Comparative Proteomic Analysis of Aqueous Humor Reveals Biochemical Disparities in the Eyes of High Myopic Patients.

Myopia accounts for a significant proportion of visual lesions worldwide and has the potential to progress toward pathological myopia. This study aims to reveal the difference in protein content in aqueous humor between high myopic and nonhigh myopic patients, as well as better understand the dysregulation of proteins in myopic eyes. Aqueous humor was collected for liquid chromatograph mass spectrometer (LC/MS) analysis from 30 individual eyes that underwent phacoemulsification and intraocular lens (IOL) implantation. Results showed that a total of 190 differentially expressed proteins were identified, which revealed their involvement in cell metabolism, immune and inflammatory response, and system and anatomical structure. Further analysis focused on 15 intensively interacted hub proteins, encompassing functions related to complement cascades, lipoprotein metabolism, and fibrin biological function. Subsequent validations demonstrated elevated levels of APOE (apolipoprotein E), C3 (complement 3), and AHSG (α-2-HS-glycoprotein) in the high myopia group (31 eyes of cataracts and 45 eyes of high myopia with cataracts). AHSG had a significant positive correlation with axial length in high myopic patients, with good efficacy in distinguishing between myopic and nonmyopic groups. AHSG may be a potential indicator of the pathological severity and participator in the pathological progress of high myopia. This study depicted differential expression characteristics of aqueous humor in patients with high myopia and provided optional information for further experimental research on exploring the molecular mechanisms and potential therapeutic targets for high myopia. Data are available via ProteomeXchange with the identifier PXD047584.

Wnt5a/ß-catenin-mediated epithelial-mesenchymal transition: a key driver of subretinal fibrosis in neovascular age-related macular degeneration.

BACKGROUND: Neovascular age-related macular degeneration (nAMD), accounts for up to 90% of AMD-associated vision loss, ultimately resulting in the formation of fibrotic scar in the macular region. The pathogenesis of subretinal fibrosis in nAMD involves the process of epithelial-mesenchymal transition (EMT) occurring in retinal pigment epithelium (RPE). Here, we aim to investigate the underlying mechanisms involved in the Wnt signaling during the EMT of RPE cells and in the pathological process of subretinal fibrosis secondary to nAMD. METHODS: In vivo, the induction of subretinal fibrosis was performed in male C57BL/6J mice through laser photocoagulation. Either FH535 (a ß-catenin inhibitor) or Box5 (a Wnt5a inhibitor) was intravitreally administered on the same day or 14 days following laser induction. The RPE-Bruch's membrane-choriocapillaris complex (RBCC) tissues were collected and subjected to Western blot analysis and immunofluorescence to examine fibrovascular and Wnt-related markers. In vitro, transforming growth factor beta 1 (TGFß1)-treated ARPE-19 cells were co-incubated with or without FH535, Foxy-5 (a Wnt5a-mimicking peptide), Box5, or Wnt5a shRNA, respectively. The changes in EMT- and Wnt-related signaling molecules, as well as cell functions were assessed using qRT-PCR, nuclear-cytoplasmic fractionation assay, Western blot, immunofluorescence, scratch assay or transwell migration assay. The cell viability of ARPE-19 cells was determined using Cell Counting Kit (CCK)-8. RESULTS: The in vivo analysis demonstrated Wnt5a/ROR1, but not Wnt3a, was upregulated in the RBCCs of the laser-induced CNV mice compared to the normal control group. Intravitreal injection of FH535 effectively reduced Wnt5a protein expression. Both FH535 and Box5 effectively attenuated subretinal fibrosis and EMT, as well as the activation of ß-catenin in laser-induced CNV mice, as evidenced by the significant reduction in areas positive for fibronectin, alpha-smooth muscle actin (α-SMA), collagen I, and active ß-catenin labeling. In vitro, Wnt5a/ROR1, active ß-catenin, and some other Wnt signaling molecules were upregulated in the TGFß1-induced EMT cell model using ARPE-19 cells. Co-treatment with FH535, Box5, or Wnt5a shRNA markedly suppressed the activation of Wnt5a, nuclear translocation of active ß-catenin, as well as the EMT in TGFß1-treated ARPE-19 cells. Conversely, treatment with Foxy-5 independently resulted in the activation of abovementioned molecules and subsequent induction of EMT in ARPE-19 cells. CONCLUSIONS: Our study reveals a reciprocal activation between Wnt5a and ß-catenin to mediate EMT as a pivotal driver of subretinal fibrosis in nAMD. This positive feedback loop provides valuable insights into potential therapeutic strategies to treat subretinal fibrosis in nAMD patients.

