Endothelial Tyrosine Kinase Tie1 Is Required for Normal Schlemm’s Canal Development

Background: Schlemm’s canal (SC) is a large vessel residing in the iridocorneal angle and is required to regulate aqueous humor outflow. Normal SC structure and function is indispensable for maintaining normal intraocular pressure, and elevated intraocular pressure is a risk factor for development of glaucoma. Recent reports have identified a key role of the angiopoietin-Tie2 pathway for SC development and function; however, the role of the orphan receptor Tie1 has not been clarified. Methods: We used Tie1 knock out mice to study the function of Tie1 in SC development and function. Real-time quantitative polymerase chain reaction and Western blot analyses were used to verify Tie1 deletion. High-resolution microscopy of mouse SC whole mount and cross sections were used to study SC morphology. Measurement of intraocular pressure in live mice was used to study the impact of Tie1 on SC function. Results: Tie1 is highly expressed in both human and mouse SC. Tie1 knock out mice display hypomorphic SC and elevated intraocular pressure as a result of attenuated SC development. Conclusions: Tie1 is indispensable for SC development and function, supporting it as a novel target for future SC-targeted glaucoma therapies and a candidate gene for glaucoma in humans.

360° Ab-Interno Schlemm’s Canal Viscodilation with OMNI Viscosurgical Systems for Open-Angle Glaucoma—Midterm Results

Purpose: To evaluate the effectiveness of ab-interno microcatheterization and 360° viscodilation of Schlemm’s canal (SC) performed with OMNI viscosurgical system in open angle glaucoma (OAG) together or not with phacoemulsification. Setting: Two surgical sites. Design: Retrospective, observational. Methods: Eighty eyes from 73 patients with mild to moderate OAG underwent ab- interno SC viscodilation performed with OMNI system. Fifty eyes (Group 1) underwent only SC viscodilation, while 30 eyes (Group 2) underwent glaucoma surgery + cataract extraction. Primary success endpoint at 12 months was an intraocular pressure (IOP) reduction higher than 25% from baseline with an absolute value of 18 mmHg or lower, either on the same number or fewer ocular hypotensive medications, without further interventions. Secondary effectiveness endpoints included mean IOP, number of medications and comparison of outcomes between groups. Safety endpoints consisted of best-corrected visual acuity (BCVA), adverse events (AEs), and subsequent surgical procedures. Results: Primary success was achieved in 40.0% and 67.9% in Groups 1 and 2, respectively. Mean IOP at 12-month follow-up showed a significant reduction in both groups (from 23.0 to 15.6 mmHg, p < 0.001, and from 21.5 to 14.1, p < 0.001, in Groups 1 and 2, respectively). Mean medication number decreased in both groups (from 3.0 to 2.0, p < 0.001 and from 3.4 to 1.9, p < 0.001, in Groups 1 and 2, respectively). AEs included hyphema (2 eyes), mild hypotony (4 eyes), IOP spikes one month after surgery (1 eye). Twelve eyes (15.0%) required subsequent surgical procedures. No BCVA reduction was observed. Conclusions: Viscodilation of SC using OMNI viscosurgical systems is safe and relatively effective in reducing IOP in adult patients with OAG.

Fourier-Domain Optical Coherence Tomographic Assessment of Changes in the Schlemm’s Canal of Nonglaucomatous Subjects After Keratoplasty

Purpose: This study aimed to evaluate the impact of keratoplasty on the in vivo anatomical structures in the Schlemm’s canal (SC) of nonglaucomatous subjects using Fourier-domain optical coherence tomography (FD-OCT).Methods: Sixty-six nonglaucomatous eyes that underwent penetrating keratoplasty (PK), deep anterior lamellar keratoplasty (DALK), or triple surgery were enrolled in this prospective, comparative, observational study. The SC imaging was performed using FD-OCT before and after surgery in both the nasal and temporal quadrants. Patient demographics, SC parameters [e.g., cross-sectional area (CSA), meridional diameter of SC (MSC), sagittal diameter of SC (SSC), and circumference (CCF)], and the correlations between the variation of SC parameters and intraocular pressure (IOP) were analyzed.Results: The mean age of all subjects was 40.27 ± 18.97 years. Among all cases, the nasal, temporal, and mean MSC significantly decreased on the first day after surgery and then increased at 1 week (p = 0.04, 0.017, and 0.01, respectively). Temporal CSA (tCSA), temporal MSC (tMSC), and temporal circumference (tCCF) after PK (p = 0.017, 0.020, and 0.018, respectively) and nasal MSC (nMSC) after DALK (p = 0.025) decreased significantly on the first day after surgery. The shift in mean IOP was significantly correlated with the changes in tMSC (r = 0.341, p = 0.003) and CCF (r = 0.207, p = 0.048).Conclusion: SC had significant in vivo morphological changes in the early period after keratoplasty in nonglaucomatous eyes, accompanied with elevation of IOP. Early intervention might be necessary to prevent secondary glaucoma early after keratoplasty.

