OCT Risk Factors for Development of Atrophy in Eyes with Intermediate Age-Related Macular Degeneration.

PURPOSE: To determine the frequency of multiple OCT biomarkers of intermediate age-related macular degeneration (iAMD) and their relationship with the development of complete retinal pigment epithelium and outer retinal atrophy (cRORA) after 2 years. DESIGN: Retrospective cohort study. PARTICIPANTS: This retrospective analysis included 330 eyes of 330 consecutive patients with iAMD in ≥ 1 eye who had 24 months of follow-up data. METHODS: Spectralis OCT volume scans (49 B-scans over 6 × 6 mm, automatic real time = 6, fovea-centered) at baseline were evaluated for the previously described iAMD biomarkers, including a high-central drusen volume (DV; ≥ 0.03 mm3), intraretinal hyper-reflective foci (IHRF), subretinal drusenoid deposits (SDDs), hypo-reflective drusen cores (hDCs), and a thin or thick (multilayered) double-layer sign (DLS). The age-related macular degeneration (AMD) status in the fellow eye was also assessed and classified as normal or early AMD, iAMD, exudative macular neovascularization, or cRORA. MAIN OUTCOME MEASURES: Incidence of cRORA, odds ratio for demographics, and OCT features. RESULTS: At month 24, 16.36% (54/330) of the iAMD eyes developed cRORA. Several baseline features, including high-central DV, IHRF, SDD, hDC, thin DLS, and cRORA in the fellow eye, were associated with a significantly greater risk for development of cRORA at 2 years. The odds ratio, 95% confidence interval, P value, and baseline frequencies of these biomarkers were DV (6.510, 2.467-17.176, P < 0.001, 49.1%), IHRF (12.763, 4.763-34.202, P < 0.001, 38.8%), SDD (2.307, 1.003-5.304, P = 0.049, 34.2%), hDC (3.012, 1.152-7.873, P = 0.024, 13.0%), thin DLS (4.517, 1.555-13.126, P = 0.006, 11.8%), and cRORA in the fellow eye (7.184, 1.938-26.623, P = 0.003, 8.2%). CONCLUSIONS: In addition to the 4 previously reported factors that are present in a significant proportion of iAMD (DV, IHRF, hDC, and SDD), a thin DLS and cRORA in the fellow eye were associated with an increased risk of progression to cRORA over 2 years. These biomarkers may aid in prognostication, risk stratification, and selection of patients for clinical trials. FINANCIAL DISCLOSURE(S): Proprietary or commercial disclosure may be found after the references.

Optical Coherence Tomography Biomarkers for Conversion to Exudative Neovascular Age-related Macular Degeneration.

PURPOSE: To identify optical coherence tomography (OCT) biomarkers, including thin and thick double-layer sign (DLS) for the progression from intermediate AMD (iAMD) to exudative macular neovascularization (MNV) over 24 months. DESIGN: Retrospective cohort study. METHODS: Setting: Retina consultants of Texas. PATIENT POPULATION: 458 eyes of 458 subjects with iAMD in at least 1 eye with 24 months of follow-up data. MAIN OUTCOMES MEASURES: The following biomarkers were assessed at baseline: high central drusen volume (≥0.03 mm3), intraretinal hyper-reflective foci (IHRF), subretinal drusenoid deposits, hyporeflective drusen cores, thick DLS, thin DLS, and central choroidal thickness. A binary logistic regression was computed to investigate the association between baseline OCT covariates and the conversion to exudative MNV within 24 months. In addition, fellow eye status was also included in the model. RESULTS: During follow-up, 18.1% (83 of 458) of eyes with iAMD progressed to exudative MNV. Thick DLS, IHRF, and fellow eye exudative MNV were found to be independent predictors for the development of exudative MNV within 2 years. The baseline frequencies, odds ratios, 95% confidence intervals, and P values for these biomarkers were as follows: thick DLS (9.6%, 4.339, 2.178-8.644; P < .001), IHRF (36.0%, 2.340, 1.396-3.922; P = 0.001), and fellow eye exudative MNV (35.8%, 1.694, 1.012-2.837; P = .045). CONCLUSIONS: Thick DLS, IHRF, and fellow eye exudative MNV were associated with an increased risk of progression from iAMD to exudative MNV. These biomarkers, which are readily identified by the review of OCT volume scans, may aid in risk prognostication for patients and for identifying patients for early intervention trials.

Comparison of Iridocorneal Angle Assessments in Open-Angle Glaucoma and Ocular Hypertension Patients: Anterior Segment Optical Coherence Tomography and Gonioscopy.

