Heterogeneity in the progression of retinal pathologies in mice harboring patient mimicking Impg2 mutations.

Biallelic mutations in interphotoreceptor matrix proteoglycan 2 (IMPG2) in humans cause retinitis pigmentosa (RP) with early macular involvement, albeit the disease progression varies widely due to genetic heterogeneity and IMPG2 mutation type. There are currently no treatments for IMPG2-RP. To aid preclinical studies toward eventual treatments, there is a need to better understand the progression of disease pathology in appropriate animal models. Toward this goal, we developed mouse models with patient mimicking homozygous frameshift (T807Ter) or missense (Y250C) Impg2 mutations, as well as mice with a homozygous frameshift mutation (Q244Ter) designed to completely prevent IMPG2 protein expression, and characterized the trajectory of their retinal pathologies across postnatal development until late adulthood. We found that the Impg2T807Ter/T807Ter and Impg2Q244Ter/Q244Ter mice exhibited early onset gliosis, impaired photoreceptor outer segment maintenance, appearance of subretinal deposits near the optic disc, disruption of the outer retina, and neurosensorial detachment, whereas the Impg2Y250C/Y250C mice exhibited minimal retinal pathology. These results demonstrate the importance of mutation type in disease progression in IMPG2-RP and provide a toolkit and preclinical data for advancing therapeutic approaches.

Reproducibility and staging of 3D human retinal organoids across multiple pluripotent stem cell lines.

Numerous protocols have been described for producing neural retina from human pluripotent stem cells (hPSCs), many of which are based on the culture of 3D organoids. Although nearly all such methods yield at least partial segments of retinal structure with a mature appearance, variabilities exist within and between organoids that can change over a protracted time course of differentiation. Adding to this complexity are potential differences in the composition and configuration of retinal organoids when viewed across multiple differentiations and hPSC lines. In an effort to understand better the current capabilities and limitations of these cultures, we generated retinal organoids from 16 hPSC lines and monitored their appearance and structural organization over time by light microscopy, immunocytochemistry, metabolic imaging and electron microscopy. We also employed optical coherence tomography and 3D imaging techniques to assess and compare whole or broad regions of organoids to avoid selection bias. Results from this study led to the development of a practical staging system to reduce inconsistencies in retinal organoid cultures and increase rigor when utilizing them in developmental studies, disease modeling and transplantation.

Structural and functional findings in patients with moderate diabetic retinopathy.

PURPOSE: To evaluate structural and functional ocular changes in patients with type 2 diabetes mellitus (DM2) and moderate diabetic retinopathy (DR) without apparent diabetic macular edema (DME) assessed by optical coherence tomography (OCT) and microperimetry. METHODS: This was a single-center cross-sectional descriptive study for which 75 healthy controls and 48 DM2 patients with moderate DR were included after applying exclusion criteria (one eye per patient was included). All eyes underwent a complete ophthalmic examination (axial length, macular imaging with swept-source OCT, and MAIA microperimetry). Macular thicknesses, ganglion cell complex (GCC) thicknesses, and central retinal sensitivity were compared between groups, and the relationships between the OCT and microperimetry parameters were evaluated. RESULTS: Macular thickness was similar in both groups (242.17 ± 35.0 in the DM2 group vs 260.64 ± 73.9 in the control group). There was a diminution in the parafoveal area thickness in the DM2 group in the GCC complex. Retinal sensitivity was reduced in all sectors in the DM2 group. The central global value was 24.01 ± 5.7 in the DM2 group and 27.31 ± 2.7 in the control group (p < 0.001). Macular integrity was 80.89 ± 26.4 vs 64.70 ± 28.3 (p < 0.001) and total mean threshold was 23.90 ± 4.9 vs 26.48 ± 2.6 (p < 0.001) in the DM2 and control group, respectively. Moderate correlations were detected between the central sector of MAIA microperimetry and retina total central thickness (- 0.347; p = 0.0035). Age, visual acuity, and hemoglobin A1c levels also correlated with retinal sensitivity. CONCLUSION: Macular GCC thickness and central retinal sensitivity were reduced in patients with moderate DR without DME, suggesting the presence of macular neurodegeneration.

Choroidal Thickness and Volume Modifications Induced by Aerobic Exercise in Healthy Young Adults.

