We executed a genome-wide association scan for age-related macular degeneration (AMD) in 2,157 cases and 1,150 controls. Our results validate AMD susceptibility loci near CFH ( P < 10 ⁻⁷⁵ ), ARMS2 ( P < 10 ⁻⁵⁹ ), C2/CFB ( P < 10 ⁻²⁰ ), C3 ( P < 10 ⁻⁹ ), and CFI ( P < 10 ⁻⁶ ). We compared our top findings with the Tufts/Massachusetts General Hospital genome-wide association study of advanced AMD (821 cases, 1,709 controls) and genotyped 30 promising markers in additional individuals (up to 7,749 cases and 4,625 controls). With these data, we identified a susceptibility locus near TIMP3 (overall P = 1.1 × 10 ⁻¹¹ ), a metalloproteinase involved in degradation of the extracellular matrix and previously implicated in early-onset maculopathy. In addition, our data revealed strong association signals with alleles at two loci ( LIPC , P = 1.3 × 10 ⁻⁷ ; CETP , P = 7.4 × 10 ⁻⁷ ) that were previously associated with high-density lipoprotein cholesterol (HDL-c) levels in blood. Consistent with the hypothesis that HDL metabolism is associated with AMD pathogenesis, we also observed association with AMD of HDL-c—associated alleles near LPL ( P = 3.0 × 10 ⁻³ ) and ABCA1 ( P = 5.6 × 10 ⁻⁴ ). Multilocus analysis including all susceptibility loci showed that 329 of 331 individuals (99%) with the highest-risk genotypes were cases, and 85% of these had advanced AMD. Our studies extend the catalog of AMD associated loci, help identify individuals at high risk of disease, and provide clues about underlying cellular pathways that should eventually lead to new therapies.

Hypoxia-induced VEGF governs both physiological retinal vascular development and pathological retinal neovascularization. In the current paper, the mechanisms of physiological and pathological neovascularization are compared and contrasted. During pathological neovascularization, both the absolute and relative expression levels for VEGF₁₆₄ increased to a greater degree than during physiological neovascularization. Furthermore, extensive leukocyte adhesion was observed at the leading edge of pathological, but not physiological, neovascularization. When a VEGF₁₆₄-specific neutralizing aptamer was administered, it potently suppressed the leukocyte adhesion and pathological neovascularization, whereas it had little or no effect on physiological neovascularization. In parallel experiments, genetically altered VEGF₁₆₄-deficient (VEGF^120/188) mice exhibited no difference in physiological neovascularization when compared with wild-type (VEGF^+/+) controls. In contrast, administration of a VEGFR-1/Fc fusion protein, which blocks all VEGF isoforms, led to significant suppression of both pathological and physiological neovascularization. In addition, the targeted inactivation of monocyte lineage cells with clodronate-liposomes led to the suppression of pathological neovascularization. Conversely, the blockade of T lymphocyte–mediated immune responses with an anti-CD2 antibody exacerbated pathological neovascularization. These data highlight important molecular and cellular differences between physiological and pathological retinal neovascularization. During pathological neovascularization, VEGF₁₆₄ selectively induces inflammation and cellular immunity. These processes provide positive and negative angiogenic regulation, respectively. Together, new therapeutic approaches for selectively targeting pathological, but not physiological, retinal neovascularization are outlined.

Refractive errors, including myopia, are the most frequent eye disorders worldwide and an increasingly common cause of blindness. This genome-wide association meta-analysis in 160,420 participants and replication in 95,505 participants increased the number of established independent signals from 37 to 161 and showed high genetic correlation between Europeans and Asians (>0.78). Expression experiments and comprehensive in silico analyses identified retinal cell physiology and light processing as prominent mechanisms, and also identified functional contributions to refractive-error development in all cell types of the neurosensory retina, retinal pigment epithelium, vascular endothelium and extracellular matrix. Newly identified genes implicate novel mechanisms such as rod-and-cone bipolar synaptic neurotransmission, anterior-segment morphology and angiogenesis. Thirty-one loci resided in or near regions transcribing small RNAs, thus suggesting a role for post-transcriptional regulation. Our results support the notion that refractive errors are caused by a light-dependent retina-to-sclera signaling cascade and delineate potential pathobiological molecular drivers.