Purpose: To quantitatively compare iridocorneal angle assessments using gonioscopy and anterior segment optical coherence tomography (AS-OCT). Patients: US and Chinese patients with open-angle glaucoma (OAG) and/or ocular hypertension (OHT). Methods: Analysis was pooled from 2 multicenter, noninterventional studies conducted in the US and China. Gonioscopy Shaffer grade and an AS-OCT method that approximates the angle width relative to local morphologic variations were compared by measuring the same iridocorneal angles. A third, separate, single-center, noninterventional study was conducted to verify results observed from the pooled analysis. Results: From the pooled studies, a total of 239 eyes were measured using Shaffer grade and AS-OCT. Of these, 6 were Shaffer grade 2, 37 in Shaffer grade 3, and 196 in Shaffer grade 4. There was a trend of increasing Shaffer grade with increasing AS-OCT angle width. Open iridocorneal angles, Shaffer grade ≥3, had a ~98% sensitivity and 88% positive predictive value for identifying AS-OCT angle width ≥300 µm, using the AS-OCT method. To verify these results, a total of 28 right eyes were imaged for the third study. A trend of increasing Shaffer grade with increasing AS-OCT angle width was observed, and angles with Shaffer grade ≤2 had AS-OCT angle width <300 µm. Conclusion: The AS-OCT method can determine the space in the anterior chamber and can potentially identify angles that are the appropriate size for certain glaucoma devices. Information gathered from AS-OCT can provide additional comprehensive and quantitative assessment to gonioscopy.

Subretinal injection in mice to study retinal physiology and disease.

Subretinal injection (SRI) is a widely used technique in retinal research and can be used to deliver nucleic acids, small molecules, macromolecules, viruses, cells or biomaterials such as nanobeads. Here we describe how to undertake SRI of mice. This protocol was adapted from a technique initially described for larger animals. Although SRI is a common procedure in eye research laboratories, there is no published guidance on the best practices for determining what constitutes a 'successful' SRI. Optimal injections are required for reproducibility of the procedure and, when carried out suboptimally, can lead to erroneous conclusions. To address this issue, we propose a standardized protocol for SRI with 'procedure success' defined by follow-up examination of the retina and the retinal pigmented epithelium rather than solely via intraoperative endpoints. This protocol takes 7-14 d to complete, depending on the reagent delivered. We have found, by instituting a standardized training program, that trained ophthalmologists achieve reliable proficiency in this technique after ~350 practice injections. This technique can be used to gain insights into retinal physiology and disease pathogenesis and to test the efficacy of experimental compounds in the retina or retinal pigmented epithelium.

The Learning Curve of Murine Subretinal Injection Among Clinically Trained Ophthalmic Surgeons.

Purpose: Subretinal injection (SRI) in mice is widely used in retinal research, yet the learning curve (LC) of this surgically challenging technique is unknown. Methods: To evaluate the LC for SRI in a murine model, we analyzed training data from three clinically trained ophthalmic surgeons from 2018 to 2020. Successful SRI was defined as either the absence of retinal pigment epithelium (RPE) degeneration after phosphate buffered saline injection or the presence of RPE degeneration after Alu RNA injection. Multivariable survival-time regression models were used to evaluate the association between surgeon experience and success rate, with adjustment for injection agents, and to calculate an approximate case number to achieve a 95% success rate. Cumulative sum (CUSUM) analyses were performed and plotted individually to monitor each surgeon's simultaneous performance. Results: Despite prior microsurgery experience, the combined average success rate of the first 50 cases in mice was only 27%. The predicted SRI success rate did not reach a plateau above 95% until approximately 364 prior cases. Using the 364 training cases as a cutoff point, the predicted probability of success for cases 1 to 364 was 65.38%, and for cases 365 to 455 it was 99.32% (P < 0.0001). CUSUM analysis showed an initial upward slope and then remained within the decision intervals with an acceptable success rate set at 95% in the late stage. Conclusions: This study demonstrates the complexity and substantial LC for successful SRI in mice with high confidence. A systematic training system could improve the reliability and reproducibility of SRI-related experiments and improve the interpretation of experimental results using this technique. Translational Relevance: Our prediction model and monitor system allow objective quantification of technical proficiency in the field of subretinal drug delivery and gene therapy for the first time, to the best of our knowledge.

DDX17 is an essential mediator of sterile NLRC4 inflammasome activation by retrotransposon RNAs.