INTRODUCTION: Our aim was to evaluate the changes in choroidal thickness (CT) and volume (CV) following aerobic physical exercise in healthy young adults. METHODS: This study included 72 eyes from healthy volunteers between 22 and 37 years old. Using the International Physical Activity Questionnaire, total physical activity was computed. Measurements using an autorefractometer, ocular biometry, and spectral-domain optical coherence tomography using the Enhanced Depth Imaging protocol were taken. OCT was performed as a baseline measurement and after performing 10 min of dynamic physical exercise (3 and 10 min post-exercise). The choroidal layer was manually segmented, and the CT and CV in different areas from the Early Treatment Diabetic Retinopathy Study grid were obtained. RESULTS: In healthy adults, at 3 min post-exercise, CT was higher in the subfoveal, the 3-mm nasal, and the 6-mm superior areas. Between 3 and 10 min post-exercise, the CT was reduced in all areas, and in some areas, the values were even smaller than the baseline measurements. The CV values showed changes after exercise similar to those of thickness. The total CV recovery after exercise was related to sex and physical activity level. CONCLUSION: Individuals with higher physical activity habits had greater CV at rest than those with lower physical activity levels. During exercise, healthy young people adjust CT and CV. At 3 min post-exercise, CT and CV increase. Women and individuals with greater physical activity levels reduce their total CV more than others during recovery.

CHANGES IN TOTAL AND INNER RETINAL THICKNESSES IN TYPE 1 DIABETES WITH NO RETINOPATHY AFTER 8 YEARS OF FOLLOW-UP.

PURPOSE: To evaluate changes in retinal layer thickness in patients with Type 1 diabetes with no diabetic retinopathy after 8 years of follow-up. METHODS: Ninety Type 1 diabetes and 60 control eyes were studied. Changes in the retinal nerve fiber layer, ganglion cell layer, and inner nuclear layer thicknesses in all Early Treatment Diabetic Retinopathy Study areas were evaluated. RESULTS: The mean ages were 42.93 ± 13.62 and 41.52 ± 13.05 years in the diabetic and control group, respectively. In 2009, total retinal thickness was higher in diabetic patients; differences were statistically significant in all except the nasal areas. In both groups, the mean foveal thickness remained the same during the 8 years. Among diabetic patients, there was a significant reduction in total retinal thickness in all areas excluding the outer temporal one; controls only in the inferior areas. The thickness loss was due to the thinning of the inner retinal layers (inner nuclear layer, ganglion cell layer, and retinal nerve fiber layer). The controls showed a significant diminution in the retinal nerve fiber layer and in the ganglion cell layer areas. The inner nuclear layer showed a diminution in the diabetes mellitus group. CONCLUSION: Before the onset of diabetic retinopathy, Type 1 diabetes patients experience a diminution of their inner retinal layer thicknesses over time, supporting the hypothesis of retinal neurodegeneration.

Measurement method of optical properties of ex vivo biological tissues of rats in the near-infrared range.

An optical fiber-based supercontinuum setup and a custom-made spectrophotometer that can measure spectra from 1100 to 2300 nm, are used to describe attenuation properties from different ex vivo rat tissues. Our method is able to differentiate between scattering and absorption coefficients in biological tissues. Theoretical assumptions combined with experimental measurements demonstrate that, in this infrared range, tissue attenuation and absorption can be accurately measured, and scattering can be described as the difference between both magnitudes. Attenuation, absorption, and scattering spectral coefficients of heart, brain, spleen, retina, and kidney are given by applying these theoretical and experimental methods. Light through these tissues is affected by high scattering, resulting in multiple absorption events, and longer wavelengths should be used to obtain lower attenuation values. It can be observed that the absorption coefficient has a similar behavior in the samples under study, with two main zones of absorption due to the water absorption bands at 1450 and 1950 nm, and with different absolute absorption values depending on the constituents of each tissue. The scattering coefficient can be determined, showing slight differences between retina and brain samples, and among heart, spleen and kidney tissues.

Cellular Characterization of OCT and Outer Retinal Bands Using Specific Immunohistochemistry Markers and Clinical Implications.