APOC1 is a prognostic biomarker associated with M2 macrophages in ovarian cancer.

BACKGROUND: Recent studies have demonstrated that APOC1 is associated with cancer progression, exerting cancer-promoting and immune infiltration-promoting effects. Nevertheless, there is currently no report on the presence of APOC1 in ovarian cancer (OV). METHOD: In this study, we conducted data analysis using the GEO and TCGA databases. We conducted a thorough bioinformatics analysis to investigate the function of APOC1 in OV, utilizing various platforms including cBioPortal, STRING, GeneMANIA, LinkedOmics, GSCALite, TIMER, and CellMarker. Additionally, we performed immunohistochemical staining on tissue microarrays and conducted in vitro cellular assays to validate our findings. RESULT: Our findings reveal that APOC1 expression is significantly upregulated in OV compared to normal tissues. Importantly, patients with high APOC1 levels show a significantly poorer prognosis. Furthermore, our study demonstrated that APOC1 exerted a crucial function in promoting the capacity of ovarian cancer cells to proliferate, migrate, and invade. Additionally, we have identified that genes co-expressed with APOC1 are primarily associated with adaptive immune responses. Notably, the levels of APOC1 in OV exhibit a correlation with the presence of M2 Tumor-associated Macrophages (TAMs). CONCLUSION: APOC1 emerges as a promising prognostic biomarker for OV and exhibits a significant association with M2 TAMs in OV.

SPARC-YAP/TAZ inhibition prevents the fibroblasts-myofibroblasts transformation.

BACKGROUND: Fibrotic scar is a severe side effect of trabeculectomy, resulting in unsatisfactory outcomes for glaucoma surgery. Accumulating evidence showed human Tenon's fibroblasts (HTFs) play an important role in fibrosis formation. We previously reported that the aqueous level of secreted protein acidic and rich in cysteine (SPARC) was higher in the patients with primary angle closure glaucoma, which was associated with the failure of trabeculectomy. In this study, the potential effect and mechanism of SPARC in promoting fibrosis were explored by using HTFs. METHODS: HTFs were employed in this study and examined under a phase-contrast microscope. Cell viability was determined by CCK-8. The expressions of SPARC-YAP/TAZ signaling and the fibrosis-related markers were examined with reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), Western blot, and immunofluorescence, subcellular fractionation was conducted to further determined the variation of YAP and phosphorylated YAP. The differential gene expressions were analyzed with RNA sequencing (RNAseq), followed by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. RESULTS: Exogenous SPARC induced HTFs-myofibroblast transformation, as evidenced by the increased expression of α-SMA, collagen I and fibronectin in both protein and mRNA levels. SPARC knockdown decreased the expressions of the above genes in TGF-ß2-treated HTFs. KEGG analysis showed that the Hippo signaling pathway was mostly enriched. SPARC treatment increased the expressions of YAP, TAZ, CTGF and CYR61 as well as enhanced YAP translocation from cytoplasm to nucleus, and decreased the phosphorylation of YAP and LAST1/2, which was reversed by SPARC knockdown. Knockdown of YAP1 decreased the fibrosis-related markers, such as α-SMA, collagen I and Fibronectin, in SPARC-treated HTFs. CONCLUSIONS: SPARC induced HTFs-myofibroblast transformation via activating YAP/TAZ signaling. Targeting SPARC-YAP/TAZ axis in HTFs might provide a novel strategy for inhibiting fibrosis formation after trabeculectomy.

Vault-Correlated Efficacy and Safety of Implantable Collamer Lens V4c Implantation for Myopia in Patients with Shallow Anterior Chamber Depth.