Reduction in trabecular meshwork stem cell content in donor eyes with primary open angle glaucoma

We previously identified and characterized human trabecular meshwork stem cells (TMSCs) based on high expression of ABCG2/p75 positivity and high nucleus to cytoplasmic ratio. These TMSCs expressing high ABCG2 and p75 were located to the insert region of the human TM. Additionally, we demonstrated an age-related reduction in the TMSC content which was significantly associated with TM cell loss. In continuation, this study was aimed to determine the TMSC content in glaucomatous donor eyes wherein a drastic reduction in TM cellularity has already been reported. Anterior segments from known glaucomatous (n = 6) and age-matched normal (n = 8) donors were dissected into four quadrants. A minimum of three sections from each quadrant were used for histopathological analysis as well as immunostaining. Analysis of hematoxylin and eosin-stained sections from glaucomatous tissues revealed a decrease in total TM cellularity, thickening of trabecular beams, fusion of trabeculae, absence of patent Schlemm’s canal compared to age-matched controls. In addition, the TM thickness at various positions of the meshwork and the coronal as well as the meridional diameters of the Schlemm’s canal were observed to be significantly reduced in glaucomatous eyes. Further, sections from both the groups were immunostained for universal stem cell marker ABCG2 and neural crest derived stem cell marker p75. The images were acquired using Leica SP8 confocal microscope. Quantification of total TM cellularity based on nuclear counterstain (mean ± SD) using ImageJ identified 69.33 ± 12.77 cells/section in control eyes. In glaucomatous donors, the TM cellularity was found to be reduced significantly to 41.83 ± 9.0 (p = 0.0007). In addition, a reduction in the percentage of TMSCs (cells with high ABCG2 expression and p75 positivity) was evident in glaucomatous donors (0.14 ± 0.17%) compared to age-matched controls (4.73 ± 5.46%) (p = 0.064). Thus, the present study confirmed the significant decline in TM cellularity and a reducing trend in the TMSC content, though this reduction was non-significant in glaucomatous donor eyes. Further studies are essential to elucidate the role of TMSCs in the pathogenesis of primary open angle glaucoma.

The Relationship Between Age and the Morphology of the Crystalline Lens, Ciliary Muscle, Trabecular Meshwork, and Schlemm’s Canal: An in vivo Swept-Source Optical Coherence Tomography Study

Purpose: To evaluate the effects of age on the morphologies of the crystalline lens, ciliary muscle (CM), Schlemm’s canal (SC), and trabecular meshwork (TM) using swept-source optical coherence tomography (SS-OCT).Methods: Images of the crystalline lens and iridocorneal angle were obtained in healthy participants’ eyes using SS-OCT. Morphological parameters of the crystalline lens, CM, and TM/SC were measured, and the relationship between these parameters and age was evaluated.Results: A total of 62 healthy participants were enrolled, with an age range of 7–79 years. With adjustments for the effects of axial length and sex, both the nasal and temporal SC cross-sectional areas (CSA) and the cross-sectional area of the CM (CMA), distance from the scleral spur to the inner apex of the ciliary muscle (IA-SS), and nasal SC volume were negatively correlated with age (P ≤ 0.041). Meanwhile, the lens thickness (LT) (P &lt; 0.001) and lens vault (LV) (P &lt; 0.001) were positively correlated with age, and the radius of the curvature of the anterior lens (ALR) was negatively correlated with age (P &lt; 0.001).Conclusion: Increasing age was associated with a thicker crystalline lens, a steeper anterior lens curvature, an anteriorly located and smaller CM, and a narrower SC.Clinical Trial Registration:https://register.clinicaltrials.gov/prs/app/action/Select Protocol?sid=S000A3JZ&amp;selectaction=Edit&amp;uid=U00019K7&amp;ts=4&amp;cx=-c5xxp8, identifier [NCT04576884].

The effects of exosomes derived from trabecular meshwork cells on Schlemm’s canal endothelial cells

Trabecular meshwork (TM) and Schlemm’s canal (SC) are the main structures within the conventional outflow pathway, and TM cells and SC endothelial (SCE) cells are essential for controlling intraocular pressure. To examine the interaction between TM cells and SCE cells, we investigated whether exosomes contribute to intercellular communication. Additionally, TM cells in glaucoma acquire mesenchymal characteristics in response to transforming growth factor (TGF)-β2 and extracellular matrix proteins such as collagen type 1 (Col-1); these changes result in increased resistance of aqueous outflow. In this study, we stimulated TM cells with TGF-β2 and Col-1 and characterized the exosomal miRNAs (exomiRs) released in response to each stimulus. Isolated exosomes were rich in miRNAs, with downregulated miR-23a-5p and upregulated miR-3942-5p and miR-7515 levels following Col-1 or TGF-β2 stimulation. Next, a miRNA-mRNA network under TGF-β2 stimulation was constructed. There were no connections among the 3 miRNAs and predicted genes under Col-1 stimulation. GO and KEGG analyses revealed that the identified miRNAs were associated with various signaling pathways, including the inflammatory response. Interestingly, SCE cells treated with miR-7515 mimic showed increased VEGFA, VEGFR2, PECAM, and Tie2 expression. Ultrastructures typical of exosomes and positive staining for exosomal markers were observed in human TM cells. Our data showed that TM cells may communicate with SCE cells via exomiRs and that miR-7515 may be important for SCE cell reprogramming.