PURPOSE: OCT has been a technological breakthrough in the diagnosis, treatment, and follow-up of many ocular diseases, especially retinal and neuro-ophthalmologic pathologic conditions. Until now, several controversies have arisen over the specific cell types that the bands observed in the OCT represent, especially over the 4 outer retinal bands. DESIGN: To correlate the 4 outer hyperreflective bands observed in the OCT with the histologic structures using human retinal sections and immunocytochemistry at the fovea level. PARTICIPANTS: Eyes from human donors. METHODS: Vertical cryosections of human retinas were immunostained with antibodies specific for cones photoreceptors, bipolar cells, mitochondria, Müller cells, and retinal pigment epithelium (RPE) cells and were visualized using confocal microscopy. MAIN OUTCOME MEASURES: Morphological correlation between histology and OCT at the fovea level. RESULTS: Triple immunolabeling allowed distinguishing between cells types and different cell compartments. Immunostaining with guanine nucleotide-binding protein ß 3 (GNB3) and cellular retinaldehyde-binding protein (CRALBP) antibodies showed all retinal layers at the foveola, especially the separation between the outer nuclear layer and the Henle fiber layer. CRALBP and cytochrome C (Cyt C) immunolabeling revealed that hyperreflective bands 1 and 2, observed in the OCT, correspond to the outer limiting membrane and the cone ellipsoids, respectively, separated by the cone myoids. CRALBP, cytochrome C, and GNB3 showed that the RPE interdigitations extend along the entire external segment of the cones, we do not believe them to be the structure responsible for forming the third band. However, the identification of small fragments of cone outer segments within the RPE led us to characterize the third band as the cone phagosomes located in the top of the RPE. Finally, we propose that the fourth band corresponds to the accumulation of mitochondria at the basal portion of the RPE, as identified by cytochrome C immunoreactivity, and that the hyporeflective band between bands 3 and 4 corresponds to the RPE nuclei and melanosomes zone. CONCLUSIONS: This study proposes a new interpretation of the outer retinal bands that leads to a more accurate interpretation of OCT images, providing information about the health of cones and their relationship with the RPE, and could help to form a better understanding of retinal disease diagnosis and prognosis.

Interocular Symmetry of Choroidal Thickness and Volume in Healthy Eyes on Optical Coherence Tomography.

BACKGROUND: We aimed to determine the physiological symmetry, with spectral-domain optical coherence tomography, of choroidal measurements in a healthy population in all the areas defined in the Early Treatment Diabetic Retinopathy Study (ETDRS). METHODS: One hundred and fifty-four eyes of 77 healthy young adults between the ages of 19 and 32 years were enrolled. Differences in choroidal thickness (CT) and volume (CV) between the left and right eyes were calculated. Normal ranges of absolute interocular differences were established as the 95th percentile. RESULTS: The mean ± SD subfoveal CT (SFCT) and total CV values in the right and left eyes were 342.03 ± 77.38 versus 361.64 ± 76.45 µm (correlation coefficient ρ = 0.820; p < 0.001) and 0.27 ± 0.06 versus 0.28 ± 0.06 mm3 (ρ = 0.830; p < 0.001), respectively. Differences in 5 of the 9 areas of the ETDRS map were statistically significant (p < 0.05), but with a strong interocular correlation (ρ > 0.8; p < 0.001). The 95th percentile of interocular tolerance limits for CT in the 1-, 3-, and 6-mm areas were 97, 70, and 57 µm, respectively; the 95th percentile for the volume values were 0.06, 0.51, and 1.73 mm3. CONCLUSIONS: CT and CV are highly correlated between eyes, statistically significant differences between them can be found, and absolute interocular differences may reach 97 µm in SFCT, and 1.73 mm3 in total CV.

Regulation of WNT Signaling by VSX2 During Optic Vesicle Patterning in Human Induced Pluripotent Stem Cells.

Few gene targets of Visual System Homeobox 2 (VSX2) have been identified despite its broad and critical role in the maintenance of neural retina (NR) fate during early retinogenesis. We performed VSX2 ChIP-seq and ChIP-PCR assays on early stage optic vesicle-like structures (OVs) derived from human iPS cells (hiPSCs), which highlighted WNT pathway genes as direct regulatory targets of VSX2. Examination of early NR patterning in hiPSC-OVs from a patient with a functional null mutation in VSX2 revealed mis-expression and upregulation of WNT pathway components and retinal pigmented epithelium (RPE) markers in comparison to control hiPSC-OVs. Furthermore, pharmacological inhibition of WNT signaling rescued the early mutant phenotype, whereas augmentation of WNT signaling in control hiPSC-OVs phenocopied the mutant. These findings reveal an important role for VSX2 as a regulator of WNT signaling and suggest that VSX2 may act to maintain NR identity at the expense of RPE in part by direct repression of WNT pathway constituents. Stem Cells 2016;34:2625-2634.

Loss of MITF expression during human embryonic stem cell differentiation disrupts retinal pigment epithelium development and optic vesicle cell proliferation.

