Shin Yoneya

Phenotype and Genotype Characteristics of Age-related Macular Degeneration in a Japanese Population

PURPOSE To describe phenotype and genotype characteristics of age-related macular degeneration (AMD) in Japanese patients. DESIGN A case-control study. PARTICIPANTS A total of 550 case-control samples composed of 408 consecutive AMD cases and 142 controls. METHODS Clinical information assessing age, gender, affected eyes, fundus features, and fluorescein/indocyanine green angiograms were systematically evaluated. Four single nucleotide polymorphisms (SNPs; rs800292, rs1061170, rs1410996, rs2274700) in the complement factor H (CFH) gene, 1 SNP (rs11200638) in the high-temperature requirement factor A1 (HTRA1) gene, 3 SNPs (rs699947, rs1570360, rs2010963) in the vascular endothelial growth factor (VEGF) gene, and 4 SNPs (rs12150053, rs12948385, rs9913583, rs1136287) in the pigment epithelium-derived factor (PEDF) gene were assessed using TaqMan technology. MAIN OUTCOME MEASURES The clinical phenotype information and genotypes of CFH, HTRA1, VEGF, and PEDF polymorphisms. RESULTS Of Japanese patients with neovascular AMD (nAMD), 219 (58.7%) had typical nAMD and 154 (41.3%) had polypoidal choroidal vasculopathy (PCV). The frequency of bilateral exudative involvement was similar between typical nAMD (15.5%) and PCV (13.6%) (P = 0.613). Significant soft drusen were observed in the fellow eyes of 88 (47.6%) of 185 patients with unilateral typical nAMD and in 25 (18.8%) of 133 patients with unilateral PCV (P = 1.24x10(-7)). A serous pigment epithelium detachment was seen in 55 (25.1%) of 219 patients with typical nAMD and in 64 (41.6%) of 154 patients with PCV. A significant association was noted in CFH-rs800292, CFH-rs1410996, CFH-rs2274700, and HTRA1-rs11200638 with AMD development (P = 2.36x10(-5), 7.18x10(-5), 7.18x10(-5), 2.70x10(-7), respectively; population attributable risk = 57.3%, 57.8%, 57.8%, and 58.9%, respectively). We estimated the highest-risk group to have an approximately 70-fold greater risk of nAMD compared with the lowest-risk group when analyzing a combination of 4 SNPs in the CFH and HTRA1 genes. CONCLUSIONS The Japanese AMD phenotype is characterized by a higher frequency of PCV, male predominance, and lower frequency of bilateral presentation compared with Caucasian AMD. Genotype analyses demonstrate a significant population attributable risk for SNPs in the CFH and HTRA1 genes and demonstrate joint effects for both genes. Gene variants in both CFH and HTRA1 contribute significantly to the AMD phenotype in a Japanese population.

Comparison of epiretinal membranes of differing pathogenesis using optical coherence tomography.

Purpose To compare optical coherence tomography (OCT) images of idiopathic epiretinal membranes (ERMs) with those of secondary ERMs. Methods OCT was performed on 70 eyes of 63 consecutive patients with biomicroscopic evidence of ERMs and 23 eyes of 23 healthy volunteers without ERMs. OCT findings were correlated with the clinical pathogenesis of the ERM. Results Evaluation by OCT established that 48 of 70 ERMs were globally adherent to the retina and that 22 of 70 ERMs were focally adherent to the retina. When correlated to clinical pathogenesis, 20% of idiopathic membranes and 52% of secondary membranes were focally attached to the retina. There was a significant difference in the pattern of membrane attachment to the retina in the two pathogenic groups (P = 0.007). Eight of nine eyes with macular pseudoholes were associated with globally adherent membranes. Conclusion Secondary ERMs are more likely to be characterized by focal retinal adhesion than are primary ERMs. Primary ERMs tend to be globally adherent. This finding may contribute to understanding the underlying mechanisms of ERM formation in different clinical settings.