Detection of microbial products by multiprotein complexes known as inflammasomes is pivotal to host defense against pathogens. Nucleotide-binding domain leucine-rich repeat (NLR) CARD domain containing 4 (NLRC4) forms an inflammasome in response to bacterial products; this requires their detection by NLR family apoptosis inhibitory proteins (NAIPs), with which NLRC4 physically associates. However, the mechanisms underlying sterile NLRC4 inflammasome activation, which is implicated in chronic noninfectious diseases, remain unknown. Here, we report that endogenous short interspersed nuclear element (SINE) RNAs, which promote atrophic macular degeneration (AMD) and systemic lupus erythematosus (SLE), induce NLRC4 inflammasome activation independent of NAIPs. We identify DDX17, a DExD/H box RNA helicase, as the sensor of SINE RNAs that licenses assembly of an inflammasome comprising NLRC4, NLR pyrin domain­containing protein 3, and apoptosis-associated speck-like protein­containing CARD and induces caspase-1 activation and cytokine release. Inhibiting DDX17-mediated NLRC4 inflammasome activation decreased interleukin-18 release in peripheral blood mononuclear cells of patients with SLE and prevented retinal degeneration in an animal model of AMD. Our findings uncover a previously unrecognized noncanonical NLRC4 inflammasome activated by endogenous retrotransposons and provide potential therapeutic targets for SINE RNA­driven diseases.

Identification of fluoxetine as a direct NLRP3 inhibitor to treat atrophic macular degeneration.

The atrophic form of age-related macular degeneration (dry AMD) affects nearly 200 million people worldwide. There is no Food and Drug Administration (FDA)-approved therapy for this disease, which is the leading cause of irreversible blindness among people over 50 y of age. Vision loss in dry AMD results from degeneration of the retinal pigmented epithelium (RPE). RPE cell death is driven in part by accumulation of Alu RNAs, which are noncoding transcripts of a human retrotransposon. Alu RNA induces RPE degeneration by activating the NLRP3-ASC inflammasome. We report that fluoxetine, an FDA-approved drug for treating clinical depression, binds NLRP3 in silico, in vitro, and in vivo and inhibits activation of the NLRP3-ASC inflammasome and inflammatory cytokine release in RPE cells and macrophages, two critical cell types in dry AMD. We also demonstrate that fluoxetine, unlike several other antidepressant drugs, reduces Alu RNA-induced RPE degeneration in mice. Finally, by analyzing two health insurance databases comprising more than 100 million Americans, we report a reduced hazard of developing dry AMD among patients with depression who were treated with fluoxetine. Collectively, these studies identify fluoxetine as a potential drug-repurposing candidate for dry AMD.

Alu complementary DNA is enriched in atrophic macular degeneration and triggers retinal pigmented epithelium toxicity via cytosolic innate immunity.

Long interspersed nuclear element-1 (L1)­mediated reverse transcription (RT) of Alu RNA into cytoplasmic Alu complementary DNA (cDNA) has been implicated in retinal pigmented epithelium (RPE) degeneration. The mechanism of Alu cDNA­induced cytotoxicity and its relevance to human disease are unknown. Here we report that Alu cDNA is highly enriched in the RPE of human eyes with geographic atrophy, an untreatable form of age-related macular degeneration. We demonstrate that the DNA sensor cGAS engages Alu cDNA to induce cytosolic mitochondrial DNA escape, which amplifies cGAS activation, triggering RPE degeneration via the inflammasome. The L1-extinct rice rat was resistant to Alu RNA­induced Alu cDNA synthesis and RPE degeneration, which were enabled upon L1-RT overexpression. Nucleoside RT inhibitors (NRTIs), which inhibit both L1-RT and inflammasome activity, and NRTI derivatives (Kamuvudines) that inhibit inflammasome, but not RT, both block Alu cDNA toxicity, identifying inflammasome activation as the terminal effector of RPE degeneration.

Nucleoside reverse transcriptase inhibitors and Kamuvudines inhibit amyloid-ß induced retinal pigmented epithelium degeneration.

Nonfibrillar amyloid-ß oligomers (AßOs) are a major component of drusen, the sub-retinal pigmented epithelium (RPE) extracellular deposits characteristic of age-related macular degeneration (AMD), a common cause of global blindness. We report that AßOs induce RPE degeneration, a clinical hallmark of geographic atrophy (GA), a vision-threatening late stage of AMD that is currently untreatable. We demonstrate that AßOs induce activation of the NLRP3 inflammasome in the mouse RPE in vivo and that RPE expression of the purinergic ATP receptor P2RX7, an upstream mediator of NLRP3 inflammasome activation, is required for AßO-induced RPE degeneration. Two classes of small molecule inflammasome inhibitors-nucleoside reverse transcriptase inhibitors (NRTIs) and their antiretrovirally inert modified analog Kamuvudines-both inhibit AßOs-induced RPE degeneration. These findings crystallize the importance of P2RX7 and NLRP3 in a disease-relevant model of AMD and identify inflammasome inhibitors as potential treatments for GA.