Microphthalmia-associated transcription factor (MITF) is a master regulator of pigmented cell survival and differentiation with direct transcriptional links to cell cycle, apoptosis and pigmentation. In mouse, Mitf is expressed early and uniformly in optic vesicle (OV) cells as they evaginate from the developing neural tube, and null Mitf mutations result in microphthalmia and pigmentation defects. However, homozygous mutations in MITF have not been identified in humans; therefore, little is known about its role in human retinogenesis. We used a human embryonic stem cell (hESC) model that recapitulates numerous aspects of retinal development, including OV specification and formation of retinal pigment epithelium (RPE) and neural retina progenitor cells (NRPCs), to investigate the earliest roles of MITF. During hESC differentiation toward a retinal lineage, a subset of MITF isoforms was expressed in a sequence and tissue distribution similar to that observed in mice. In addition, we found that promoters for the MITF-A, -D and -H isoforms were directly targeted by Visual Systems Homeobox 2 (VSX2), a transcription factor involved in patterning the OV toward a NRPC fate. We then manipulated MITF RNA and protein levels at early developmental stages and observed decreased expression of eye field transcription factors, reduced early OV cell proliferation and disrupted RPE maturation. This work provides a foundation for investigating MITF and other highly complex, multi-purposed transcription factors in a dynamic human developmental model system.

Long time remodeling during retinal degeneration evaluated by optical coherence tomography, immunocytochemistry and fundus autofluorescence.

PURPOSE: To characterize the relationship between fundus autofluorescence (FAF), Optical Coherence Tomography (OCT) and immunohistochemistry (IHC) over the course of chronic retinal degeneration in the P23H rat. METHODS: Homozygous albino P23H rats, Sprague-Dawley (SD) rats as controls and pigmented Long Evans (LE) rats were used. A Spectralis HRA OCT system was used for scanning laser ophthalmoscopy (SLO) imaging OCT and angiography. To determine FAF, fluorescence was excited using diode laser at 488 nm. A fast retina map OCT was performed using the optic nerve as a landmark. IHC was performed to correlate with the findings of OCT and FAF changes. RESULTS: During the course of retinal degeneration, the FAF pattern evolved from some spotting at 2 months old to a mosaic of hyperfluorescent dots in rats 6 months and older. Retinal thicknesses progressively diminished over the course of the disease. At later stages of degeneration, OCT documented changes in the retinal layers, however, IHC better identified the cell loss and remodeling changes. Angiography revealed attenuation of the retinal vascular plexus with time. CONCLUSION: We provide for the first time a detailed long-term analysis of the course of retinal degeneration in P23H rats using a combination of SLO and OCT imaging, angiography, FAF and IHC. Although, the application of noninvasive methods enables longitudinal studies and will decrease the number of animals needed for a study, IHC is still an essential tool to identify retinal changes at the cellular level.

CHOROIDAL THICKNESS AND VOLUME IN A HEALTHY PEDIATRIC POPULATION AND ITS RELATIONSHIP WITH AGE, AXIAL LENGTH, AMETROPIA, AND SEX.

PURPOSE: To evaluate choroidal thickness (CT) and volume in healthy pediatric individuals using enhanced depth imaging spectral domain optical coherence tomography (SD-OCT), as well as its association with age, sex, axial length (AL), and refractive error. METHODS: Ninety-three eyes from 93 healthy pediatric individuals were examined. An Early Treatment Diabetic Retinopathy Study grid was applied to analyze CT and volume map in each of its nine sectors. RESULTS: The mean subfoveal CT and volume were 314.22 ± 55.48 µm and 0.25 ± 0.04 mm, respectively. The nasal CT and volume of both the inner and the outer rings were significantly lower than the temporal area of the same ring and lower than the subfoveal choroidal thickness. A significant negative correlation between the subfoveal CT and AL (r = -0.250, P = 0.015) and a significant positive correlation between the subfoveal CT and refractive error (r = 0.238, P = 0.006) were found. The estimation of the variation in the subfoveal CT in relationship to the AL was -13.55 µm per millimeter. The variation in the subfoveal CT with refractive error was 7.52 µm per diopter. The estimation of the variation in the total choroidal volume related to the AL and ametropia was, respectively, -0.2354 mm per millimeter and 0.1412 mm per diopter. CONCLUSION: Healthy pediatric subjects exhibit choroidal differences in refractive error and AL. In the study population, CT and volume show an increase with age after adjusting for AL.

Induced pluripotent stem cells as custom therapeutics for retinal repair: progress and rationale.

