Longitudinal changes in outer nuclear layer thickness in soft drusen and reticular pseudodrusen

This is the peer reviewed version of the following article: Ramon, C. , Cardona, G. , Biarnés, M. , Ferraro, L. L. and Monés, J. (2019), Longitudinal changes in outer nuclear layer thickness in soft drusen and reticular pseudodrusen. Clin Exp Optom, 102: 601-610. doi:10.1111/cxo.12894, which has been published in final form at https://doi.org/10.1111/cxo.12894. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.Background: Drusen are seen in the early and intermediate stages of age-related macular degeneration. A retrospective, 2-year observational study at a tertiary centre was designed to assess outer nuclear layer thickness in different types of drusen. Methods: Patients over 50 years with predominant soft drusen or reticular pseudodrusen were included in the study. Fundus photography, infrared, fundus autofluorescence and spectral domain optical coherence tomography were performed at baseline, years 1 and 2. Outer nuclear layer thickness was measured in the nine Early Treatment Diabetic Retinopathy Study subfields, and the rate of thinning was determined using generalized estimating equations models. Results: Data were analysed from 17 eyes with soft drusen and 9 eyes with reticular pseudodrusen. Greater outer nuclear layer thinning was seen overall and in all subfields in reticular pseudodrusen as compared to soft drusen, with statistically significant differences found mostly in superior and nasal subfields of ring 2. The outer nuclear layer was 5-12 µm thinner in eyes with reticular pseudodrusen, and the rate of thinning was greater in eyes with reticular pseudodrusen in the outer superior subfield. Conclusions: In the present sample, outer nuclear layer thickness was consistently lower in patients with reticular pseudodrusen compared with soft drusen, irrespective of subfield location. These structural findings may contribute to explain the functional abnormalities observed in patients with reticular pseudodrusen.Peer ReviewedPostprint (author's final draft

Transplantation of Human Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium in a Swine Model of Geographic Atrophy

Modelo animal; Medicina regenerativa; RetinaModel animal; Medicina regenerativa; RetinaAnimal model; Regenerative medicine; RetinaBackground: The aim of this study was to test the feasibility and safety of subretinal transplantation of human induced pluripotent stem cell (hiPSC)-derived retinal pigment epithelium (RPE) cells into the healthy margins and within areas of degenerative retina in a swine model of geographic atrophy (GA). Methods: Well-delimited selective outer retinal damage was induced by subretinal injection of NaIO3 into one eye in minipigs (n = 10). Thirty days later, a suspension of hiPSC-derived RPE cells expressing green fluorescent protein was injected into the subretinal space, into the healthy margins, and within areas of degenerative retina. In vivo follow-up was performed by multimodal imaging. Post-mortem retinas were analyzed by immunohistochemistry and histology. Results: In vitro differentiated hiPSC-RPE cells showed a typical epithelial morphology, expressed RPE-related genes, and had phagocytic ability. Engrafted hiPSC-RPE cells were detected in 60% of the eyes, forming mature epithelium in healthy retina extending towards the border of the atrophy. Histological analysis revealed RPE interaction with host photoreceptors in the healthy retina. Engrafted cells in the atrophic zone were found in a patchy distribution but failed to form an epithelial-like layer. Conclusions: These results might support the use of hiPSC-RPE cells to treat atrophic GA by providing a housekeeping function to aid the overwhelmed remnant RPE, which might improve its survival and therefore slow down the progression of GA.This work was supported by Spanish Ministry of Science and Innovation (MICINN, RTI2018-095377-B-100), Instituto de Salud Carlos III ISCIII/FEDER (PRB2 PT13/0001/0041; TerCel RD16/0011/0024 and Oftared-RETICS RD16/0008), ERA-NET EuroNanoMed III/ISCIII (AC19/00080), the Catalan Government, AGAUR (2017-SGR-899), CERCA Programme/Generalitat de Catalunya, and by Fundació Carmen i Mª José Godó (Fundació CMJ Godó 2017)

Transplantation of Human Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium in a Swine Model of Geographic Atrophy