Cytoplasmic synthesis of endogenous Alu complementary DNA via reverse transcription and implications in age-related macular degeneration.

Alu retroelements propagate via retrotransposition by hijacking long interspersed nuclear element-1 (L1) reverse transcriptase (RT) and endonuclease activities. Reverse transcription of Alu RNA into complementary DNA (cDNA) is presumed to occur exclusively in the nucleus at the genomic integration site. Whether Alu cDNA is synthesized independently of genomic integration is unknown. Alu RNA promotes retinal pigmented epithelium (RPE) death in geographic atrophy, an untreatable type of age-related macular degeneration. We report that Alu RNA-induced RPE degeneration is mediated via cytoplasmic L1-reverse-transcribed Alu cDNA independently of retrotransposition. Alu RNA did not induce cDNA production or RPE degeneration in L1-inhibited animals or human cells. Alu reverse transcription can be initiated in the cytoplasm via self-priming of Alu RNA. In four health insurance databases, use of nucleoside RT inhibitors was associated with reduced risk of developing atrophic macular degeneration (pooled adjusted hazard ratio, 0.616; 95% confidence interval, 0.493-0.770), thus identifying inhibitors of this Alu replication cycle shunt as potential therapies for a major cause of blindness.

A Clinical Metabolite of Azidothymidine Inhibits Experimental Choroidal Neovascularization and Retinal Pigmented Epithelium Degeneration.

Purpose: Azidothymidine (AZT), a nucleoside reverse transcriptase inhibitor, possesses anti-inflammatory and anti-angiogenic activity independent of its ability to inhibit reverse transcriptase. The aim of this study was to evaluate the efficacy of 5'-glucuronyl azidothymidine (GAZT), an antiretrovirally inert hepatic clinical metabolite of AZT, in mouse models of retinal pigment epithelium (RPE) degeneration and choroidal neovascularization (CNV), hallmark features of dry and wet age-related macular degeneration (AMD), respectively. Methods: RPE degeneration was induced in wild-type (WT) C57BL/6J mice by subretinal injection of Alu RNA. RPE degeneration was assessed by fundus photography and confocal microscopy of zonula occludens-1-stained RPE flat mounts. Choroidal neovascularization was induced by laser injury in WT mice, and CNV volume was measured by confocal microscopy. AZT and GAZT were delivered by intravitreous injections. Inflammasome activation was monitored by western blotting for caspase-1 and by ELISA for IL-1ß in Alu RNA-treated bone marrow-derived macrophages (BMDMs). Results: GAZT inhibited Alu RNA-induced RPE degeneration and laser-induced CNV. GAZT also reduced Alu RNA-induced caspase-1 activation and IL-1ß release in BMDMs. Conclusions: GAZT possesses dual anti-inflammatory and anti-angiogenic properties and could be a viable treatment option for both forms of AMD.

Evaluation of retinal vessel quantity within individual retinal structural layers using optical coherence tomography angiography.

PURPOSE: To evaluate retinal vessel quantity within various retinal structural layers using optical coherence tomography angiography (OCTA). METHODS: In this IRB-approved study, 22 normal eyes (from 22 subjects) were imaged using the Spectralis OCT2, with a 15 × 15 degree OCTA scan centered on fovea and two additional 15 × 5 degree OCTA scans, displaced temporally and nasally by 15 degrees along the fovea-Bruch's membrane opening (BMO) axis. Following projection artifact removal (PAR), vessel quantity (i.e., amount of flow signal) within each retinal nuclear and plexiform layer was assessed across the scan and was plotted as a vessel quantity profile over this fovea-BMO axis. Vessel quantity was correlated against the retinal layer thickness at the corresponding locations using the Spearman correlation. RESULTS: For the nerve fiber layer (NFL), the vessel quantity was highest nasally and declined towards the fovea and was near zero temporal to the fovea with or without PAR. For all other retinal layers, the retinal vessel quantities were greatest in the parafoveal retina, peaking approximately 5 degrees from the foveal center. Before PAR, the parafoveal vessel quantity was highest in the inner plexiform layer (IPL). Following PAR, the vessel quantity in the IPL decreased but was relatively unchanged in the other layers. The vessel quantity correlated moderately well with retinal layer thickness (r = 0.432 to 0.511; P < 0.05 among the various layers). CONCLUSIONS: Retinal vessel quantity varies significantly among the various structural layers, with significant regional variability. Projection artifact can significantly impact retinal vessel quantity in the deeper layers, but the effect appears to be most pronounced in the IPL.

Spectral-Domain OCT Analysis of Risk Factors for Macular Atrophy Development in the HARBOR Study for Neovascular Age-Related Macular Degeneration.