Intraocular gene transfer of pigment epithelium-derived factor rescues photoreceptors from light-induced cell death

In this study, we investigated whether intraocular gene transfer of pigment epithelium‐derived factor (PEDF) ameliorates the extent of light‐induced photoreceptor cell death. Lewis rats received intravitreous injection of 3 × 109 particles of adenoviral vector expressing PEDF (AdPEDF.11) in one eye and 3 × 109 particles of empty adenoviral vector (AdNull.11) in the contralateral eye. The rats were then dark‐adapted for 3 days after which they were continuously exposed to fluorescent light (2,500 lux) for 0, 6, 24, 96, and 168 h. Both eyes were then enucleated and processed for morphometric analysis. Cell death in the retina was examined using TUNEL staining with a propidium iodide counterstain. The photoreceptor cell counts in each of the three groups were significantly different (P < 0.001). Eyes that received intravitreous injection of AdNull.11 or no injection showed a greater number of pyknotic photoreceptor cells and a reduced photoreceptor cell density as compared to eyes treated with intravitreous AdPEDF.11 injection. AdNull.11 treated eyes showed a lesser but still significant protection of photoreceptor cells when compared to untreated eyes. Fewer TUNEL‐positive photoreceptor cells were present in AdPEDF.11 treated eyes than in AdNull.11 treated or untreated eyes (P = 0.004). The amplitudes of the ERG a‐wave, b‐wave, and oscillatory potentials (OPs) were increased significantly by treatment (P < 0.05). These data suggest that adenovirus vector‐mediated intraocular expression of PEDF significantly increases photoreceptor cell survival following excessive light exposure. Neuroprotection may result from inhibition of light‐induced apoptotic processes. This study provides proof of concept for a gene transfer approach to modulating retinal cell death resulting from photo‐oxidative damage and supports the hypothesis that gene transfer of PEDF is broadly applicable to modulating apoptosis in the retina.

Complement Factor H and High-Temperature Requirement A-1 Genotypes and Treatment Response of Age-related Macular Degeneration

PURPOSE To determine whether there is an association between complement factor H (CFH), high-temperature requirement A-1 (HTRA1), vascular endothelial growth factor (VEGF), and pigment epithelium-derived factor (PEDF) genotypes and response to treatment with photodynamic therapy (PDT) for age-related macular degeneration (AMD) in a Japanese population. DESIGN Prospective, case-control study. PARTICIPANTS One hundred ten patients with exudative AMD treated by verteporfin PDT were recruited prospectively at the Department of Ophthalmology, Saitama Medical University Hospital, Saitama, Japan. METHODS The patients were genotyped for 4 single nucleotide polymorphisms (SNPs; rs800292, rs1061170, rs1410996, rs2274700) in the CFH gene, a rs11200638-SNP in the HTRA1 gene, 3 SNPs (rs699947, rs1570360, rs2010963) in the VEGF gene, and 4 SNPs (rs12150053, rs12948385, rs9913583, rs1136287) in the PEDF gene using a TaqMan assay. MAIN OUTCOME MEASURES The treatment outcomes and genotypes of CFH, HTRA1, VEGF, and PEDF polymorphisms. RESULTS Best-corrected visual acuity 1 year after PDT was significantly increased in patients with the HTRA1-rs11200638 GG genotype as compared with patients with the GA or AA genotypes (P = 2.9 × 10⁻², 7.0 × 10⁻⁴, respectively). The rate of recurrence in the 12-month period after PDT was also associated with HTRA1-rs11200638 genotype (P = 3.12 × 10⁻²). Patients with the AA genotype of HTRA1-rs11200638 had an approximately 6-fold greater risk of the recurrence than patients with the GG genotype (P = 5.58 × 10⁻³). Significant differences were demonstrated in the mean time interval from the initial treatment to the time of recurrence for the genotypes of CFH-rs1410996/-rs2274700 (P = 8.50 × 10⁻³). CONCLUSIONS The HTRA1-rs11200638 and CFH-rs1410996/-rs2274700 variants were associated with response to PDT in this study population. These variants may be used for genetic biomarkers to estimate visual outcomes and recurrences in the response to PDT with significant predictive power.

Association of the HTRA1 gene variant with age-related macular degeneration in the Japanese population

AbstractThe purpose of this investigation was to determine whether the high-temperature requirement A-1 (HTRA1) gene polymorphism is associated with age-related macular degeneration (AMD) in native, unrelated Japanese patients. A total of 123 patients with AMD and 133 control subjects without AMD were recruited for this study. The single-nucleotide polymorphism (SNP) rs11200638 in the HTRA1 gene was assessed using a TaqMan assay. The risk A allele frequencies in the AMD cases and control patients were 0.577 and 0.380, respectively, and were associated with a significant risk of developing AMD (p=7.75×10−6). The results were more significant in subtype analyses with wet AMD (p=5.96×10−7). We conclude that the rs11200638 variant in the HTRA1 gene is strongly associated with AMD in the Japanese population. This result supports the hypothesis that the HTRA1 gene may increase susceptibility to AMD development and can participate in a potential new molecular pathway for AMD pathogenesis by extending this association across diverse ethnicities.

