Molecular Vision最新文献

Purpose: The purpose of this cross-sectional study was to investigate the relationship between oxidation balance scores (OBSs) and myopia.

Methods: Participant information came from the National Health and Nutrition Examination Survey (NHANES) 2007-2008. The relationship between OBSs and myopia was analyzed using a restricted cubic spline, generalized linear regression, trend analysis, and ordinal logistic regression. The important components of the OBS in myopia were analyzed using XGBoost, random forest, and AdaBoost.

Results: The data of 5,187 participants from NHANES were collected, and a preliminary analysis was conducted. We found that an association between OBSs and myopia was only found in participants aged ≥ 20 years (n = 4,253). There was a linear relationship between OBSs and the occurrence of myopia in them. The logistic regression showed that OBSs were correlated with an increased incidence of myopia after adjusting for all confounders (OR: 1.01, 95% CI [1.00, 1.02]). The trend test showed that the higher the OBS, the higher the likelihood of developing myopia (p for trend < 0.05). There was a nonlinear relationship between OBSs and myopia severity according to a generalized additive model (β = 0.01, 95% CI [0.00, 0.01], p < .01). The ordered logistic regression analysis showed that for every unit increase in OBS, the likelihood of myopia severity increased by 11% after adjusting for all confounders. We also found that calcium was an important OBS component related to the incidence of myopia.

Conclusions: OBS is positively associated with the occurrence and severity of myopia in adults ≥ 20 years of age, and calcium is an important OBS component related to the incidence of myopia.

Purpose: This study aimed to determine the role of peroxisome proliferator-activated receptor α (PPARα) on corneal epithelial wound healing.

Methods: Ten-week-old PPARα knockout (PPARα^-/- ) mice and wild-type (WT) C57BL/6 mice and ex vivo cultured human corneal epithelial cells were used to investigate the function of PPARα on corneal epithelial wound healing. A two-millimeter diameter of the mice's central corneal epithelium was removed to induce corneal epithelial injury. The expression of PPARα during corneal epithelial wound healing was analyzed using immunofluorescent staining and quantitative RT-PCR. Histological and immunostaining techniques were used to evaluate corneal morphology, cell proliferation, and inflammatory response in WT and PPARα^-/- mice. PPARα agonist fenofibrate was used to determine its effect on corneal epithelial wound healing.

Results: PPARα expression was found to significantly increase during corneal epithelial repair. PPARα^-/- mice exhibited delayed corneal epithelial wound healing compared to WT mice. PPARα^-/- mice displayed altered proliferative responses and distinct patterns of inflammatory infiltrates. Administration of fenofibrate to WT mice resulted in accelerated corneal epithelial repair and increased PPARα expression and cell proliferation. In vitro studies using human corneal epithelial cells further supported the impact of fenofibrate on promoting corneal epithelial cell wound healing.

Conclusions: PPARα is a regulator of corneal epithelial wound healing, and its absence leads to delayed repair processes in the corneal epithelium.

Purpose: North Carolina macular dystrophy (NCMD) is a rare autosomal dominantly inherited congenital maculopathy caused by either non-coding point mutations or tandem duplications in the DNase I hypersensitivity site DHS6S1, at chromosome 6q16 (MCDR1), or at chromosome 5 (MCDR3). To date, at least 30 NCMD pedigrees from different ethnicities have been genetically identified worldwide. Herein, we report the clinical and genetic features of a newly found NCMD family in Mexico with a novel tandem duplication involving both the DNASE1 site and the PRDM13 gene.

Methods: Seven affected subjects from a Mexican family underwent a complete ophthalmic assessment that included dilated indirect ophthalmoscopy, fundus photography, optical coherence tomography (OCT), fundus autofluorescence (FAF), kinetic and chromatic perimetry, and electroretinography (ERG). Next-generation sequencing (NGS), followed by array-based comparative genomic hybridization (array-CGH) and quantitative polymerase chain reaction (qPCR) analyzes, were employed to demonstrate the causative molecular defect.

Results: All seven affected patients had a severe appearing phenotype characterized by symmetric excavated atrophic coloboma-like chorioretinal macular lesions. In addition, using OCT, lacunae in the inner retinal layers and inner retinal loss were observed in all patients. NGS identified a heterozygous tandem duplication of the entire coding sequence of the PRDM13 gene in all seven affected individuals, whereas subsequent array CGH, NGS, and Sanger sequencing allowed for the identification of the precise boundaries of a ~148 kb MCDR1 duplication containing the whole PRMD13 gene and the DNASE1 site.

Conclusions: The phenotypic features in this NCMD pedigree continue to support the concept that this disorder is a congenital macular malformation rather than a progressive dystrophic entity. Unlike most NCMD families, there was no variable expressivity found in this study, possibly due to the relatively small size of the family. The other hypothesis is that the duplication involves genomic segments that are more consistently or tightly bound to other regulatory regions of PRDM13. The identification of a novel causative tandem duplication involving the DNASE1 site and the PRDM13 gene in this family allows for the expansion of the mutational spectrum of the disease.