PURPOSE: To identify baseline risk factors for macular atrophy (MA) development in HARBOR via a longitudinal assessment of monthly spectral-domain (SD)-OCT scans. Previous analyses of MA in HARBOR examined data from color fundus photography (CFP) and fluorescein angiography (FA). DESIGN: Retrospective, post hoc analysis of SD-OCT images from HARBOR (ClinicalTrials.gov identifier, NCT00891735), a phase 3, multicenter, prospective, randomized, double-blind, active treatment-controlled clinical trial. PARTICIPANTS: Patients (N = 1097) with subfoveal choroidal neovascularization secondary to neovascular age-related macular degeneration (nAMD) treated with intravitreal ranibizumab 0.5 mg monthly (n = 275), 0.5 mg pro re nata (PRN) after 3 loading doses (n = 275), 2.0 mg monthly (n = 274), or 2.0 mg PRN (n = 273). METHODS: Evaluable SD-OCT macular cube scans from patients with 24 months of follow-up (N = 941) were examined monthly from baseline to month 24 by masked reading center-trained graders. Atrophy diagnosis criteria were consistent with those proposed by the Classification of Atrophy Meetings (CAM) group: hypertransmission of light into the choroid, loss of retinal pigment epithelium, and loss of outer retinal layers. Multivariable proportional hazards regression was performed for time to atrophy development. MAIN OUTCOME MEASURES: Risk factors for MA as determined by time to MA development over 24 months of treatment. RESULTS: Baseline risk factors for MA were confirmed from prior analyses that used CFP and FA data: absence of subretinal fluid, presence of intraretinal cysts, presence of Type 3 neovascularization, and presence of atrophy in the fellow eye. This analysis of SD-OCT data identified new baseline risk factors for MA: higher central drusen volume, lower choroidal thickness, presence of nascent atrophy, presence of reticular pseudodrusen, and increased central foveal thickness. Ranibizumab treatment regimen and dose level were not found to be risk factors for MA development. CONCLUSIONS: In this analysis of a major nAMD trial using CAM atrophy criteria, new baseline risk factors for MA development were identified using an SD-OCT dataset. Risk factors for MA development identified by prior analyses were confirmed. Monthly treatment with ranibizumab 0.5 mg was not found to be a risk factor for MA development over 24 months.

Correlation of Quantitative Measurements with Diabetic Disease Severity Using Multiple En Face OCT Angiography Image Averaging.

PURPOSE: To evaluate the effect of averaging en face OCT angiography (OCTA) images on quantitative measurements of the retinal microvasculature and their correlation to diabetic retinopathy (DR) disease severity. DESIGN: Cross-sectional cohort study. PARTICIPANTS: One hundred five eyes (65 patients) comprising 28 eyes from 19 healthy, aged-matched control participants, 14 eyes from 9 diabetics without DR, and 63 eyes from 37 diabetics with varying levels of DR. METHODS: Spectral-domain CIRRUS 5000 (Carl Zeiss Meditec, Dublin, CA) OCTA images with no macular edema or significant motion artifact were acquired 5 times with the 3 × 3-mm scan pattern. En face images of the superficial retinal layer (SRL) and deep retinal layer were registered and averaged. Vessel length density (VLD), perfusion density (PD), and foveal avascular zone (FAZ) parameters were measured on averaged versus single OCTA images. MAIN OUTCOME MEASURES: Univariate and multivariate linear regression correlated quantitative metrics to DR severity and best-corrected visual acuity (BCVA). RESULTS: Eighty-four eyes (55 patients) met the inclusion criteria. Almost uniformly, lower VLD and PD parameters were associated significantly with worse DR severity and BCVA. Multivariate linear regression for DR severity resulted in an R2 value of 0.82 and 0.77 for single and averaged groups, respectively. No variables remained associated significantly with DR severity in multivariate analysis with single images, but in averaged images, increased superior SRL PD significantly predicted worse DR severity (coefficient, 52.7; P = 0.026). Multivariate linear regression for BCVA had an R2 value of 0.42 and 0.47 for single and averaged groups, respectively. Foveal avascular zone size was not associated with DR severity when single OCTA images (P = 0.98) were considered, but was highly associated when using averaged images (coefficient, 6.18; P < 0.001). Foveal avascular zone size was predictive for logarithm of the minimum angle of resolution BCVA with averaged images (0.21; P = 0.004), but not with single images (P = 0.31). CONCLUSIONS: Averaging of en face OCTA images improves the visualization of capillaries, particularly increasing the clarity of the FAZ borders, and therefore improves the correlation of vessel density and FAZ-specific parameters to DR severity and BCVA.

Spectral-Domain OCT-Based Prevalence and Progression of Macular Atrophy in the HARBOR Study for Neovascular Age-Related Macular Degeneration.