Retinal oxygen saturation levels in patients with central retinal vein occlusion

PURPOSE To validate the efficacy of Fourier transform-based spectral retinal imaging (SRI) in quantifying retinal ischemia. DESIGN Prospective, observational case series. PATIENTS AND METHODS Eleven eyes of 10 patients with central retinal vein occlusion (CRVO) and 4 fellow unaffected eyes of selected patients were examined by both fluorescein angiography and SRI. The fluorescein angiograms were correlated with oxygen saturation maps that were calculated from the SRI. RESULTS Oxygen saturation levels in the fundus were shown as color grading in 35 degrees fundus images processed by SRI. This grading correlated well with the severity of CRVO, as estimated by fluorescein angiography. CONCLUSIONS The severity of circulatory disturbances in CRVO may be graded by applying SRI.

Aging changes of the choroidal dye filling pattern in indocyanine green angiography of normal subjects.

Purpose To study the aging changes in the choroid of healthy volunteers with indocyanine green (ICG) angiography. Methods Video ICG angiography with adjunctive computer-assisted image analysis was performed on 35 eyes of 30 healthy volunteers (age range, 21–81 years; mean ± standard deviation, 50.5 ± 16.2 years) to observe the aging changes of the choroid. Results In patients in the second and third decades of life, the subfoveal choroidal arterioles fluoresced initially with subsequent rapid filling of the feeding arterioles and choriocapillaris. The watershed zone was clearly observed. In patients older than age 50, the choroidal vasculature filled more slowly. Eventually, the margin of the watershed zone became blurred. The quantitative analysis showed that the number of choroidal arterioles and the fluorescent intensity in the macular region were reduced with age (P < 0.005). In the early venous phase, hypofluorescent patches seen in all ages increased in size and number and remained with aging. The mean fluorescence intensity was not correlated statistically with age. Conclusions The features of normal aging patterns of the choroid that we investigated are essential to the interpretation of ICG angiography and may help in understanding the physiologic and pathologic conditions of the choroidal circulations and the choroid and retina themselves.

Associations of cigarette smoking but not serum fatty acids with age-related macular degeneration in a Japanese population.

PURPOSE To assess modifiable environmental risk factors and protective factors for age-related macular degeneration (AMD) in a native Japanese population. DESIGN A case-control study. PARTICIPANTS We included 422 case-control samples composed of 279 consecutive AMD cases and 143 controls. METHODS Information regarding systemic conditions and lifestyle were documented in each subject by standardized questionnaire including age, gender, smoking history, body mass index (BMI), and history of cardiovascular disease, hypertension, and diabetes. Serum fatty acids profiles were analyzed by gas chromatography performed on blood samples taken from each study participant. Logistic regression and multiple comparison analyses were utilized in this study. MAIN OUTCOME MEASURES Population-specific information assessing systemic conditions, lifestyle, and serum fatty acid profiles. RESULTS Among environmental factors analyzed cigarette smoking showed the most significant association with development of all AMD (P<0.00001; odds ratio [OR], 4.06; 95% confidence interval [CI], 2.22-7.43), typical neovascular AMD (P<0.0001, OR, 4.59; 95% CI, 2.29-9.18), and polypoidal choroidal vasculopathy (P<0.001; OR, 4.87; 95% CI, 1.96-12.1). Hypertension and BMI showed a mild association with AMD. Although male prevalence was significantly higher in all case groups than in controls with conventional Scheffe correction, there was no association of gender with AMD development when logistic regression analysis was used to adjust for cigarette smoking. There was no difference in fatty acid profiles, except for a mild association of eicosapentaenoic acid concentration in the all AMD group. CONCLUSIONS In the Japanese population studied, cigarette smoking influenced the risk of AMD but fractionated serum fatty acid levels did not. Although prior reports indicate a male predominance in Japanese patients with AMD, this study demonstrates that cigarette smoking accounts for this confounding bias. In addition, our population-specific data do not demonstrate significant differences in serum fatty acid composition, including ω-3 and ω-6 long chain polyunsaturated fatty acids, in Japanese patients with and without AMD. These results are consistent with the high proportion of smokers in aged Japanese men and the high fish oil intake in this population. FINANCIAL DISCLOSURE(S) The authors have no proprietary or commercial interest in any of the materials discussed in this article.

