Profile of Lipoprotein Subclasses in Chinese Primary Open-Angle Glaucoma Patients.

To investigate the plasma lipoprotein subclasses in patients with primary open-angle glaucoma (POAG), a total of 20 Chinese POAG patients on intraocular pressure (IOP)-lowering treatment and 20 age-matched control subjects were recruited. Based on the levels of total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C), the study subjects were divided into elevated- and normal-level subgroups. The plasma lipoprotein, lipoprotein subclasses, and oxidized LDL (oxLDL) levels were quantitatively measured. The discrimination potential of the lipoproteins was evaluated using the area under the receiver operating characteristic curve (AUC), and their correlation with clinical parameters was also evaluated. Compared to the control subjects with elevated TC and/or LDL-C levels, the levels of TC, LDL-C, non-high-density lipoprotein cholesterol (non-HDL), LDL subclass LDL3 and small dense LDL (sdLDL), and oxLDL were significantly higher in POAG patients with elevated TC and/or LDL-C levels. No differences in any lipoproteins or the subclasses were found between the POAG patients and control subjects with normal TC and LDL-C levels. Moderate-to-good performance of TC, LDL-C, non-HDL, LDL3, sdLDL, and oxLDL was found in discriminating between the POAG patients and control subjects with elevated TC and/or LDL-C levels (AUC: 0.710-0.950). Significant negative correlations between LDL3 and sdLDL with retinal nerve fiber layer (RNFL) thickness in the superior quadrant and between LDL3 and average RNFL thickness were observed in POAG patients with elevated TC and/or LDL-C levels. This study revealed a significant elevation of plasma lipoproteins, especially the LDL subclasses, in POAG patients with elevated TC and/or LDL-C levels, providing insights on monitoring specific lipoproteins in POAG patients with elevated TC and/or LDL-C.

Primary open-angle glaucoma risk prediction with ABCA1 and LOC102723944 variants and their genotype-phenotype correlations in southern Chinese population.

Glaucoma is a leading cause of irreversible visual impairment and blindness worldwide. Previous genome-wide association studies have identified caveolin-1 (CAV1), ATP-binding cassette A1 (ABCA1), and forkhead box C1 (FOXC1) loci associated with primary open angle glaucoma (POAG), a major subtype of glaucoma. This study aimed to fine map the association pattern of FOXC1 locus with POAG and determine the correlations of FOXC1, ABCA1, and CAV1 variants with ocular and lipidemic parameters in southern Chinese population. In total, 1291 unrelated Han Chinese subjects were recruited, including 301 high-tension glaucoma (HTG), 126 normal-tension glaucoma (NTG), and 864 control subjects. Twelve variants in FOXC1 locus, and two variants in ABCA1 and CAV1 genes, were genotyped by TaqMan assays. Genetic risk score and genotype-phenotype correlation analyses were conducted. In the FOXC1 locus, LOC102723944 rs6596830, rather than previously reported rs2745572, showed significant association with POAG (P = 8.61 × 10-4, odds ratio (OR) = 0.75) and HTG (P = 3.68 × 10-3, OR = 0.75). ABCA1 rs2487032 was also significantly associated with POAG (P = 3.00 × 10-5, OR = 0.70) and HTG (P = 2.08 × 10-4, OR = 0.70). Joint analysis showed that carriers of homozygous non-protective alleles of ABCA1 rs2487032 and LOC102723944 rs6596830 had 2.99-fold higher risk of POAG (P = 1.27 × 10-3) when compared to those carrying homozygous non-risk alleles. Patients with POAG carrying ABCA1 rs2487032 G allele had higher HDL cholesterol, and those with LOC102723944 rs6596830 A allele had lower LDL. This study revealed individual and joint association of ABCA1 and LOC102723944 variants with POAG in southern Chinese population. Subjects carrying non-protective alleles had increased risk to POAG, and corresponding genotypes would affect the lipid profiles.

Changes of Optic Disc and Macular Vessel Perfusion Density in Primary Angle Closure Glaucoma: A Quantitative Study Using Optical Coherence Tomography Angiograph.

