MicroRNA profiling in the aqueous humor of highly myopic eyes by next generation sequencing
ABSTRACT: Purpose: To characterize microRNAs (miRNAs) and their possible roles in high myopia by using next generation sequencing Methods: Aqueous humor samples were obtained from 15 highly myopic eyes and 15 cataract eyes at the onset of surgery. miRNA next generation sequencing and bioinformatics analyses were performed using RNA extracted from aqueous humor samples. Results: A total of 341 miRNAs were detected in the aqueous humor samples of highly myopic eyes; 201 miRNAs were detected in the aqueous humor samples of cataractous control eyes. A total of 249 mature miRNAs and 17 novel miRNAs were differentially expressed during myopia. Possible pathways regulated by these aberrantly expressed miRNAs included the TNF, MAPK, PI3K-Akt, and HIF-1 signaling pathways. Conclusions: The current study provided an overall view of miRNA profiling in the aqueous humor of highly myopic eyes. These profiles may be associated with myopia pathogenesis, and are potential biomarkers. Overall design: miRNA profiles in aqueous humor of highly myopic patients (15 eyes of 15 patients), aqueous humor of cataract patients as control (15 eyes of 15 patients) , aquous humor from 3 eyes mixed as 1 sample, detected in quintuplicate, using Illumina Hiseq4000
Project description:The study aim was to determine microRNA (miRNA) expression profiles of ocular tissues in form-deprivation induced myopia (FDM) mice. Form-deprivation myopia was induced in C57BL/6Jmice over the right eye; the contralateral left eyes were used as controls. Whole genome microRNA expression profiles in myopic whole eye, retina, and sclera were determined using the Agilent mouse miRNA microarray. The normalized microarray data were performed ANOVA test to identify differences in miRNA expression between myopic and control eyes. The differential expression for selected miRNAs was validated by quantitative real time PCR (qRT-PCR). Overall design: Fifteen C57BL/6J mice were used in this study. Form-deprivation myopia was induced over the right eyes; the contralateral left eyes were used as controls. Total RNA was isolated from the whole eye (6 mice), retina (9 mice), and sclera (9 mice) and was used for miRNA expression profiling with the Agilent Mouse miRNA v.15 Microarray.
Project description:<h4>Purpose</h4>To evaluate the deeper structures of the optic nerve and to analyze the shape of eyes with tilted disc syndrome (TDS) by swept-source optical coherence tomography (OCT) and three-dimensional magnetic resonance imaging (3D MRI).<h4>Methods</h4>The medical records of 54 eyes of 36 patients with TDS were reviewed. The patients with TDS and high myopia were analyzed separately from those without high myopia. All the eyes were examined with a swept-source OCT, and 22 of the eyes were examined by 3D MRI.<h4>Results</h4>A total of 38 eyes of 29 patients were highly myopic and 16 eyes of 15 patients were not highly myopic. The representative OCT findings of the optic disc were: a sloping of the lamina cribrosa posteriorly from the upper part to the lower part, a protrusion of the upper edge of Bruch's membrane, and choroid. The distance and the depth of the most protruded point from the fovea were significantly greater in the eyes with non-highly myopic TDS than those with highly myopic TDS. In the 3D MRI, the lower part of the posterior segment was protruded outward, and the optic nerves attached at the upper nasal edge of the protrusion.<h4>Conclusions</h4>The abnormalities detected by swept-source OCT and 3D MRI analyses indicate the possibility that the essential pathology of TDS is a deformity of the inferior globe below the optic nerve, and the positional relation between the fovea and the inferior protrusion determines the degree of myopia.
Project description:To evaluate the corneal spherical aberrations in cataract patients with and without high myopia, we conducted a retrospective case series of 502 cataract eyes with high myopia and 1500 age-related cataract eyes and measure their corneal biometric data and axial length using Pentacam and IOLMaster. Both the anterior and total corneal primary spherical aberrations were lower in the high myopia group than that in the control group (0.317?±?0.215 vs 0.338?±?0.148 ?m, P?=?0.043; and 0.281?±?0.207 vs 0.314?±?0.153 ?m, P?<?0.001). The incidence of eyes with negative total corneal primary spherical aberration increased as axial length increased in the high myopia group, and the overall incidence was higher in the high myopia group than that in the control group (2.59% vs 1.47%). These were mainly contributed to the younger age of cataract patients with high myopia (55.76?±?13.10 vs 60.18?±?15.72 years, P?<?0.001), along with the positive correlations between age and anterior and total corneal primary spherical aberrations. In clinical practice, an aspheric IOL with a low negative or zero primary spherical aberration is recommended for cataract patients with high myopia. Negative total ocular primary spherical aberrations resulting from aspheric IOL implantation should be avoided in extremely high myopic eyes.