PURPOSE: Previous studies of macular atrophy (MA) in HARBOR analyzed color fundus photography and fluorescein angiography image data. This study performed a longitudinal assessment of monthly spectral-domain (SD) OCT scans to determine MA prevalence, incidence, and progression in HARBOR. DESIGN: Post hoc analysis of SD OCT images from HARBOR (ClincalTrials.gov identifier, NCT00891735), a phase 3 multicenter, prospective, randomized, double-blind, active treatment-controlled clinical trial. PARTICIPANTS: Patients (n = 1097) with subfoveal choroidal neovascularization secondary to neovascular age-related macular degeneration (nAMD) treated with ranibizumab 0.5 mg monthly (n = 275), 0.5 mg pro re nata (PRN) after 3 loading doses (n = 275), 2.0 mg monthly (n = 274), or 2.0 mg PRN (n = 273). METHODS: Evaluable SD OCT macular cube scans from patients with 24 months of follow-up (N = 941) were examined by masked reading center-trained graders monthly from baseline to month 24. Atrophy diagnosis criteria were consistent with those proposed by the Classification of Atrophy Meetings (CAM) group: hypertransmission of light into the choroid, retinal pigment epithelium loss, and loss of outer retinal layers. Macular atrophy was considered Definite if all 3 criteria were met and Questionable if 2 were met. Study arms were compared for time to MA detection (log-rank test) and enlargement rates (time × arm interaction test). MAIN OUTCOME MEASURES: Prevalence, incidence, and enlargement rates of MA. RESULTS: At baseline, imbalance in MA rates across ranibizumab arms was evident (0.5 mg monthly, 19.1%; 0.5 mg PRN, 16.1%; 2.0 mg monthly, 10.1%; 2.0 mg PRN, 10.5%). At month 24, new MA development rates in eyes without baseline MA were similar between ranibizumab doses (0.5 mg, 25.9%; 2.0 mg, 25.4%) and treatment regimens (monthly, 26.4%; PRN, 25.0%). No significant differences in enlargement rate of new atrophy area (P = 0.479, square-root transformed) or time to detection of new MA (P = 0.997) were evident among study arms. CONCLUSIONS: In this analysis of a major nAMD trial using CAM atrophy criteria, no differences were observed in incidence or progression rates of new MA among study arms, ranibizumab doses, or treatment regimens. Monthly versus PRN treatment did not influence MA incidence or progression.

OCT Risk Factors for Development of Late Age-Related Macular Degeneration in the Fellow Eyes of Patients Enrolled in the HARBOR Study.

PURPOSE: To evaluate the relationship between OCT features and progression to late age related-macular degeneration (AMD) in the fellow eyes of patients enrolled in the Study of Ranibizumab Administered Monthly or on an As-needed Basis in Patients With Subfoveal Neovascular AMD (HARBOR) (ClinicalTrials.gov identifier, NCT00891735). DESIGN: Post hoc analysis of a phase 3 multicenter, prospective, randomized, double-masked, active treatment-controlled clinical trial. PARTICIPANTS: Evaluable patients (n = 501) with macular neovascularization (MNV) secondary to neovascular AMD and early or intermediate AMD in the fellow eye. METHODS: Volume OCT scans from 501 fellow eyes of 501 patients with MNV were reviewed. Baseline OCT features that were assessed included intraretinal hypereflective foci (IHRF), hyporeflective foci (hRF) within drusenoid lesions (DLs), subretinal drusenoid deposits (SDDs), and drusen volume (DV) of 0.03 mm3 or more. OCT images obtained at months 6, 12, 18, and 24 were graded by masked graders for late AMD (defined as MNV, complete retinal pigment epithelium and photoreceptor atrophy [cRORA], or both). Participant demographic characteristics (age, gender, and smoke exposure) and baseline OCT features were correlated with progression to late AMD. MAIN OUTCOME MEASURES: Incidence of late AMD, hazard ratio (HR) for demographics, and OCT risk factors. RESULTS: At month 24, 33.13% of eyes (166/501) demonstrated late AMD: 20.96% (105/501) demonstrated cRORA, whereas 12.18% (61/501) demonstrated MNV. Baseline demographic factors were not associated significantly with development of late AMD, whereas significant associations were identified for all OCT features. Intraretinal hypereflective foci had an HR of 5.21 (95% confidence interval [CI], 3.29-8.26), hRF within DLs had an HR of 2.42 (95% CI, 1.74-3.38), SDD had an HR of 1.95 (95% CI, 1.34-2.82), and DV of 0.03 mm3 or more had an HR of 1.46 (95% CI, 1.03-2.07). The correlation remained significant when considering only the progression to cRORA and MNV alone, except for DV, which was not associated significantly with progression to MNV. CONCLUSIONS: We confirmed that 4 previously reported OCT risk factors were associated with progression to late AMD in the fellow eyes of patients newly diagnosed with MNV. Although outcomes of more than 2 years were not evaluated, these findings may help to identify high-risk AMD patients.