INTRODUCTION: This study aims to investigate the changes of retinal vascular system in primary angle closure glaucoma (PACG) and acute primary angle closure (APAC) by optical coherence tomography (OCT) angiograph (OCTA) and to evaluate the diagnostic ability of changes of vessel density (VD) in different sectors and layers of optic disc and macular area in APAC and PACG. METHODS: In this cross-sectional, observational study, 21 APAC patients (22 eyes) and 21 PACG patients (27 eyes) along with 17 healthy people were enrolled from August 2018 to March 2019. Optic disc region and macular region were imaged using swept-source OCTA system. VD of the macular region was quantified by Image J (1.52a, USA) and Matlab 2018a. The circumpapillary retinal nerve fiber layer (cpRNFL) thickness and ganglion cell complex thickness were obtained by spectral-domain OCT. RESULTS: Compared with the healthy group, the cpRNFL thickness in superior sector was thicker in the APAC group, and this area had the most diffuse microvascular dropout as well. The difference in the macular superficial capillary plexus (SCP) VD between APAC and the control group was not statistically significant. The area under the ROC curves (AUC) of the total optic disc VD in the radial peripapillary capillary (RPC) layer was higher than the AUC of the papillary VD in the optic nerve head (ONH) layer. Compared to the control group, the total optic disc VD, peripapillary VD, and each quadrant of peripapillary VD were decreased in PACG (p < 0.01). In PACG macular region, SCP VD, and deep capillary plexus (DCP) VD, parafovea VD (except temporal sectors) decreased (p < 0.01). The PACG eyes had a greater decrease percentage of VD in total ONH than total macula. The diagnostic value of the VD in the ONH layer and the RPC layer was similar. The diagnostic value of the SCP VD in the macula was greater than the DCP VD in the macula. The AUC was no significant difference between cpRNFL thickness and the total optic disc VD AUC. CONCLUSION: Elevated intraocular pressure preferentially affects vascular perfusion in the optic disc region more than the macular region in APAC and PACG. In the APAC eyes, there was a perfusion defect in the optic disc region and an increase in RNFL thickness. In this study, the OCTA vascular parameters have similar performance to the OCT structural parameters for glaucoma diagnosis in PACG.

Sturge-Weber syndrome coexisting with polydactyly: a case report.

BACKGROUND: Sturge-Weber syndrome (SWS) is a sporadic congenital disorder, characterized by unilateral facial nevus flammeus associated with ipsilateral glaucoma, choroidal angioma and leptomeningeal hemangiomas. SWS can comorbid with other disorders in some patients, however, there has been no prior described case of SWS and polydactyly occurring in the same patient. CASE PRESENTATION: A 15-year-old girl with diagnosis of SWS presented to our hospital. She had bilateral glaucoma and extensive port-wine stains distributing in bilateral faces, left neck and left upper limb. Meanwhile, the patient was noted to demonstrate the superfluous digit attaching on the left thumb and was diagnosed as polydactyly. Trabeculectomy, with intraoperative application of mitomycin C and postoperative subconjunctival injections of 5-fluorouracil, was successful in controlling the intraocular pressure in both eyes. CONCLUSIONS: We report a case with bilateral SWS coexisting with unilateral polydactyly, which, to our knowledge, has not been recognized previously and adds further evidence to the existing literature. In view of the rare concurrence of SWS and polydactyly, the etiology is unclear and further investigation is required to explore the underlying pathogenesis.

Association of the CAV1-CAV2 locus with normal-tension glaucoma in Chinese and Japanese.