Project description:High myopia is recognized as a risk factor for earlier onset of nuclear cataracts. One possible explanation for this is that lenses in highly myopic eyes are exposed to higher levels of oxygen than normal eyes owing to earlier vitreous liquefaction and, hence, are subjected to oxidative insults. Here, we first compared the methylation levels of six essential antioxidant genes (GSTP1, NRF2, OGG1, TXN, TXNRD1 and TXNRD2) between highly myopic cataract (HMC) and age-related cataract (ARC) lens epithelial samples via Sequenom MassARRAY. We found that specific CpG units in the promoters of GSTP1 and TXNRD2 were hypermethylated and that the expression levels of these two genes were lower in the HMC group than in the ARC group. A luciferase reporter assay confirmed the significance of differentially methylated fragments in the activation of transcription. The importance of GSTP1 and TXNRD2 in antioxidant capacity was confirmed by overexpression or knockdown experiments on cultured lens epithelial cells (LECs). In addition, the expression of DNA methyl transferase 1 (DNMT1) was higher in the lens epithelium of HMC patients than that of ARC patients, and the expression of GSTP1 and TXNRD2 was upregulated by use of a DNMT inhibitor in cultured LECs. Finally, we mimicked the intraocular environment of highly myopic eyes by treating LECs with hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) and observed both alterations in the methylation status of the GSTP1 and TXNRD2 promoters and time-dependent altered expression levels. Therefore, we propose that in an environment with high oxygen, in which lenses in highly myopic eyes are immersed, there exists a vicious cycle composed of increased oxidative stress and decreased enzymatic antioxidants via the hypermethylation of antioxidant genes.
Project description:Background:To measure the crystalline lens tilt in eyes with various degrees of myopia before cataract surgery using swept-source optical coherence tomography (SS-OCT). Methods:We used SS-OCT (IOLMaster 700) to scan 131 emmetropic eyes (axial length?<?24.5?mm), 25 mild/moderate myopic eyes (axial length 24.5-26?mm), and 123 high myopic eyes (52, 29, and 42 eyes with axial lengths of 26-28, 28-30, and?>?30?mm, respectively) as part of the routine preoperative examination before cataract surgery. SS-OCT involved B-scans along six meridians. The data were analyzed to assess the magnitude and orientation of the lens tilt and their correlation with other optical biometric parameters. Result:The mean tilt was 3.36?±?0.98° in emmetropic eyes, 3.07?±?1.04° in mild/medium myopic eyes, and 2.35?±?1.01° in high myopic eyes. Tilt correlated significantly and inversely with axial length (Pearson's r?=?-?0.427, P?<?0.001). The crystalline lens tilt predominantly faced the upper outer quadrant relative to the visual axis, symmetrically in both eyes, with mean angles of 24.32° and 147.36° in the right and left eyes, respectively. The variability in the lens tilt direction increased with increasing axial length (?2 test, P?<?0.001). Conclusion:The magnitude of crystalline lens tilt decreased with increasing axial length. The direction of tilt was predominantly towards the upper outer quadrant in both eyes. The variability in the tilt orientation increased with increasing axial length. Trial registration:NIH (clinicaltrial.gov), NCT03062085. Registered 23 February 2017.
Project description:Inflammation is known to be involved in the progression of diabetic retinopathy. We have recently reported that vitreous levels of IL-4, IL-17A, IL-22, IL-31, and TNF? are higher than the respective serum levels in proliferative diabetic retinopathy (PDR) patients, and that vitreous levels of these cytokines are higher in PDR than in other non-inflammatory vitreoretinal diseases or uveitis associated with sarcoidosis. In the present study, we investigated inflammatory cytokines including Th17 cell-related cytokines in aqueous humor samples obtained from eyes with PDR, and analyzed the association between the aqueous humor and vitreous fluid levels of individual cytokines. The study group consisted of 31 consecutive type 2 diabetic patients with PDR who underwent cataract surgery and vitrectomy for vitreous hemorrhage and/or tractional retinal detachment. Undiluted aqueous humor was collected during cataract surgery, and then vitreous fluid was obtained using a 25G vitreous cutter inserted into the mid-vitreous cavity at the beginning of vitrectomy. IL-1?, IL-4, IL-6, IL-10, IL-17A, IL-17F, IL-21, IL-22, IL-23, IL-25, IL-31, IL-33, IFN-?, soluble CD40 ligand (sCD40L), and TNF? levels in the aqueous humor and vitreous fluid were measured using a beads-array system. Although IL-17A was detected in the aqueous humor of eyes with PDR and the level correlated with IL-17A level in the vitreous fluid, both percent detectable and level of IL-17A in the aqueous humor were significantly lower than those in the vitreous fluid. Vitreous IL-17A level was related significantly to IL-10, IL-22, and TNF? levels in aqueous humor as well as in vitreous fluid, On the other hand, aqueous IL-17A level was not related significantly to aqueous or vitreous levels of IL-10, IL-22 or TNF? level. The present study demonstrated that IL-17A level and detectable rate in the aqueous humor of patients with PDR are markedly lower than those in the vitreous fluid and aqueous IL-17A does not correlate with vitreous levels of other cytokines, and hence should not be used as a surrogate for IL-17A in the vitreous fluid.