Purpose: Diabetic patients experience chronic hyperglycemia that increases oxidative stress by enhancing free radical formation and nitric oxide (NO) production. Genetic mutations in the endothelial nitric oxide synthase (eNOS) enzyme gene affect the levels of NO formation. These mutations, together with chronic hyperglycemia, may increase the risk of diabetic retinopathy (DR) development and/or DR progression as a complication of diabetes. This study aimed to determine whether the eNOS polymorphisms intron 4ab, exon 7 Glu298Asp variant (G894T), and T-786C are associated with DR severity.

Methods: This case-control study involved 250 subjects (172 with type 2 diabetes mellitus (DM) with or without DR and 78 healthy controls). DR was detected by slit lamp biomicroscopy and its severity was determined with the evidence-based International Clinical Diabetic Retinopathy Disease Severity Scale. The genotyping of eNOS polymorphisms was analyzed by polymerase chain reaction (PCR) only or with restriction fragment length polymorphism. The haplotype analysis was performed using the SNPStats tool.

Results: The genotype distribution for the three polymorphisms was significantly different in patients with diabetes compared to controls (intron 4 ab: a/a, 1.7%; a/b, 20.4%; b/b, 77.9%. G894T: GG, 56.4%; GT, 34.3%; TT, 9.3%. T-786C: TT, 58.2%; TC, 33.5%; CC, 8.3%). Differences in genotype or allele frequency was non-significant between subjects with diabetes and DR compared to those without DR, except the C allele of the T-786C polymorphism was significantly less common in DR patients. DR severity was not affected by any polymorphisms. Haplotypes bTC and aTT were significantly less common or more prevalent in DR than DM patients, respectively.

Conclusions: We demonstrated that type 2 DM patients exhibited a higher prevalence of the three polymorphisms when compared to the control group. The C allele of T-786C polymorphism may have a protective effect against DR. Also, none of the mutations was correlated with a higher DR risk nor with the severity of DR. Haplotype aTT increased the risk for DR, while the bTC haplotype reduced it.

The circadian clock, a conserved biologic timekeeping mechanism, is pivotal in orchestrating rhythmic physiologic processes. While extensively studied in the central clock, the involvement of BMAL1 in peripheral clocks, particularly in human Müller cells, remains underexplored. Müller cells, critical for retinal homeostasis, may unveil novel insights into circadian regulation. Employing ChIP-sequencing, we comprehensively mapped BMAL1 binding sites in human Müller cells. The analysis identified 275 reproducible peaks, with predominant distribution across promoters (26.6%), intronic (26.3%), and intergenic (22.1%) regions, with 80% of these confident peaks linked to protein-coding genes. Differential peak analysis revealed 89 unique genes significantly enriched with BMAL1 sites in their promoters, while functional enrichment of the associated genes indicated key biologic processes such as circadian regulation of gene expression, photoperiodism, and glucocorticoid receptor signaling pathway regulation. Motif analysis revealed a highly conserved 6-nucleotide motif, CACGTG, appearing in 89.09% of the peaks. Analysis of the binding sites across genomic regions highlighted the robust BMAL1 binding, further confirmed by qPCR validation of circadian targets such as G6PC3, CIART, PER1, and TXNIP, which are critical for Müller cell health, along with SHMT2 and MALAT1, which have emerged as novel genes that may have implications for Müller cell health. Our findings unveil the regulatory landscape of BMAL1 in Müller cells, contributing to a broader understanding of the clock-mediated mechanism in ocular tissues. These insights hold therapeutic potential for circadian-related retinal diseases, presenting avenues for chronotherapeutic interventions.

Purpose: To explore the role of cytokines during the progression process of cataract patients with pathologic myopia (PMC) and simple high myopia (SHMC).

Methods: A total of 63 cataract patients who underwent cataract surgery were classified into a PMC group (22 eyes), an SHMC group (21 eyes), and an age-related cataract (ARC) group (20 eyes), based on axial length (AL) and International Myopia Institute (IMI)'s classification. Aqueous humor samples were extracted before surgery. Cytometric bead array (CBA) was employed to measure the level of interleukin-6 (IL-6), vascular endothelial growth factor (VEGF), interleukin-8 (IL-8), transforming growth factor-β1 (TGF-β1), basic fibroblast growth factor (bFGF), interleukin-10 (IL-10), interleukin-17a (IL-17a), interleukin-1β (IL-1β), monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor -α (TNF-α), intercellular adhesion molecule (ICAM), and vascular cell adhesion molecule (VCAM). Additionally, the correlations between cytokines and the AL or myopic maculopathy categories were examined.