BACKGROUND: The CAV1-CAV2 locus has been associated with primary open-angle glaucoma (POAG) and intraocular pressure. However, its association with normal-tension glaucoma (NTG) was inconclusive. Therefore, we evaluated this association in Chinese and Japanese. METHODS: Two single-nucleotide polymorphisms (SNPs, rs4236601 and rs1052990) from previous genome-wide association studies of POAG were genotyped in a total of 2220 study subjects: a Hong Kong Chinese cohort of 537 NTG patients and 490 controls, a Shantou Chinese cohort of 102 NTG and 731 controls and an Osaka Japanese cohort of 153 NTG and 207 controls. Subgroup analysis by gender was conducted. Outcomes from different cohorts were combined using meta-analysis. RESULTS: SNP rs4236601 was significantly associated with NTG in the two Chinese cohorts (Pmeta = .0019, OR = 4.55, I2 = 0). In contrast, rs4236601 was monomorphic in the Osaka cohort. The association of rs1052990 was insignificant in a meta-analysis combining Chinese and Japanese cohorts (Pmeta = .81, OR = 1.05; I2 = 64%), and the OR tended towards opposite directions between Chinese (OR = 1.26) and Japanese (OR = 0.69). Gender-specific effects of the SNPs were not statistically significant in the logistic regression or Breslow-day tests of ORs (P > .05), although rs4236601 was significant in males (P = .0068; OR = 10.30) but not in females (P = .14; OR = 2.65) in the meta-analysis of Chinese subjects. CONCLUSIONS: In this study, we confirmed the association of rs4236601 at the CAV1-CAV2 locus with NTG in Chinese. SNP rs4236601 is monomorphic, and rs1052990 tends towards a different direction in the Japanese cohort. Further studies are warranted to verify the ethnic difference and gender-specific effects of this locus.

Caveolin-1 regulates human trabecular meshwork cell adhesion, endocytosis, and autophagy.

Impaired trabecular meshwork (TM) outflow is implicated in the pathogenesis of primary open-angle glaucoma (POAG). We previously identified the association of a caveolin-1 (CAV1) variant with POAG by genome-wide association study. Here we report a study of CAV1 knockout (KO) effect on human TM cell properties. We generated human CAV1-KO TM cells by CRISPR/Cas9 technology, and we found that the CAV1-KO TM cells less adhered to the surface coating than the wildtype TM cells by 69.34% ( P < 0.05), but showed no difference in apoptosis. Higher endocytosis ability of dextran and transferrin was also observed in the CAV1-KO TM cells (4.37 and 1.89-fold respectively, P < 0.001), compared to the wildtype TM cells. Moreover, the CAV1-KO TM cells had higher expression of extracellular matrix-degrading enzyme genes ( ADMTS13 and MMP14) as well as autophagy-related genes ( ATG7 and BECN1) and protein (LC3B-II) than the wildtype TM cells. In summary, results from this study showed that the CAV1-KO TM cells have reduced adhesion with higher extracellular matrix-degrading enzyme expression, but increased endocytosis and autophagy activities, indicating that CAV1 could be involved in the regulation of adhesion, endocytosis, and autophagy in human TM cells.

Association of the SIX6 locus with primary open angle glaucoma in southern Chinese and Japanese.

The purpose of the study was to evaluate the association profiles of the SIX6 locus with primary open-angle glaucoma (POAG) in southern Chinese and Japanese. In this study, we tested single marker and haplotype-based associations of 11 tagging single nucleotide polymorphisms (SNPs) covering the SIX6 locus with POAG in a Hong Kong Chinese cohort (N = 1402). A novel SNP (i.e., rs12436579) and two SNPs (i.e., rs33912345 and rs10483727) from previous genome-wide association studies were further tested in a Chinese cohort from Shantou (N = 888) and a Japanese cohort from Osaka (N = 463). Results from the three cohorts were meta-analysed using a random-effect model. We found rs12436579, which has not been previously reported, was associated with POAG in Hong Kong and Shantou Chinese (Pcombined = 4.3 × 10-5, OR = 0.72, I2 = 0). Additionally, we replicated the association of one known SNP, rs33912345 (Pcombined = 0.0061, OR = 0.69, I2 = 45%), with POAG in the Chinese cohorts but not in the Japanese cohort (P > 0.6). Another known SNP, rs10483727, was nominally associated with POAG in the two Chinese cohorts (Pcombined = 0.017, OR = 0.70, I2 = 53%). All these three SNPs were significantly associated with POAG when the three cohorts were combined in meta-analysis (Pcombined<0.005). Furthermore, two haplotypes, C-C (Pcombined = 1.13 × 10-5, OR = 1.41, I2 = 0) and A-A (Pcombined = 0.045, OR = 0.68, I2 = 70%), defined by rs33912345-rs12436579 were associated with POAG in Chinese but not in Japanese. In conclusion, this study confirmed the association between two GWAS SNPs in SIX6 (rs33912345 and rs10483727) and POAG. Also, a SNP, rs12436579, not associated with POAG before, was found to be associated with POAG in Chinese. Further studies are warranted to elucidate the role of this novel SNP in POAG.