Project description:Purpose:To segment and quantify three distinct retinal capillary plexuses using optical coherence tomography angiography (OCTA) in myopic eyes. Methods:We analyzed 96 eyes from 62 subjects with myopia (27.76 ± 7.05 years of age) and evaluated 30 normal eyes from 15 subjects (28.33 ± 3.13 years of age) for controls. En face OCTA images generated by AngioPlex (Carl Zeiss; Oberkochen, Germany) were manually segmented by the progressive matching method into superficial, middle, and deep capillary plexuses (SCPs, MCPs, and DCPs, respectively). Estimated positions for each plexus relative to the reference line were calculated. After strict artifact removal and magnification correction, vessel density (VD) and skeleton density (SD) analyses were performed on each capillary plexus. Results:Myopic eyes were divided into three groups according to their degree of myopia. We defined the relative estimated positions of the MCP outer boundary to the retinal pigment epithelium fit layer as MCP = -89.317 - 0.178 (central retinal thickness) - 0.580 (ganglion cell inner plexiform thickness); the DCP outer boundary was 38.48 ± 6.24 µm below the inner plexiform layer. VDs were significantly higher in the super-high myopia group than in the control and moderate myopia groups for the DCP (all P < 0.05). SDs in the SCPs were significantly lower in the high myopia and super-high myopia groups than in the control groups (all P < 0.001). Conclusions:With progressive matching, we segmented three capillary plexuses and defined the relative estimated positions of each capillary plexus to the reference line in myopic eyes. The VD of the DCP increased for more myopic eyes. Translational Relevance:Our study provides a visual method for OCTA image vascular segmentation for myopic eyes.
Project description:To describe cytokines, chemokines and growth factors profiles in patients undergoing cataract surgery with femtosecond laser pretreatment and investigate their relationships with the postoperative in vivo inflammation index.Aqueous humor was collected from 22 eyes after femtosecond laser pretreatment and from 22 eyes at the beginning of routine cataract surgery. The levels of 45 inflammation-related mediators were measured using multiplex fluorescent bead-based immunoassays. Laser flare photometry was measured preoperatively and at 1 day, 7 days and 30 days postoperatively.Compared with the control group, the femtosecond laser treatment group showed significantly higher aqueous humor levels of fibroblast growth factor (FGF-2), tumor necrosis factor (TNF)-α, leukemia inhibitor factor (LIF), interleukin (IL)-1ra and IL-18, and significantly lower aqueous humor levels of IL-9, platelet-derived growth factor (PDGF)-BB, eotaxin and TNF-β. Postoperative aqueous flare was significantly greater in the manual cataract surgery group at 1 day (p<0.001), 7 days (p<0.001) and 30 days (p = 0.002).No correlation was found between the analyzed mediators and the aqueous flare values.The expression profiles of cytokines, chemokines and growth factors and the correlations of these profiles with the in vivo inflammatory indexes for patients undergoing cataract surgery with femtosecond laser pretreatment were identified. Our data indicate a disturbance of postoperative inflammation response after femtosecond laser treatment.
Project description:Purpose:To investigate the change in cytokine microenvironment of the aqueous humor (AH) after surgery in children with congenital or developmental cataracts. Methods:AH samples were obtained from 59 eyes diagnosed with a congenital or developmental cataract. Thirty-three of these eyes were aphakic following previous cataract surgery and were scheduled for secondary intraocular lens (IOL) implantation. Additionally, AH samples from 26 eyes that had not undergone surgery were taken. AH samples were then analyzed for 16 different inflammatory immune mediators using multiplex bead immunoassays and enzyme-linked immunosorbent assay (ELISA). Results:The mean interval between secondary IOL implantation and original cataract surgery was 24.85 months (range, 9-60 months). Levels of IL-6, IP-10 (CXCL10), MCP-1 (CCL2), and IL-2 were significantly elevated in the AH of eyes after surgery compared to eyes that did not undergo surgery (P < 0.001, P = 0.047, P = 0.006, P = 0.012, respectively). There was significant correlation between the levels of TGF-?2 and intraocular pressure (IOP) in postsurgical and nonsurgical eyes (r = 0.532, P = 0.006; r = 0.57, P = 0.001). Postsurgical outcomes, such as iris adhesions, capsular fibrosis, and capsular contraction, were found not to be significantly associated with cytokine levels in the AH after surgery; however, IL-6 levels in capsular exposure eyes were significantly higher than those in cortical closure eyes (P = 0.023). Conclusions:To our knowledge, this is the first study to report significantly increased proinflammatory cytokine levels in the AH after congenital cataract extraction in children. Our study also suggests that this proinflammatory state may be maintained for a prolonged period of time. Overall, these results give us insight into the relationship between the inflammatory cytokine microenvironment of the aqueous humor and potential long-term complications following congenital cataract surgery. Translational Relevance:The inflammatory cytokine microenvironment of the aqueous humor might help explain potential long-term complications after surgery in patients with congenital or developmental cataracts.