Background: The aim of this study was to test the feasibility and safety of subretinal transplantation of human induced pluripotent stem cell (hiPSC)-derived retinal pigment epithelium (RPE) cells into the healthy margins and within areas of degenerative retina in a swine model of geographic atrophy (GA). Methods: Well-delimited selective outer retinal damage was induced by subretinal injection of NaIO3 into one eye in minipigs (n = 10). Thirty days later, a suspension of hiPSC-derived RPE cells expressing green fluorescent protein was injected into the subretinal space, into the healthy margins, and within areas of degenerative retina. In vivo follow-up was performed by multimodal imaging. Post-mortem retinas were analyzed by immunohistochemistry and histology. Results: In vitro differentiated hiPSC-RPE cells showed a typical epithelial morphology, expressed RPE-related genes, and had phagocytic ability. Engrafted hiPSC-RPE cells were detected in 60% of the eyes, forming mature epithelium in healthy retina extending towards the border of the atrophy. Histological analysis revealed RPE interaction with host photoreceptors in the healthy retina. Engrafted cells in the atrophic zone were found in a patchy distribution but failed to form an epithelial-like layer. Conclusions: These results might support the use of hiPSC-RPE cells to treat atrophic GA by providing a housekeeping function to aid the overwhelmed remnant RPE, which might improve its survival and therefore slow down the progression of GA. Keywords: age-related macular degeneration (AMD); geographic atrophy; pig; animal model; stem cells; iPSC; RPE; retina; regenerative medicine; advanced cell therap

Ophthalmology

OBJECTIVE: In the current study we aimed to identify metabolites associated with age-related macular degeneration (AMD) by performing the largest metabolome association analysis in AMD to date. In addition, we aimed to determine the effect of AMD-associated genetic variants on metabolite levels, and aimed to investigate associations between the identified metabolites and activity of the complement system, one of the main AMD-associated disease pathways. DESIGN: Case-control assocation analysis of metabolomics data. SUBJECTS: 2,267 AMD cases and 4,266 controls from five European cohorts. METHODS: Metabolomics was performed using a high-throughput H-NMR metabolomics platform, which allows the quantification of 146 metabolite measurements and 79 derivative values. Metabolome-AMD associations were studied using univariate logistic regression analyses. The effect of 52 AMD-associated genetic variants on the identified metabolites was investigated using linear regression. In addition, associations between the identified metabolites and activity of the complement pathway (defined by the C3d/C3 ratio) were investigated using linear regression. MAIN OUTCOME MEASURES: Metabolites associated with AMD RESULTS: We identified 60 metabolites that were significantly associated with AMD, including increased levels of large and extra-large HDL subclasses and decreased levels of VLDL, amino acids and citrate. Out of 52 AMD-associated genetic variants, seven variants were significantly associated with 34 of the identified metabolites. The strongest associations were identified for genetic variants located in or near genes involved in lipid metabolism (ABCA1, CETP, APOE, LIPC) with metabolites belonging to the large and extra-large HDL subclasses. In addition, 57 out of 60 metabolites were significantly associated with complement activation levels, and these associations were independent of AMD status. Increased large and extra-large HDL levels and decreased VLDL and amino acid levels were associated with increased complement activation. CONCLUSIONS: Lipoprotein levels were associated with AMD-associated genetic variants, while decreased essential amino acids may point to nutritional deficiencies in AMD. We observed strong associations between the vast majority of the AMD-associated metabolites and systemic complement activation levels, independent of AMD status. This may indicate biological interactions between the main AMD disease pathways, and suggests that multiple pathways may need to be targeted simultaneously for successful treatment of AMD

Prevalence of Age-Related Macular Degeneration in Europe: The Past and the Future