Human pluripotent stem cells have made a remarkable impact on science, technology and medicine by providing a potentially unlimited source of human cells for basic research and clinical applications. In recent years, knowledge gained from the study of human embryonic stem cells and mammalian somatic cell reprogramming has led to the routine production of human induced pluripotent stem cells (hiPSCs) in laboratories worldwide. hiPSCs show promise for use in transplantation, high throughput drug screening, "disease-in-a-dish" modeling, disease gene discovery, and gene therapy testing. This review will focus on the first application, beginning with a discussion of methods for producing retinal lineage cells that are lost in inherited and acquired forms of retinal degenerative disease. The selection of appropriate hiPSC-derived donor cell type(s) for transplantation will be discussed, as will the caveats and prerequisite steps to formulating a clinical Good Manufacturing Practice (cGMP) product for clinical trials.

Electrophysiological findings in long-term type 1 diabetes patients without diabetic retinopathy using different ERG recording systems.

To assess full-field electroretinogram findings in long-term type 1 diabetes patients without diabetic retinopathy. Prospective study including 46 eyes of 23 patients with type 1 diabetes and 46 age-matched healthy eyes evaluated by the RETI-port/scan21 and the portable system RETeval following ISCEV guidelines. The average duration of diabetes was 28.88 ± 8.04 years. In scotopic conditions, using the RETI-port/scan21, diabetic patients showed an increase in b-wave implicit time (IT) (p = 0.017) with the lowest stimuli; a diminished b-wave amplitude (p = 0.005) in the mixed response, an increased IT (p = 0.004) with the high-intensity stimuli and an OP2 increased IT (p = 0.008) and decreased amplitude (p = 0.002). Under photopic conditions, b-wave amplitude was lower (p < 0.001) and 30-Hz flicker response was diminished (p = 0.021). Using the RETeval, in scotopic conditions, diabetic patients showed a reduction in the rod b-wave amplitude (p = 0.009), an increase in a-wave IT with the 280 Td.s stimulus (p = 0.005). OP2 had an increased IT and diminished amplitude (p = 0.003 and p = 0.002 respectively). 16 Td.s flicker showed an increased IT (p = 0.008) and diminished amplitude (p = 0.048). Despite variations in values between both systems, nearly all results displayed positive correlations. Long-term type 1 diabetes patients without diabetic retinopathy exhibit alterations in scotopic conditions, as evidenced by both conventional and portable electroretinogram devices. These findings suggest a modified retinal function, particularly in rod-driven pathways, even in the absence of vascular signs.

Retinal Function in Long-Term Type 1 Diabetes without Retinopathy: Insights from Pattern Electroretinogram and Pattern Visual Evoked Potentials Assessments.

BACKGROUND: To evaluate changes in pattern electroretinogram (pERG) and pattern visual evoked potentials (pVEP) in patients with long-lasting type 1 diabetes without diabetic retinopathy (DR). METHODS: Prospective study involving 92 eyes divided into two groups. The diabetic group included 46 eyes of 23 patients with type 1 diabetes (T1DM); the control group included 23 age-matched healthy subjects. pERG and pVEP were assessed using the RETI-port/scan21 recording software (version 1021.3.0.0). RESULTS: Mean age was 48 ± 9.77 years for the diabetic group and 51.7 ± 4.75 years for the control group. The mean duration of diabetes was 28.88 ± 8.04 years. The mean HbA1c value was 7.29 ± 0.89%. There were no differences in the age or sex distribution. Regarding the pERG, T1DM patients exhibited a significant decrease in the amplitude of the P50 and N95 waves compared to the control group (p = 0.018 and p = 0.035, respectively), with no differences in the peak time of each component. pVEP showed no significant changes in either peak time or amplitude of the different components. CONCLUSIONS: Long-term T1DM patients without DR showed changes in the amplitude of pERG waves with preserved peak times. We did not observe modifications in pVEP. pERG may serve as a subclinical marker of ganglion cell damage in long-term T1DM patients.

Reproducibility and repeatability of Cirrus and Spectralis Fourier-domain optical coherence tomography of healthy and epiretinal membrane eyes.