Mutations of RagA GTPase in mTORC1 Pathway Are Associated with Autosomal Dominant Cataracts.

Cataracts are a significant public health problem with no proven methods for prevention. Discovery of novel disease mechanisms to delineate new therapeutic targets is of importance in cataract prevention and therapy. Herein, we report that mutations in the RagA GTPase (RRAGA), a key regulator of the mechanistic rapamycin complex 1 (mTORC1), are associated with autosomal dominant cataracts. We performed whole exome sequencing in a family with autosomal dominant juvenile-onset cataracts, and identified a novel p.Leu60Arg mutation in RRAGA that co-segregated with the disease, after filtering against the dbSNP database, and at least 123,000 control chromosomes from public and in-house exome databases. In a follow-up direct screening of RRAGA in another 22 families and 142 unrelated patients with congenital or juvenile-onset cataracts, RRAGA was found to be mutated in two unrelated patients (p.Leu60Arg and c.-16G>A respectively). Functional studies in human lens epithelial cells revealed that the RRAGA mutations exerted deleterious effects on mTORC1 signaling, including increased relocation of RRAGA to the lysosomes, up-regulated mTORC1 phosphorylation, down-regulated autophagy, altered cell growth or compromised promoter activity. These data indicate that the RRAGA mutations, associated with autosomal dominant cataracts, play a role in the disease by acting through disruption of mTORC1 signaling.

A common variant near TGFBR3 is associated with primary open angle glaucoma.

Primary open angle glaucoma (POAG), a major cause of blindness worldwide, is a complex disease with a significant genetic contribution. We performed Exome Array (Illumina) analysis on 3504 POAG cases and 9746 controls with replication of the most significant findings in 9173 POAG cases and 26 780 controls across 18 collections of Asian, African and European descent. Apart from confirming strong evidence of association at CDKN2B-AS1 (rs2157719 [G], odds ratio [OR] = 0.71, P = 2.81 × 10(-33)), we observed one SNP showing significant association to POAG (CDC7-TGFBR3 rs1192415, ORG-allele = 1.13, Pmeta = 1.60 × 10(-8)). This particular SNP has previously been shown to be strongly associated with optic disc area and vertical cup-to-disc ratio, which are regarded as glaucoma-related quantitative traits. Our study now extends this by directly implicating it in POAG disease pathogenesis.

The diagnostic ability of peripapillary vessel density in primary open-angle glaucoma suspects.

OBJECTIVE: To assess the diagnostic ability of peripapillary vessel density (pVD) in primary open-angle glaucoma suspect (GS) patients. METHODS: Sixteen primary open-angle GS patients (22 eyes) and 20 normal controls (22 eyes) were included. In the GS group, OCTA measurements of pVD (superior, inferior, nasal, temporal, and global), OCT measurements of retinal nerve fiber layer (RNFL) thickness, disc area, rim area and ganglion cell complex (GCC) thickness were examined. In the control group, pVD measurements were performed. The vessel density between the two groups was compared. The correlation between OCTA and OCT parameters was evaluated. The receiver operating characteristic curve (ROC) was used to evaluate the diagnostic efficacy of OCTA measurements. RESULTS: The global (P<0.001), nasal (P=0.003), and inferior (P=0.002) quadrant pVD in GS group was considerably lower than the control group. The global pVD was positively correlated with the inferior RNFL thickness (r=0.492, P=0.023) and rim area (r=0.483, P=0.027). The inferior pVD was positively correlated with the inferior RNFL thickness (r=0.648, P=0.001), the nasal RNFL thickness (r=0.441, P=0.045), the rim area (r=0.439, P=0.046) and the GCC thickness (r=0.472, P=0.048). The global pVD had the best diagnostic value (AUC=0.825, sensitivity 86.36%, specificity 72.73%, cutoff value 45%), followed by the inferior (AUC=0.749) and nasal (AUC=0.748) quadrant pVD. CONCLUSION: In primary open-angle GS patients, the global and inferior quadrant pVD was lower than that of normal people, and it was positively correlated with the inferior RNFL thickness and rim area. The diagnostic value of pVD for discriminating GS from normal people was excellent with high sensitivity and specificity.