Repeatability of Fluorescence Lifetime Imaging Ophthalmoscopy in Normal Subjects With Mydriasis.

PURPOSE: We evaluate the repeatability of fluorescence lifetime imaging ophthalmoscopy (FLIO) in normal subjects with mydriasis and explore factors that influence FLIO imaging. METHOD: Thirty-two healthy participants (63 eyes) were enrolled in this prospective study. The Heidelberg Engineering FLIO system uses a 473 nm blue laser light and the emitted fluorescence is detected in two wavelength channels, short and long spectral channels (SSC, LSC). The mean fluorescence lifetime (τm) values were computed for the entire scan area as well as in five regions of interest (ROI, 1 × 1 mm) at the fovea and superior, nasal, inferior, and temporal portions of the macula. Intraclass correlation coefficients (ICC) and coefficients of variation (CV) were used to assess the repeatability. Age, macular thickness, and vascular density also were correlated with τm. RESULTS: The repeatability was good for both channels (ICC, 0.956∼0.995; CV, 9∼16%). The τm for the entire scan was 367.8 ± 58.1 picoseconds (ps) in SSC and 322.5 ± 34.0 ps in LSC. τm was the shortest in the fovea and significantly shorter in the temporal region compared to other regions. τm was positively correlated with age (r = 0.588 for SSC and r = 0.584 for LSC, P = 0.000) and retinal thickness (r = 0.298 for SSC and r = 0.322 for LSC, P = 0.000), and negatively correlated with vascular density (r = -0.112, P = 0.055 for SSC and r = -0.119, P = 0.040 for LSC). CONCLUSION: Repeatable fluorescence lifetime values can be obtained with FLIO, but the lifetimes are affected by age, retinal thickness, vessel density, and macular location. TRANSLATIONAL RELEVANCE: Establishing repeatability of FLIO can introduce fluorescence lifetime imaging technique, which is used in basic science for analysis of excitation and emission wavelength spectrum of fixed and living cells into clinical practice.

A pilot study of fluorescence lifetime imaging ophthalmoscopy in preclinical Alzheimer's disease.

PURPOSE: To investigate fluorescence lifetime imaging ophthalmoscopy (FLIO) findings in preclinical Alzheimer's disease (AD). METHODS: This prospective, observational study enrolled patients with early AD undergoing Alzheimer's biomarker analysis and matched controls. Alzheimer-associated parameters (ß-amyloid [Aß], total tau in cerebrospinal fluid [CSF], Mini-Mental Status Examination [MMSE], etc.), risk factor-associated data (body mass index [BMI], hypertension, lipid profile, etc.), ganglion cell layer plus inner plexiform layer (GCIPL) thickness in structural optical coherence tomography (OCT), OCT angiography data, and FLIO-derived parameters (τm, τ1, τ2, and τ3) in short and long spectral channels (SSC and LSC) were compared and correlated between the two groups. Additional analyses were performed separately within subgroups of phakic and pseudophakic. RESULTS: A total of 28 eyes from 15 subjects (8 control and 7 AD) were included in this analysis. In FLIO parameters, τm in AD group showed longer lifetimes compared to the controls in phakic subjects (593.9 ± 93.3, 454.4 ± 38.6 ps; 475.0 ± 71.6, 394.1 ± 28.2 ps in SSC and LSC of AD and control groups, respectively, p = 0.036 and 0.024). Aß, tau in CSF, and GCIPL thickness correlated with τm in the LSC for phakic subjects (r = -0.611 to 0.562, p < 0.05 for all), but only the GCIPL thickness showed a correlation with FLIO parameters in pseudophakic subjects (r = -0.893 to -0.795, p < 0.001 for all). CONCLUSION: FLIO-derived parameters appear to correlate with Aß, tau levels in the CSF, and GCIPL thickness on OCT in AD patients. If these findings can be validated in future longitudinal studies, FLIO may prove to be useful as a simple, non-invasive diagnostic tool for AD.

Novel and Semiautomated 360-Degree Gonioscopic Anterior Chamber Angle Imaging in Under 60 Seconds.