INTRODUCTION: The aim of the study was to evaluate efficacy and safety outcomes after implantation of the Visian Implantable Collamer Lens (ICL V4c) in myopia patients with shallow anterior chamber depth (ACD). METHODS: This retrospective study followed 163 eyes of 94 patients for at least 24 months. Uncorrected distance visual acuity (UDVA), corrected distance visual acuity, intraocular pressure (IOP), manifest refraction, vault, endothelial cell density (ECD), anterior chamber angle (ACA), anterior chamber volume, and the distance from the corneal endothelium to the central ICL (C-ICL) were measured during follow-ups. Spearman's correlation and logistic regression were used to identify variables correlated with changes in ECD and potential risk factors for ineffective outcomes, respectively. RESULTS: All surgeries were performed safely. High IOP of 9 eyes and anterior capsular opacity of 5 eyes were observed. The last follow-up ACA had a significant difference between the high and normal IOP groups (p = 0.0003). The mean ECD and vault were 2,855.76 ± 270.82 cells/mm2 and 388.01 ± 135.28 µm at the last follow-up, respectively. The vault and C-ICL were significantly associated with ΔECD (all p < 0.05). Furthermore, the vault was most responsible for the ECD loss. Twenty-two eyes had unsatisfactory postoperative UDVA, and the low vault at the last follow-up was a significant risk factor for this ineffective outcome (p < 0.001, OR = 14.739). CONCLUSIONS: ICL V4c implantation in patients with shallow ACD achieved stable visual outcomes. The vault is related to postoperative visual acuity and ECD loss, which needs to be paid attention during follow-up.

Efficacy and Safety of Implantable Collamer Lens V4c Implantation in 1,834 Myopic Eyes for 1 Year of Follow-up.

PURPOSE: To evaluate visual outcomes of patients with myopia after EVO Implantable Collamer Lens (ICL) (STAAR Surgical) implantation and predict risk factors of postoperative vault abnormalities. METHODS: In this single-center, retrospective analysis, 1,834 eyes of 926 patients with myopia who underwent EVO ICL implantation were recruited between 2020 and 2021. Patients were followed up for 1 year, during which surgery outcomes were evaluated. In addition, 31 eyes with vault abnormalities who underwent secondary surgery were enrolled to form a generalized linear model, which aimed to predict risk factors contributing to vault abnormalities. RESULTS: At the final follow-up visit, safety and efficacy indexes were 1.12 ± 0.17 and 1.10 ± 0.19, respectively, and there was no statistical significance between the low and high myopia groups. Furthermore, 79.18% of eyes achieved a residual spherical equivalent within ±0.50 diopters. Except for the temporary elevation of intraocular pressure at 1 week postoperatively, IOP and endothelial cell density remained stable during follow-up visits. The rate of postoperative adverse events was 21.97% and most adverse events were transient. Vault abnormalities accounted for the majority of complications (9.54%). Results of generalized linear model showed that age was a risk factor for postoperative vault abnormalities, and the anterior chamber depth served as a protective factor (P < .05). CONCLUSIONS: Visual outcomes of EVO ICL implantation were satisfactory in safety and efficacy indexes in both the low and high myopia groups for 1 year of follow-up, with acceptable stability in postoperative spherical equivalent, intraocular pressure, and endothelial cell density. This study emphasized cautious ICL size selection for older patients and those with shallow anterior chamber depth. [J Refract Surg. 2023;39(10):694-704.].

Aqueous Humor Growth Factor Levels and Trabeculectomy Outcomes in Primary Open-Angle Glaucoma Patients: A 2-Year Prospective Study.

Purpose: Maintenance of a filtering bleb is essential for long-term intraocular pressure control after trabeculectomy. Surgical site fibrosis and excessive extracellular matrix production are common causes of trabeculectomy failure, mediated by several growth factors. We aimed to evaluate the levels of five growth factors and their correlation with trabeculectomy outcomes in patients with primary open-angle glaucoma (POAG). Methods: We collected aqueous humor samples intraoperatively from patients with POAG who underwent trabeculectomy and measured the concentrations of transforming growth factor-ß (TGF-ß), acidic fibroblast growth factor (aFGF), insulin-like growth factor-1, vascular endothelial growth factor, and platelet-derived growth factor using multiplexed immunoassay kits. Intraocular pressure was measured with Goldmann applanation tonometry at 1 week and at 1, 3, 6, 12, 18, and 24 months after trabeculectomy. We allocated the eyes based on surgical outcome into a success or failure group. Results: Significantly high levels of aFGF and TGF-ß were observed in the failure group (both P < 0.0001) and were significant risk factors for trabeculectomy outcomes. Higher success rates were observed over the 24-month follow-up period in eyes with low aFGF and TGF-ß levels compared to eyes with high levels (P = 0.0031 and P = 0.0007, respectively). The levels of TGF-ß were significantly positively correlated with aFGF. Conclusions: In POAG patients, high aFGF and TGF-ß levels were significant risk factors for trabeculectomy failure. Translational Relevance: Modulation of aFGF and TGF-ß expression may have potential clinical applications after filtration surgery.