Untargeted and Oxylipin-Targeted Metabolomics Study on the Plasma Samples of Primary Open-Angle Glaucoma Patients.

Purpose: to determine the metabolomics profiles in the plasma samples of primary open-angle glaucoma (POAG) patients. Methods: The plasma samples from 20 POAG patients under intraocular pressure (IOP)-lowering medication treatment and 20 control subjects were subjected to the untargeted metabolomics analysis, among which 10 POAG patients and 10 control subjects were further subjected to the oxylipin-targeted metabolomics analysis by liquid chromatography-mass spectrometry analysis. The prediction accuracy of the differentially abundant metabolites was assessed by the receiver operating characteristic curves. Pathway analysis and correlation analysis on the differentially abundant metabolites and clinical and biochemical parameters were also conducted. Results: Untargeted metabolomics profiling identified 33 differentially abundant metabolites in the POAG patients, in which the metabolism of linoleic acid, α-linolenic acid, phenylalanine, and tricarboxylic acid cycle were enriched. The correlation analysis indicated that the differentially abundant metabolites were associated with central corneal thickness, peripapillary retinal nerve fiber layer thickness, visual field defects, and lymphocytes. The oxylipin-targeted metabolomics analysis identified 15-keto-Prostaglandin F2 alpha, 13,14-Dihydro-15-keto-prostaglandin D2, 11-Dehydro-thromboxane B2, 8,9-Epoxyeicosatrienoic acid, and arachidonic acid to be significantly decreased in the POAG patients and enriched in the arachidonic acid (AA) pathway. Conclusions: This study revealed that the metabolites in the arachidonic acid metabolism pathway are differentially abundant, suggesting high IOP may not be the only detrimental factor for optic nerve cell damage in this group of POAG patients. Lipid metabolism instability-mediated alterations in oxylipins and AA pathways may be important in POAG, suggesting that oxidative stress and immune-related inflammation could be valuable directions for future therapeutic strategies.

Adeno-associated virus-mediated expression of growth-associated protein-43 aggravates retinal ganglion cell death in experimental chronic glaucomatous injury.

PURPOSE: To examine whether adeno-associated virus (AAV) vector-mediated overexpression of growth-associated protein-43 (GAP-43) has protective or deleterious effects on retinal ganglion cell (RGC) survival in laser-induced chronic intraocular pressure (IOP) elevation injury. METHODS: Adult Fischer 344 rats received unilateral intravitreal injection of either normal saline, AAV-green fluorescent protein (AAV-GFP), or a bicistronic AAV vector encoding GAP-43 and GFP (AAV-GAP-43). Two weeks later, experimental chronic glaucoma was induced in the injected eyes by scarring the trabecular meshwork with a diode laser. IOP was measured with an impact (rebound) tonometer. Survival of RGCs was estimated after 3 weeks of IOP elevation by quantifying ß-III tubulin⁺ neurons in retinal whole mounts. The transfection efficiency of target genes was assessed with direct view of GFP and western blot analysis of GAP-43. RESULTS: Quantification of ß-III tubulin⁺ immunostaining revealed that, compared to uninjured eyes (1,172±80 cells/mm²), 3 weeks of laser-induced IOP elevation led to a 60% decline in RGC survival (496±136 cells/mm²). Transfection with control vector AAV-GFP by itself did not have a significant effect on RGC viability (468±124 cells/mm²). Overexpression of GAP-43 in RGC cell bodies and axons via bicistronic AAV-GAP-43 led to more severe RGC death (260±112 cells/mm²) in IOP elevated eyes, an 80% loss of the total RGC population. CONCLUSIONS: Overexpression of GAP-43 aggravated RGC death in experimental chronic IOP elevation injury. GAP-43 was upregulated in RGCs regenerating after optic nerve injury. Thus, the finding that this same protein is deleterious to RGC viability after chronic IOP elevation may aid in understanding the mechanisms involved in RGC loss in glaucoma and how best to treat this condition.

The Diagnostic Value of Optical Coherence Tomography for Detecting Glaucoma in High Myopia.