Purpose.: To study the choroidal thickness in healthy subjects by swept-source optical coherence tomography (SS-OCT) at longer wavelength. Methods.: The macular area of 31 eyes (31 healthy volunteers) was studied with an SS-OCT prototype system, which uses a tunable laser as a light source operated at 100,000 Hz A scan repetition rate in the 1-μm wavelength region. Three-dimensional volumetric measurement comprised of 512 × 128 A scans was acquired in 0.8 second. From a series of OCT images, a chroidal thickness map of the macular area was created by manual segmentation. To evaluate interoperator reproducibility, the choroidal thickness in each section from 10 subjects was determined independently by two observers. Results.: SS-OCT at the 1-μm wavelength region allowed visualization of the fine structure of the choroid as well as that of the retina. Mean choroidal thickness and volume in the macula area were, respectively, 191.5 ± 74.2 μm and 5.411 ± 2.097 mm³. The mean choroidal thickness of the outer nasal area was significantly thinner than that of all other areas (P < 0.05). The measurements by the two independent observers were significantly identical; the intraclass correlation coefficient in mean choroidal thickness was between 0.945 and 0.980 in each area. The macular choroidal thickness was significantly correlated with axial length after adjustment for age (P < 0.001), and with age after adjustment for axial length (P < 0.001). Conclusions.: SS-OCT system at 1 μm provides macular choroidal thickness maps and allows one to evaluate the choroidal thickness more accurately.

Purpose. The objectives of this study were to characterize the differential potency of two major VEGF isoforms, VEGF₁₂₀ and VEGF₁₆₄, for inducing leukocyte stasis (leukostasis) within the retinal vasculature and blood–retinal barrier (BRB) breakdown and to determine whether endogenous VEGF₁₆₄ mediates retinal leukostasis and BRB breakdown in early and established diabetes. methods. Retinal leukostasis and BRB breakdown were simultaneously quantified by combining concanavalin A lectin (ConA) perfusion labeling with a fluorophotometric dextran leakage assay. CD45 immunohistochemistry was performed to confirm that ConA-stained cells within the vasculature were leukocytes. Retinal leukostasis and BRB breakdown were compared in nondiabetic rats receiving intravitreous injections of VEGF₁₂₀ or VEGF₁₆₄. Retinal intercellular adhesion molecule (ICAM)-1 and VEGF protein levels were studied by Western blot and ELISA, respectively. An anti-VEGF_164(165) aptamer (EYE001) was administered by intravitreous injection to 2-week and 3-month diabetic rats, and the effect on retinal leukostasis and BRB breakdown was quantified. results. Compared with VEGF₁₂₀, VEGF₁₆₄ more potently increased retinal ICAM-1 levels (2.2-fold), leukostasis (1.9-fold), and BRB breakdown (2.1-fold, P < 0.01 for all), despite negligible differences in vitreoretinal VEGF levels at the time of evaluation (P > 0.05). Retinal leukostasis and leakage increased with the duration of diabetes (P < 0.01) and correlated closely (P < 0.01, r = 0.889). The isoform-specific blockade of endogenous VEGF₁₆₄ with EYE001 resulted in a significant suppression of retinal leukostasis and BRB breakdown in both early (72.4% and 82.6%, respectively) and established (48.5% and 55.0%, respectively) diabetes (P < 0.01). conclusions. On an equimolar basis, VEGF₁₆₄ is at least twice as potent as VEGF₁₂₀ at inducing ICAM-1–mediated retinal leukostasis and BRB breakdown in vivo. The inhibition of diabetic retinal leukostasis and BRB breakdown with EYE001 in early and established diabetes indicates that VEGF₁₆₄ is an important isoform in the pathogenesis of early diabetic retinopathy.