Results: VEGF, IL-6, MCP-1, ICAM, and VCAM (all p<0.001), TGF-β1 (p=0.018), and IL-8 (p=0.008) were statistically different among the three groups. In parallel, the levels of VCAM (r=0.718), MCP-1 (r=0.591), ICAM (r=0.584), IL-8 (r=0.435), IL-6 (r=0.396), and TNF-α (r=0.280) were positively associated with myopic maculopathy, while VEGF (r=-0.542), TGF-β1 (r=-0.381), and IL-17a (r=-0.284) were correlated inversely with myopic maculopathy (all p<0.05). Furthermore, a significant positive correlation was observed between AL and levels of VCAM (r=0.726), MCP-1 (r=0.644), ICAM (r=0.573), IL-6 (r=0.386), and IL-8(r=0.376). VEGF (r=-0.610), TGF-β1 (r=-0.361), and IL-17a (r=-0.319) were inversely associated with AL (all p<0.05). Further analysis using multiple regression indicated that, after adjusting for confounding factors, lower VEGF and higher VCAM were significantly associated with AL. However, the limitations of this study were reflected in the inability to determine whether the changes in cytokines were the consequences or causes of the formation of high myopia.

Conclusions: The pathogeneses of PMC and SHMC may differ, and there are significant changes associated with inflammation and the immune response in eyes with PMC.

Purpose: Neovascular age-related macular degeneration (nAMD) is now a major cause of central vision loss in older adults worldwide. The primary characteristic of nAMD is the formation of macular neovascularization (MNV), which is a pathologic form of angiogenesis. Epigenetics plays a role in multiple pathological physiologic processes. N6-methyladenosine (m6A) modification is the most common, abundant, and reversible modification in eukaryotic mRNAs, and it plays a role in various pathological angiogenesis processes. This study intends to reveal the expression and functions of m6A during the macular neovascularization (MNV) process.

Methods: A laser-induced MNV mouse model was used in this study. m6A quantitative analysis was performed to detect the expression of m6A. Subsequently, the expression of various m6A writers and erasers was detected using quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. Immunohistochemistry was used to detect Wilms' tumor 1-associating protein (WTAP) expression in the MNV lesions. Intravitreal injection of WTAP siRNA in MNV mice to silence the WTAP gene. Hematoxylin and eosin (H&E) were used to determine the thickness and length of the MNV. Fundus fluorescein angiography (FFA) and indocyanine green angiography (ICGA) were examined to measure the leakage area of the MNV. Proliferating cell nuclear antigen (PCNA) expression was detected with a western blot. The mRNA and protein levels of β-catenin were tested with qRT-PCR and western blot.

Results: We found increased m6A modification levels after laser induction compared with the normal control group. Subsequently, the expression of various m6A writers and erasers was detected. The results showed that WTAP increased in the MNV model in mice. After the injection of WTAP siRNA into the vitreous body, the expression of WTAP significantly decreased, subsequently decreasing the m6A modification levels. The width, breadth, and leakage area of MNV damage markedly decreased, and endothelial cell proliferation was inhibited. After laser-induced MNV, the expression of β-catenin increased, and that of β-catenin significantly decreased after WTAP knockout.

Conclusions: In conclusion, this study suggests that WTAP-mediated m6A methylation can regulate pathological angiogenesis during MNV and that WTAP may participate in the formation of MNV through the wingless-related integration site (Wnt) pathway. WTAP may be a potential target for MNV treatment.

Purpose: Cataracts are typically treated by phacoemulsification followed by intraocular lens implantation. Studies of mouse models of cataract surgery have revealed that lens epithelial cells rapidly remodel their transcriptome to express proinflammatory cytokines after lens fiber cell removal, but it is currently unknown whether this response is conserved in human lenses. This study seeks to fill this knowledge gap.

Methods: Human cadaver eyes from 70 to 89 year old individuals were prepared for the human capsular bag model of cataract surgery. The central epithelium was preserved in RNAlater during culture preparation, then the equatorial epithelium was either immediately preserved in RNAlater after the culture was created, or 24 h later. Gene expression profiles were generated by bulk sequencing of RNA isolated from these tissue samples. The transcriptomic response of human cadaver-derived lens epithelial cells to culture in this "capsular bag" model was characterized by bioinformatic analysis. The human response was directly compared to that of 24-month-old mouse lens epithelial cells subjected to fiber cell removal surgery.

Results: Human lens epithelial cells remodel approximately a third of their transcriptome by 24 h after surgery, and like mice, this response consists of induction of proinflammatory cytokine genes, upregulation of fibrotic markers and downregulation of genes controlling the lens epithelial phenotype.

Conclusions: These observations demonstrate that humans and mice have similar responses to cataract surgery and support the use of mice to study the response of lens epithelial cells to cataract surgery, suggesting that identified injury response mechanisms can be leveraged to elucidate new approaches to improve the outcomes of cataract surgery.