PRCIS: Glaucomatous changes in high myopia are difficult to detect due to the similarity of functional and structural alteration in both disease. Optical coherence tomography (OCT) shows relatively high diagnostic accuracy in glaucoma with high myopia (HM). PURPOSE: This study aims to evaluate the thickness differences of OCT parameters between HM and HM with glaucoma (HMG), and to verify which parameters have a higher diagnostic value on this issue according to area under the receiver operating characteristic (AUROC) curve. MATERIALS AND METHODS: A comprehensive literature search was performed on PubMed, Embase, Medline, Cochrane, CNKI, and Wanfang. Eligible articles were identified by reviewing the retrieved results. The weighted mean difference and 95% CI for continuous outcomes and pooled AUROC were calculated. RESULTS: Fifteen studies with a total of 1304 eyes, including 569 high myopia and 735 HMG were comprised in this meta-analysis. Our results showed that, in comparison with HM, HMG had a significantly thinner thickness for retinal nerve fiber layer except for nasal sector, macular ganglion cell inner plexiform layer thickness except for superior sector and macular ganglion cell complex thickness. In contrast, the inferior sector and average thickness of retinal nerve fiber layer, macular ganglion cell complex and ganglion cell inner plexiform layer had relatively high AUROC. CONCLUSION: Based on the current study of retinal OCT measurement differences between HM and HMG, ophthalmologists should pay more attention to the thinning in inferior sector and the average thickness of macular and optic disc during the management of HM.

Relatively anterior lens position in primary angle-closure glaucoma eyes with long axial length.

Purpose: To evaluate the refractive status and ocular biometric parameters in primary angle-closure glaucoma (PACG) eyes with different axial lengths (ALs). Methods: In total, 742 Chinese PACG subjects with complete ophthalmic examinations were enrolled. The refractive status was categorized as myopia (spherical equivalent [SE] ≤-0.5 D), emmetropia (-0.5 D < SE < +0.5 D), and hyperopia (SE ≥+0.5 D), whereas the AL was divided into short (AL <22.5 mm), regular (22.5 ≤ AL <23.5 mm), and long (AL ≥23.5 mm). The refractive status and ocular biometric parameters were compared among different AL groups. Results: The mean AL of the PACG eyes was 22.53 ± 0.84 mm (range: 19.68-25.57 mm). The refractive status was significantly different among different AL groups (P < 0.001). Also, 92.6% of hyperopic PACG eyes showed AL <23.5 mm, and 19.0% of myopic PACG eyes showed AL ≥23.5 mm. The SE showed significant differences among different AL groups only in the hyperopic subjects (P = 0.012). The AL was significantly longer in myopic eyes (P < 0.001). The PACG eyes with longer AL exhibited lower keratometry, longer central anterior chamber depth and corneal diameter, and lens position and relative lens position closer to the anterior (P < 0.001). Conclusion: Axial hyperopia was common in PACG eyes, and axial myopia was not uncommon. Relatively anterior lens position could explain the occurrence of PACG in the eyes with long AL.

Association of body mass index and PXDNL gene variants with acute primary angle closure in southern Chinese population.

This study aimed to evaluate the association of body mass index (BMI) and the weight-related gene, peroxidasin-like (PXDNL), with acute primary angle closure (APAC) and primary angle-closure glaucoma (PACG) in southern Chinese population. Total 4700 study subjects (1024 APAC, 781 PACG, and 2895 control subjects) with complete ophthalmic examinations were enrolled into this study. The association of BMI with APAC, PACG and ocular biometric parameters was evaluated. Three PXDNL missense variants were genotyped by TaqMan assay, and their association with APAC and PACG was also investigated. Multivariable logistic regression analysis showed that BMI and body weight were significantly associated with both APAC and PACG (P < 0.01). Multiple linear regression analysis demonstrated that each 1 kg/m2 increased in BMI was associated with 0.038 mm increase in axial length, 0.018 mm increase in central anterior chamber depth, 0.002 mm increase in lens position, 0.012 mm increase in corneal diameter and 0.014 mm decrease in lens thickness among the APAC subjects (P < 0.001), but not with PACG. Genetic association analysis identified that PXDNL rs11985241-rs16916207 CT haplotype conferred a higher risk to APAC (OR = 1.25, P = 0.004) than the TG haplotype, but not with PACG. The APAC subjects carrying the rs11985241 C or rs16916207 T alleles showed significantly lower weight than those carrying the corresponding protective alleles. In summary, this study revealed that lower BMI could be associated with higher risk of APAC. PXDNL could be a new associated gene for APAC.