Purpose Age-related macular degeneration (AMD) is a frequent, complex disorder in elderly of European ancestry. Risk profiles and treatment options have changed considerably over the years, which may have affected disease prevalence and outcome. We determined the prevalence of early and late AMD in Europe from 1990 to 2013 using the European Eye Epidemiology (E3) consortium, and made projections for the future. Design Meta-analysis of prevalence data. Participants A total of 42 080 individuals 40 years of age and older participating in 14 population-based cohorts from 10 countries in Europe. Methods AMD was diagnosed based on fundus photographs using the Rotterdam Classification. Prevalence of early and late AMD was calculated using random-effects meta-analysis stratified for age, birth cohort, gender, geographic region, and time period of the study. Best-corrected visual acuity (BCVA) was compared between late AMD subtypes; geographic atrophy (GA) and choroidal neovascularization (CNV). Main Outcome Measures Prevalence of early and late AMD, BCVA, and number of AMD cases. Results Prevalence of early AMD increased from 3.5% (95% confidence interval [CI] 2.1%–5.0%) in those aged 55–59 years to 17.6% (95%

Increased fundus autofluorescence, a biomarker of lipofuscin content, as a risk factor for the progression of geographic atrophy secondary to age-related macular degeneration

L’atròfia geogràfica (AG) és la variant avançada de la degeneració macular associada a l’edat seca i es caracteritza per la presència d’àrees d’atròfia de l’epiteli pigmentari de la retina (EPR) que creixen progressivament, amb pèrdua secundària dels fotorreceptors i la coriocapilar adjacents. Actualment és una condició sense tractament. L’acumulació de lipofuscina a l’EPR, visualitzable clínicament mitjançant l’autofluorescència del fons d’ull (AFU) com a zones d’elevada autofluorescència, ha estat vinculada a la progressió de l’AG en alguns estudis. De fet, s’han descrit certs patrons (“fenotips”) en l’AFU basats en la distribució d’hiperautofluorescència que s’han associat amb taxes específiques de progressió de la malaltia. En aquesta Tesi hem fet un estudi clínic prospectiu per simplificar la complexa classificació dels patrons d’AFU i avaluar el paper de la hiperautofluorescència, un biomarcador de la lipofuscina, en la progressió de l’AG.Geographic atrophy (GA) is the advanced form of dry age-related macular degeneration. It is characterized by large areas of retinal pigment epithelium (RPE) atrophy that grow progressively, with concomitant loss of photoreceptors and choriocapillaris. Currently, there is no treatment for this disorder. Lipofuscin build-up within the RPE, which is identifiable by fundus autofluorescence (FAF) as areas of increased autofluorescence, has been linked to GA progression in some studies. Actually, the distribution of hyperautofluorescence on FAF identified some patterns (“phenotypes”), which have been associated with specific rates of disease growth. We conducted a prospective clinical study to simplify the complex classification of FAF patterns and to determine the role of increased FAF, a biomarker of lipofuscin, in the progression of GA

Functional and refractive results after one month of AcrySof toric intraocular lens implantation

Purpose: To describe the functional and refractive results obtained with the implantation of toric intraocular lens (IOL) in a private clinic, and verify and compare these results with other similar studies. Methods: Retrospective evaluation of patients implanted with SN60AT toric IOL. Patients undergoing cataract surgery and corneal astigmatism (CA) higher than 0.75 D were included in the study. Preoperative and postoperative uncorrected visual acuity (UCVA), best corrected visual acuity (BCVA), subjective refraction and preoperative keratometry were recorded and analyzed statistically. Results: The study included 68 eyes (52 patients). Thirty eyes were implanted with SN60T3, 11 with SN60T4 and 28 with SN60T5. Mean BCVA gain was of 1.9 ± 1.67 logMAR lines. Comparing the preoperative BCVA versus postoperative UCVA, the improvement was of 0.89 ± 2 logMAR lines. The postoperative refractive astigmatism was −0.37 ± 0.37 D; 75.5% of the eyes had a refractive astigmatism lower than 0.50 D and 98.6% lower than 1.00 D. The expected cylinder supplied by the manufacturer showed a good agreement with the postoperative subjective results (−0.03 ± 0.47 D). Conclusions: The implantation of SN60T toric IOL in patients with CA higher than 0.75 D is a safe, predictable and effective way of reducing refractive astigmatism in patients undergoing cataract surgery

Early Detection of Incipient Retinal Pigment Epithelium Atrophy Overlying Drusen with Fundus Autofluorescence vs. Spectral Domain Optical Coherence Tomography