PURPOSE: Obtaining slit-lamp gonioscopic images requires a high level of expertise and can be time consuming. The images typically are limited to 4 quadrants with limited resolution and can be affected by different lighting conditions. Our goal was to study the ability of the novel imaging system to quickly and easily document the full 360 degrees of the anterior chamber angle in a standardized manner. DESIGN: Cross-sectional cohort study. SUBJECTS: Eighty-four eyes from 50 subjects were included in this study, including 26 eyes from 17 normal participants and 58 eyes from 33 patients diagnosed with glaucoma. METHODS: A newly introduced, commercially available gonioscope (GS-1, NIDEK Co, Gamagori, Japan) was utilized to perform semiautomated imaging of the full 360-degree iridocorneal angle on participants under an institutional review board-approved study. Topical proparacaine was administered to the eye to anesthetize the eye and imaging was completed using the 16-mirror, machine-attached gonioscopic lens coupled with lubricating ophthalmic gel ointment. The first step of the 2-step image acquisition process is analogous to manual gonioscopy, whereas the second step is performed by the instrument in an automated fashion. First, the instrument is manually moved forward by the operator to allow the machine-attached lens to make contact with the patient's corneal surface (time = 30 seconds). Second, the instrument automatically achieves fine focus on the iridocorneal angle and takes 16 sequential high-resolution photographs at multiple different focal planes (time = less than 30 seconds). MAIN OUTCOME MEASURES: Successful qualitative imaging of anterior chamber iridocorneal angle in normal and glaucomatous eyes including visualization of various implanted devices within angle before and after surgery in less than 1 minute per eye. RESULTS: In this pilot study, the full 360 degrees of iridocorneal angle of 84 eyes from 50 normal participants or glaucoma patients were successfully imaged using NIDEK GS-1 in under 60 seconds. All eyes were able to be imaged in 1 sitting, and we were able to image the full spectrum of angle from narrow/closed to fully open. The 360-degree views of the images were able to be displayed in 3 available formats (16-section display, a circular display, or a linear display). Successful imaging was also achieved of surgical devices and postsurgical conditions including CyPass, iStent, XEN, Baerveldt tube shunt, angle recession with iridodialysis, angle neovascularization, pigment dispersion, posttrabeculectomy sclerostomy and surgical iridectomy, and post-laser peripheral iridotomy. CONCLUSION: The newly introduced, semiautomated imaging system was able to document the full 360-degree iridocorneal angle views in normal and glaucoma eyes in less than 1 minute, which may allow longitudinal evaluation in a standardized manner. Furthermore, this system allows the ability to easily evaluate post-angle surgery changes and assess positions of implanted devices in the anterior chamber angle.

Identification and Characterization of Imaging Technique Errors and Artifacts Using Anterior-Segment OCT for Irido-Corneal Angle Evaluations in Glaucoma.

PURPOSE: Anterior-segment OCT (AS-OCT) platforms provide noninvasive and quantifiable evaluations of anterior chamber angle (ACA) anatomy. Although these ACA measurements have been validated with high repeatability and reproducibility, there is a paucity of data previously published regarding the technique errors and artifacts (TEA) that may occur during imaging of the ACA. The purpose of this study was to report on the types and frequency of imaging TEA that are seen in ACA evaluation of a large cohort of patients with open-angle glaucoma. DESIGN: Retrospective, cross-sectional design. PARTICIPANTS: Images were obtained with AS-OCT from 5082 eyes of 2541 patients with glaucoma and were reviewed at the Doheny Image Reading Center (DIRC). METHODS: Images had been captured using AS-OCT devices from 4 different manufacturers (Zeiss Cirrus [Oberkochen, Germany], Heidelberg Spectralis [Heidelberg, Germany], Optovue RTVue [Fremont, CA], or Zeiss Visante). All images were analyzed by 2 independent and certified DIRC image readers for angle opening distance under Schwalbe's line (SL-AOD), and then re-reviewed by a DIRC-certified principal investigator to adjudicate nonconsensus measurements. Images were defined to have TEA when image readers were unable to confidently measure the SL-AOD. MAIN OUTCOME MEASURES: Type and frequency of TEA in the AS-OCT image. RESULTS: Of 5082 eyes reviewed, 208 images (4.09%) were found to have TEA that restricted proper visualization of irido-corneal angle structures and measurements of SL-AOD. The most common TEA was that the image readers were unable to identify Schwalbe's line (74% of TEA images). Other common TEA included poor visualization of the anterior iris surface (15%), misaligned scan location (11%), irregular corneal irregularities (5%), and illumination-related pupillary abnormalities (4%). CONCLUSIONS: This large-scale study presents the types and frequency of TEA found in AS-OCT-derived angle images. More than 95% of the images reviewed were usable for the quantification of irido-corneal angle measurements. Knowing the common types of TEA may help in enhancing the training of both human image readers and in the design of automated algorithms to improve image acquisition and image interpretation parameters. This will become especially useful as AS-OCT becomes more widely used in clinical practice for anterior chamber assessment.