Evaluation of Myocilin Variant Protein Structures Modeled by AlphaFold2.

Deep neural network-based programs can be applied to protein structure modeling by inputting amino acid sequences. Here, we aimed to evaluate the AlphaFold2-modeled myocilin wild-type and variant protein structures and compare to the experimentally determined protein structures. Molecular dynamic and ligand binding properties of the experimentally determined and AlphaFold2-modeled protein structures were also analyzed. AlphaFold2-modeled myocilin variant protein structures showed high similarities in overall structure to the experimentally determined mutant protein structures, but the orientations and geometries of amino acid side chains were slightly different. The olfactomedin-like domain of the modeled missense variant protein structures showed fewer folding changes than the nonsense variant when compared to the predicted wild-type protein structure. Differences were also observed in molecular dynamics and ligand binding sites between the AlphaFold2-modeled and experimentally determined structures as well as between the wild-type and variant structures. In summary, the folding of the AlphaFold2-modeled MYOC variant protein structures could be similar to that determined by the experiments but with differences in amino acid side chain orientations and geometries. Careful comparisons with experimentally determined structures are needed before the applications of the in silico modeled variant protein structures.

A novel PAX6 deletion in a Chinese family with congenital aniridia.

PURPOSE: To identify a disease-causing paired box 6 (PAX6) gene mutation in a Chinese family affected by autosomal dominant congenital aniridia. METHODS: All participants in the study, including the aniridia family and 100 unrelated senile cataract controls, received a comprehensive ophthalmic examination. Genomic DNA was extracted from their whole blood. Mutation screen in all exons and their adjacent splicing junctions of PAX6 was performed by direct sequencing of polymerase chain reaction (PCR) products. PCR products of heterozygous mutation were further cloned into T-vectors and confirmed by sequencing. Multiple alignments were performed using ClustalX to compare PAX6 protein sequences among vertebrates. MicroRNA binding sites were predicted by TargetScan. RESULTS: A novel heterozygous PAX6 deletion c.1251_1353del103 (p.Pro418Serfs*87) affecting exon 14 and the 3'-untranslated-region (3'-UTR) was identified in the congenital aniridia family. The mutation was exclusively observed in all affected family members but not in any unaffected family member or unrelated control. Bioinformatics analysis showed that the deletion led to remarkable changes of the PAX6 protein, including a frameshift, changes of protein sequence, and a COOH-terminal extension. Multiple alignments showed that the affected region of PAX6 shared high sequence identity (100%) among its vertebrate orthologs. The COOH-terminal extension might also affect microRNA binding sites in the 3'-UTR as predicted by TargetScan. CONCLUSIONS: In the current study we reported a novel PAX6 deletion resulting in an abnormal PAX6 COOH-terminal extension in the Chinese family affected by aniridia. Our findings thus add to the mutation spectrum of PAX6.

Association analysis of cigarette smoking with onset of primary open-angle glaucoma and glaucoma-related biometric parameters.

BACKGROUND: To date, studies on the role played by cigarette smoking in primary open-angle glaucoma (POAG) remains controversial. The current study evaluated cigarette smoking as a risk factor of POAG and its relationships with vertical cup-to-disc ratio (VCDR), central corneal thickness (CCT) and intraocular pressure (IOP) in a Chinese cohort. METHODS: In a total of 248 unrelated individuals including 30 juvenile-onset POAG (JOAG), 92 adult-onset POAG (AOAG) and 126 sex-matched senile cataract controls, underwent comprehensive ophthalmic examination. Their smoking was obtained and documented by questionnaire. Association of cigarette smoking with POAG was performed using logistic regression controlled for age and sex. Effects of cigarette smoking on VCDR, IOP and CCT were analyzed with multiple linear regression. RESULTS: In either JOAG or AOAG, no association of cigarette smoking was found with disease onset (P = 0.692 and 0.925 respectively). In controls and JOAG, no significant effects of smoking were found on VCDR, IOP or CCT (all P > 0.05). Smoking was found to be correlated with decreased CCT in AOAG and combined POAG (JOAG + AOAG) (P = 0.009 and 0.003), but no association with VCDR or IOP was observed (P > 0.05). CONCLUSIONS: Although cigarette smoking was not found to be risk factor for onset of POAG, it was correlated with CCT in AOAG, and thus might still play a role in the disease course, especially for AOAG.