PURPOSE: To evaluate the accuracy and reproducibility of retinal thickness measurements in healthy and epiretinal membranes (ERM) eyes by Cirrus and Spectralis Fourier-domain optical coherence tomography devices. METHODS: Eighty-seven ERM and 122 healthy subjects underwent 3 macular scans using both optical coherence tomography instruments. Mean thickness measurements in the nine Early Treatment Diabetic Retinopathy Study areas were compared, evaluating the repeatability and the relationship between devices. RESULTS: Macular thickness increase was detected in ERM eyes for all optical coherence tomography parameters (P < 0.001). Mean foveal thickness was 423.5 ± 81.4 and 438.0 ± 54.2 µm for ERM eyes and 267.1 ± 20.2 and 277.5 ± 18.9 µm for healthy eyes using Cirrus and Spectralis, respectively. Macular average thickness in ERM eyes as determined by both optical coherence tomography was correlated (r = 0.812; P < 0.001) but significantly different (P = 0.044). In ERM eyes, measurements showed a mean of the coefficients of variation of 2.95%, 2.2%, and 1.01% using Cirrus, Spectralis, and Spectralis progression feature, respectively. Intraclass correlation coefficients were higher than 0.919 in all cases. CONCLUSION: Reproducibility of both Cirrus and Spectralis optical coherence tomography was high in healthy and ERM eyes. However, considerable differences were found between macular thickness measurements obtained by both devices despite the high correlation between them.

A Reliable Criterion for the Correct Delimitation of the Foveal Avascular Zone in Diabetic Patients.

BACKGROUND: Manual segmentation of the Foveal Avascular Zone (FAZ) has a high level of variability. Research into retinas needs coherent segmentation sets with low variability. METHODS: Retinal optical coherence tomography angiography (OCTA) images from type-1 diabetes mellitus (DM1), type-2 diabetes mellitus (DM2) and healthy patients were included. Superficial (SCP) and deep (DCP) capillary plexus FAZs were manually segmented by different observers. After comparing the results, a new criterion was established to reduce variability in the segmentations. The FAZ area and acircularity were also studied. RESULTS: The new segmentation criterion produces smaller areas (closer to the real FAZ) with lower variability than the different criteria of the explorers in both plexuses for the three groups. This was particularly noticeable for the DM2 group with damaged retinas. The acircularity values were also slightly reduced with the final criterion in all groups. The FAZ areas with lower values showed slightly higher acircularity values. We also have a consistent and coherent set of segmentations with which to continue our research. CONCLUSIONS: Manual segmentations of FAZ are generally carried out with little attention to the consistency of the measurements. A novel criterion for segmenting the FAZ allows segmentations made by different observers to be more similar.

Changes in Inner Retina Thickness and Macular Sensitivity in Patients with Type 2 Diabetes with Moderate Diabetic Retinopathy.

The increase in diabetic retinopathy (DR) prevalence demonstrates the need for the determination of biomarkers for assessing disease development to obtain an early diagnosis and stop its progression. We aimed to analyse total retinal (RT) and inner retinal layer (IRL) thicknesses in type 2 diabetes mellitus (DM2) patients and correlate these results with retinal sensitivity using swept-source OCT (SS-OCT) and microperimetry. For this purpose, a total of 54 DM2 subjects with moderate diabetic retinopathy (DR) with no signs of diabetic macular oedema (DME) and 73 age-matched healthy individuals were assessed using SS-OCT to quantify retinal thickness in the nine macular areas of the ETDRS grid. Retinal sensitivity was measured via microperimetry with a Macular Integrity Assessment Device (MAIA). The mean ages were 64.06 ± 11.98 years for the DM2 group and 60.79 ± 8.62 years for the control group. DM2 patients presented lower visual acuity (p < 0.001) and a thicker RT (260.70 ± 19.22 µm in the control group vs. 271.90 ± 37.61 µm in the DM2 group, p = 0.01). The retinal nerve fibre layer (RNFL) was significantly lower in the outer nasal area (50.38 ± 8.20 µm vs. 45.17 ± 11.25 µm, p = 0.005) in ganglion cells and inner plexiform layers (GCL+) in DM2. A positive correlation between the LDL-C and RNFL and a negative correlation between HDL-C levels and the inner temporal and central RNFL thickness were detected. The central (p = 0.021) and inner nasal (p = 0.01) areas were negatively correlated between the RNFL and MAIA, while GCL++ was positively correlated with the outer inferior (p = 0.015) and outer nasal areas (p = 0.024). Retinal sensitivity and macular RNFL thickness decrease in DM2 patients with moderate DR with no DME, and this study enables an accurate approach to this disease with personalised assessment based on the DR course or stage. Thus, GCL+ and GCL++ thinning may support ganglion cell loss before the RNFL is affected.