT) or TGFβ (Pirfenidone) or TGFβ/Notch interplay (SIS3). To determine if the murine laser-induced injury model translates to the human system, we compared the related MC response to human donors with early retinal degeneration. Results Investigations into injury-induced changes in murine MCs revealed TGFβ/Notch interplay during reactive gliosis. We found that TGFβ1/2 and Notch1/2 interact via Smad3 to reprogram murine MCs towards an epithelial lineage and ultimately to form a glial scar. Similar to what we observed in mice, we confirmed the epithelial phenotype of human Müller cells during gliotic response. Conclusion This study indicates a pivotal role for TGFβ/Notch interplay in tuning MC stemness during injury response and provides novel insights into the remodeling mechanism during retinal degenerative diseases.

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Section: Discussionsupporting

confidence: 91%

The TGFβ/Notch axis facilitates Müller cell-to-epithelial transition to ultimately form a chronic glial scar

Conedera

Pousa

Mercader

et al. 2020

Preprint

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“…The assessment of BEST1 expression, as a marker for viable RPE cells, demonstrated differential expression in the RPE at the affected area of the laser irradiation and progressive RPE cell loss, a hallmark of geography atrophy leading to dry AMD ( Figure 6). Albeit a careful analysis of RPE markers and RPE cells analysis is necessary, the powers above 76 mW with the laser presented here are expected to induce a novel RPE atrophy model [39]. Concomitantly, the results summarized in Figure 7 demonstrate that by applying a heating power of 64-70 mW, 90-100% probability for substantial HSP induction is obtained.…”

Section: Discussionmentioning

confidence: 68%

Induction of Heat Shock Protein 70 in Mouse RPE as an In Vivo Model of Transpupillary Thermal Stimulation

Amirkavei

Pitkänen

Kaikkonen

et al. 2020

IJMS

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The induction of heat shock response in the macula has been proposed as a useful therapeutic strategy for retinal neurodegenerative diseases by promoting proteostasis and enhancing protective chaperone mechanisms. We applied transpupillary 1064 nm long-duration laser heating to the mouse (C57Bl/6J) fundus to examine the heat shock response in vivo. The intensity and spatial distribution of heat shock protein (HSP) 70 expression along with the concomitant probability for damage were measured 24 h after laser irradiation in the mouse retinal pigment epithelium (RPE) as a function of laser power. Our results show that the range of heating powers for producing heat shock response while avoiding damage in the mouse RPE is narrow. At powers of 64 and 70 mW, HSP70 immunostaining indicates 90 and 100% probability for clearly elevated HSP expression while the corresponding probability for damage is 20 and 33%, respectively. Tunel staining identified the apoptotic regions, and the estimated 50% damaging threshold probability for the heating (ED50) was ~72 mW. The staining with Bestrophin1 (BEST1) demonstrated RPE cell atrophy with the most intense powers. Consequently, fundus heating with a long-duration laser provides an approachable method to develop heat shock-based therapies for the RPE of retinal disease model mice.

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“…There are several independent PD [ 75 ] and AMD [ 76 , 77 ] animal models available which emulate the “natural” pathogenesis of each of these human diseases. Thus far, a simple reproducible model that incorporates the features of both AMD+PD in a single model is presently unavailable.…”

Section: Resultsmentioning

confidence: 99%

“…Laser-induced CNV causes retinal degeneration in mouse models [ 77 ]. However, it remains unknown whether PD plays a role in the aggravation of the retinal degeneration in pre-existing AMD animal models.…”

Section: Resultsmentioning

confidence: 99%

“…It is interesting to note that there were large hyperfluorescent progressive vascular lesions persistent at 6 weeks, as visualized by FFA, which insist that chronic exposure to the oral dysbiotic pathogens will exacerbate pathological angiogenesis in the choroidal and retinal area. Other studies have reported the occurrence of hard exudate or drusen-like deposits in laser-induced mice models [ 77 ]. The type, size, and composition are useful biomarkers for analyzing the disease progression.…”

Section: Discussionmentioning

confidence: 99%

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Exacerbation of AMD Phenotype in Lasered CNV Murine Model by Dysbiotic Oral Pathogens

et al. 2021

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Emerging evidence underscores an association between age-related macular degeneration (AMD) and periodontal disease (PD), yet the biological basis of this linkage and the specific role of oral dysbiosis caused by PD in AMD pathophysiology remains unclear. Furthermore, a simple reproducible model that emulates characteristics of both AMD and PD has been lacking. Hence, we established a novel AMD+PD murine model to decipher the potential role of oral infection (ligature-enhanced) with the keystone periodontal pathogen Porphyromonas gingivalis, in the progression of neovasculogenesis in a laser-induced choroidal-neovascularization (Li-CNV) mouse retina. By a combination of fundus photography, optical coherence tomography, and fluorescein angiography, we documented inflammatory drusen-like lesions, reduced retinal thickness, and increased vascular leakage in AMD+PD mice retinae. H&E further confirmed a significant reduction of retinal thickness and subretinal drusen-like deposits. Immunofluorescence microscopy revealed significant induction of choroidal/retinal vasculogenesis in AMD+PD mice. qPCR identified increased expression of oxidative-stress, angiogenesis, pro-inflammatory mediators, whereas antioxidants and anti-inflammatory genes in AMD+PD mice retinae were notably decreased. Through qPCR, we detected Pg and its fimbrial 16s-RrNA gene expression in the AMD+PD mice retinae. To sum-up, this is the first in vivo study signifying a role of periodontal infection in augmentation of AMD phenotype, with the aid of a pioneering AMD+PD murine model established in our laboratory.

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A lasered mouse model of retinal degeneration displays progressive outer retinal pathology providing insights into early geographic atrophy

Cited by 18 publications

References 47 publications

The TGFβ/Notch axis facilitates Müller cell-to-epithelial transition to ultimately form a chronic glial scar

The TGFβ/Notch axis facilitates Müller cell-to-epithelial transition to ultimately form a chronic glial scar

Induction of Heat Shock Protein 70 in Mouse RPE as an In Vivo Model of Transpupillary Thermal Stimulation

Exacerbation of AMD Phenotype in Lasered CNV Murine Model by Dysbiotic Oral Pathogens

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