Determinants of peripapillary retinal nerve fiber layer's grayscale value in normal eyes by spectral domain optical coherence tomography.

To determine and evaluate the distribution, variation, and determinants of peripapillary retinal nerve fiber layer (pRNFL) grayscale value with spectral-domain optical coherence tomography (SD-OCT) in normal eyes. In this cross-sectional study, three hundred ninety-seven normal eyes from 397 healthy Chinese adults aged 18-80 were consecutively recruited from a tertiary eye care center. An SD-OCT instrument took pRNFL imaging. We used a customized software to measure pRNFL parameters, including thickness and grayscale value. Univariable and multiple linear regression analyses were performed to examine the relationship between pRNFL grayscale value with ocular (e.g., axial length [A.L.], spherical equivalent [S.E.], intraocular pressure [IOP]), and systemic (e.g., age, sex) factors. A total of 397 eyes from 397 healthy subjects were included in the final analysis with mean (± SD) age 44.63 ± 16.43 years (range 18-80 years) and 196 (49.4%) males. The mean average of pRNFL grayscale value and thickness 164.82 ± 5.69 and 106.68 ± 8.89 µm, respectively. pRNFL grayscale value in nasal sectors (163.26 ± 9.31) was significantly lower comparing those in all other five sectors (all with p < 0.001)]. In multivariable analysis, average pRNFL grayscale value was independently correlated to older age (ß = - 0.053, p = 0.002), longer axial length (ß = - 0.664, p = 0.003), lower RPE grayscale value (ß = 0.372, p < 0.001) and lower ImageQ (ß = 0.658, p < 0.001). In this study, we provided normative SD-OCT data on the pRNFL grayscale value profile in nonglaucomatous eyes. Lower average pRNFL grayscale value was independently correlated to older age, longer axial length, lower RPE grayscale value, and lower ImageQ. These determinants should be considered when interpreting pRNFL grayscale value in glaucoma assessment.

Development and Evaluation of the Prognostic Nomogram to Predict Refractive Error in Patients With Primary Angle-Closure Glaucoma Who Underwent Cataract Surgery Combined With Goniosynechialysis.

Objective: To evaluate the accuracy of different intraocular lens (IOL) power calculation formulas and develop prognostic nomograms to predict the risk of postoperative refractive error in primary angle-closure glaucoma (PACG) patients. Methods: A total of 111 eyes with PACG underwent goniosynechialysis combined with phacoemulsification and IOL implantation were included. SRK/T, Barrett II, Hoffer Q, and Kane formulas were used to predict postoperative refraction. Prediction error (PE) and absolute predictive error (APE) produced by the four formulas were calculated and compared. An APE >0.50 D was defined as the event. Binary logistic regression analysis and prognostic nomogram models were conducted to investigate reliable predictors associated with postoperative refraction. Results: The Kane (-0.06 D) and Barrett II (-0.07 D) formulas had mean prediction error close to zero (p = 0.44, p = 0.41, respectively). The Hoffer Q and SRK/T produced significantly myopic outcomes (p = 0.003, p = 0.013, respectively). The percentage of eyes within ± 0.5 D was 49.5% (55/111), 44.1% (49/111), 43.2% (48/111), and 49.5% (54/111), for the Kane, Barrett II, Hoffer Q, and SRK/T formula, respectively. Nomogram showed that AL had the greatest impact on the refractive outcomes, indicating a shorter preoperative AL is associated with a greater probability of refractive error event. The area under the receiver operator curve (AUC) of the nomogram for the Kane, Barrett II, Hoffer Q, and SRK/T was 0.690, 0.701, 0.708, and 0.676, respectively. Conclusions: The Kane and Barrett II formulas were comparable, and they outperformed Hoffer Q and SRK/T in the total eyes with PACG receiving cataract surgery combined with goniosynechialysis. The developed nomogram models can effectively predict the occurrence of postoperative refractive error events.