Problems with and pitfalls of photodynamic therapy.

OBJECTIVE To delineate the various factors that may influence the outcome of photodynamic therapy of the retina and choroid. DESIGN Experimental animal study. ANIMALS Pigmented and nonpigmented rabbits; rhesus monkeys. INTERVENTION The hydrophilic photosensitizer, mono-L-aspartyl chlorin e6, which is maximally activated at 664 nm, was studied after intravenous injection into pigmented and nonpigmented rabbits and rhesus monkeys. Laser light was supplied by a red diode laser coupled to a modified slit-lamp biomicroscope and delivered to the ocular fundus after passing through a standard fundus contact lens. Standard photodynamic parameters were used. The effects of fundus pigmentation, intraocular pressure, spot focus and defocus, region of fundus treated, equivalent fluence, and retreatment were observed in the different animal species. MAIN OUTCOME MEASURES Slit-lamp biomicroscopy, fluorescein angiography, light and transmission electron microscopy. RESULTS Fundus pigmentation appeared to be a factor only at the lowest fluence level tested, where only 4 of 12 lesions attempted in pigmented fundi were noted on fluorescein angiography, compared with 12 of 12 lesions in albino rabbits. At normal intraocular pressures and a given fluence, 10 of 10 lesions were fully manifested on fluorescein angiography, compared with 4 of 10 at 30 mmHg and 0 of 10 at pressures sufficient to blanch the optic nerve (>60 mmHg). For laser spots either focused or defocused, there were 6 of 6 lesions that were fully manifested on fluorescein angiography for each of the parameters. Lesions treated in the fovea resulted in larger spots on fluorescein angiography. The fluence of 5 mW for 10 seconds resulted in a larger lesion on angiography than the equivalent fluence of 10 mW for 5 seconds. Areas of retreatment in rabbits demonstrated more thinning of the neurosensory retina and loss of photoreceptor outer segments and nuclei than corresponding areas receiving one treatment. CONCLUSIONS Photodynamic therapy results varied, depending on intraocular pressure, region of fundus treated, ocular pigmentation, and the total time of exposure to the photosensitizer. Retreatment resulted in progressive thinning of the neurosensory retina with loss of photoreceptor outer segments and nuclei in the rabbit eye.

Angiographic and histologic effects of fundus photodynamic therapy with a hydrophilic sensitizer (mono-l-aspartyl chlorin e6)

PURPOSE To demonstrate the efficacy of the photosensitizer mono-L-aspartyl chlorin e6 (NPe6) in closing choroidal vessels at low energy levels, that tissue uptake and clearance are rapid, and that low concentrations of drug are needed to achieve clinical effects. DESIGN Experimental animal study. ANIMALS Pigmented rabbits and Japanese monkeys were used in this study. METHODS Using a modified 664-nm diode laser, the fundi of pigmented rabbits and Japanese monkeys were irradiated after intravenous administration of NPe6 (2-100 mg/kg). Time from injection to irradiation varied from 5 to 15 minutes, and duration of exposure varied from 1 to 10 seconds. Power output at the corneal surface was either 3.6 or 5.9 mW. Animals were examined by indirect ophthalmoscopy and fluorescein angiography at 2 hours and 7 days after treatment. After enucleation 7 days after treatment, specimens were prepared for light and electron microscopy. MAIN OUTCOME MEASURES Angiographic evidence of occlusion and histopathologic evidence of retinal damage. RESULTS Both clinical and histopathologic examination demonstrated effects on the choroidal vasculature and the retinal pigment epithelium, including necrosis of endothelial cells and occlusion in choroidal vessels, particularly within the choriocapillaris, at low energy levels. Overlying neurosensory retina was minimally affected. Fluorescein angiography of lesions treated with 2 mg/kg and laser fluence of 2.3 to 7.5 J/cm2 showed a normal appearance 2 hours after treatment, which changed to early hypofluorescent and later hyperfluorescent lesions 7 days after treatment. In contrast, those animals receiving the 10-mg/kg dose and laser fluence of 0.46 to 0.75 J/cm2 showed marked hypofluorescence of choroidal lesions and occlusion of retinal vessels 7 days after treatment. CONCLUSIONS Effective occlusion of normal choroidal vessels was achieved at 2 mg/kg using 2.3 to 7.5 J/cm2 or at 10 mg/kg using 0.46 to 0.75 J/cm2 with minimal injury to overlying neurosensory retina.