Purpose. This study aims to find out which tool, fundus autofluorescence (FAF) or spectral domain optical coherence tomography (SD-OCT), is more sensitive in detecting retinal pigment epithelium (RPE) demise overlying drusen and can, therefore, help predict geographic atrophy (GA) appearance in Age-Related Macular Degeneration (AMD). Methods. A single-site, retrospective, observational, longitudinal study was conducted. Patients with intermediate AMD (iAMD) (large (>125 μm) or intermediate (63–125 μm) drusen with hyper/hypopigmentation) with a minimum follow-up of 18 months were included. Drusen with overlying incipient RPE atrophy were identified on SD-OCT defined as choroidal hypertransmission or nascent geographic atrophy (nGA). These selected drusen were, then, traced backwards in time to determine if incipient RPE atrophy overlying drusen was observed on FAF (well-demarcated region of absence of autofluorescence) before, simultaneously, or after having detected the first signs of incipient RPE atrophy on SD-OCT. The number of drusen in which signs of incipient RPE atrophy was detected earlier using FAF or SD-OCT was compared. The time elapsed from the identification with the more sensitive method to the other was recorded and analyzed. Results. One hundred and thirty-three drusen in 22 eyes of 22 patients were included. Of these, 112 (84.2%) drusen showed choroidal hypertransmission and 21(15.8%) nGA. Early signs of atrophy overlying drusen were found simultaneously on SD-OCT and FAF in 52 cases (39.1%, 95% CI 30.8–47.9%), earliest on FAF in 51 (38.3%, 95% CI 30.0–47.2%) and first on SD-OCT in 30 (22.6%, 95% CI 15.8–30.6%; p<0.05). Statistically significant differences were found between both techniques (p=0.005), with FAF detecting it earlier than SD-OCT. When RPE atrophy was found first on FAF, the median time to diagnosis with SD-OCT was 6.6 months (95% CI 5.5 to 8.6), while if detection occurred earlier on SD-OCT, the median time until identification with FAF was 12.6 months (95% CI 6.0 to 23.4; p=0.0003). Conclusions. In iAMD cases in which early atrophy overlying drusen is not detected simultaneously in FAF and SD-OCT, FAF was significantly more sensitive. Nevertheless, a multimodal approach is recommended and required to evaluate these patients

Longitudinal changes in outer nuclear layer thickness in soft drusen and reticular pseudodrusen

This is the peer reviewed version of the following article: Ramon, C. , Cardona, G. , Biarnés, M. , Ferraro, L. L. and Monés, J. (2019), Longitudinal changes in outer nuclear layer thickness in soft drusen and reticular pseudodrusen. Clin Exp Optom, 102: 601-610. doi:10.1111/cxo.12894, which has been published in final form at https://doi.org/10.1111/cxo.12894. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.Background: Drusen are seen in the early and intermediate stages of age-related macular degeneration. A retrospective, 2-year observational study at a tertiary centre was designed to assess outer nuclear layer thickness in different types of drusen. Methods: Patients over 50 years with predominant soft drusen or reticular pseudodrusen were included in the study. Fundus photography, infrared, fundus autofluorescence and spectral domain optical coherence tomography were performed at baseline, years 1 and 2. Outer nuclear layer thickness was measured in the nine Early Treatment Diabetic Retinopathy Study subfields, and the rate of thinning was determined using generalized estimating equations models. Results: Data were analysed from 17 eyes with soft drusen and 9 eyes with reticular pseudodrusen. Greater outer nuclear layer thinning was seen overall and in all subfields in reticular pseudodrusen as compared to soft drusen, with statistically significant differences found mostly in superior and nasal subfields of ring 2. The outer nuclear layer was 5-12 µm thinner in eyes with reticular pseudodrusen, and the rate of thinning was greater in eyes with reticular pseudodrusen in the outer superior subfield. Conclusions: In the present sample, outer nuclear layer thickness was consistently lower in patients with reticular pseudodrusen compared with soft drusen, irrespective of subfield location. These structural findings may contribute to explain the functional abnormalities observed in patients with reticular pseudodrusen.Peer Reviewe