Neuropathic Corneal Pain: Tear Proteomic and Neuromediator Profiles, Imaging Features, and Clinical Manifestations.

PURPOSE: To investigate the tear proteomic and neuromediator profiles, in vivo confocal microscopy (IVCM) imaging features, and clinical manifestations in neuropathic corneal pain (NCP) patients. DESIGN: Cross-sectional study. METHODS: A total of 20 NCP patients and 20 age-matched controls were recruited. All subjects were evaluated by corneal sensitivity, Schirmer test, tear break-up time, and corneal and ocular surface staining, Ocular Surface Disease Index and Ocular Pain Assessment Survey questionnaires were administered, as well as IVCM examinations for corneal nerves, microneruomas, and epithelial and dendritic cells. Tears were collected for neuromediator and proteomic analysis using enzyme-linked immunosorbent assay and data-independent acquisition mass spectrometry. RESULTS: Burning and sensitivity to light were the 2 most common symptoms in NCP. A total of 188 significantly dysregulated proteins, such as elevated metallothionein-2, creatine kinases B-type, vesicle-associated membrane protein 2, neurofilament light polypeptide, and myelin basic protein, were identified in the NCP patients. The top 10 dysregulated biological pathways in NCP include neurotoxicity, axonal signaling, wound healing, neutrophil degradation, apoptosis, thrombin signaling mitochondrial dysfunction, and RHOGDI and P70S6K signaling pathways. Compared to controls, the NCP cohort presented with significantly decreased corneal sensitivity (P < .001), decreased corneal nerve fiber length (P = .003), corneal nerve fiber density (P = .006), and nerve fiber fractal dimension (P = .033), as well as increased corneal nerve fiber width (P = .002), increased length, total area and perimeter of microneuromas (P < .001, P < .001, P = .019), smaller corneal epithelial size (P = .017), and higher nerve growth factor level in tears (P = .006). CONCLUSIONS: These clinical manifestations, imaging features, and molecular characterizations would contribute to the diagnostics and potential therapeutic targets for NCP.

Retinal Proteomic Analysis in a Mouse Model of Endotoxin-Induced Uveitis Using Data-Independent Acquisition-Based Mass Spectrometry.

Uveitis is a group of sight-threatening ocular inflammatory diseases, potentially leading to permanent vision loss in patients. However, it remains largely unknown how uveitis causes retinal malfunction and vision loss. Endotoxin-induced uveitis (EIU) in rodents is a good animal model to study uveitis and associated acute retinal inflammation. To understand the pathogenic mechanism of uveitis and screen potential targets for treatment, we analyzed the retinal proteomic profile of the EIU mouse model using a data-independent acquisition-based mass spectrometry (SWATH-MS). After systemic LPS administration, we observed activation of microglial cells accompanied with the elevation of pro-inflammatory mediators and visual function declines. In total, we observed 79 upregulated and 90 downregulated differentially expressed proteins (DEPs). Among the DEPs, we found that histone family members (histone H1, H2A, H2B) and blood proteins including haptoglobin (HP), hemopexin (HPX), and fibrinogen gamma chain (FGG) were dramatically increased in EIU groups relative to those in control groups. We identified phototransduction and synaptic vesicle cycle as the top two significant KEGG pathways. Moreover, canonical pathway analysis on DEPs using Ingenuity Pathway Analysis revealed top three most significant enriched pathways related to acute phase response signaling, synaptogenesis signaling, and eif2 signaling. We further confirmed upregulation of several DEPs associated with the acute phase response signaling including HP, HPX, and FGG in LPS-treated retinas by qPCR and Western blot. In summary, this study serves as the first report to detect retinal proteome changes in the EIU model. The study provides several potential candidates for exploring the mechanism and novel therapeutic targets for uveitis and other retinal inflammatory diseases.

Comparison of A-Scan ultrasonography and the Lenstar optical biometer in Guinea pig eyes.

OBJECTIVE: To compare the biometric parameters provided by A-scan ultrasonography and the Lenstar optical biometer in guinea pig eyes, including anterior segment depth (ASD), lens thickness (LT), vitreous chamber depth (VCD), and axial length (AL), and differences of them between treated form deprivation (FD) eyes and untreated fellow eyes after 4 weeks of FD. METHODS: Three-week-old guinea pigs (N = 41) were subjected to biometric measurements before monocular FD (baseline) and after a 4-week FD. Statistical analyses including within-subject standard deviation (SDwithin), coefficient of variation (CV), and intraclass correlation coefficient (ICC), used to evaluate repeatability for both the A-scan ultrasonography and the Lenstar individually, and correlation and Bland-Altman analyses were used to assess agreement between the two methods. The absolute values of ASD, LT, VCD and AL as measured by the two devices were compared, and the differences of them between treated (T) and untreated fellow (F) eyes (ΔASD, ΔLT, ΔVCD and ΔAL) (Δ = T-F) were compared between the two devices after 4 weeks of FD. RESULTS: Measurements by the Lenstar (ICC: 0.923-0.994) were more repeatable than A-scan ultrasonography (ICC: 0.825-0.870). There was a high correlation for AL (r = 0.851, P < 0.001), a moderate correlation for VCD (r = 0.571, P < 0.001) and LT (r = 0.423, P < 0.001), and a low correlation for ASD (r = 0.230, P < 0.01) between the two devices. The values for ASD, VCD and AL measured by A-scan ultrasonography were larger than those measured by the Lenstar (all, P < 0.001), while LT provided by A-scan ultrasonography was much smaller than that of the Lenstar (P < 0.001). Bland-Altman plots showed poor agreement of absolute values of the four parameters between the two devices. Moreover, there was a high correlation between both methods for ΔAL (r = 0.704, P < 0.001), a moderate correlation for ΔVCD (r = 0.534, P < 0.001) and ΔASD (r = 0.574, P < 0.001), and no correlation for ΔLT (r = 0.303, P = 0.054). The ΔASD, ΔLT, and ΔAL measurements obtained by A-scan ultrasonography were greater than those obtained by the Lenstar (all, P < 0.001), while ΔVCD was mildly smaller using A-scan ultrasonography (P < 0.05). Bland-Altman plots illustrated there is good agreement of ΔAL, ΔVCD, ΔASD, and ΔLT between the two devices. CONCLUSIONS: The Lenstar exhibited better repeatability and provided smaller measurements for AL, VCD and ASD than A-scan ultrasonography. Furthermore, a high correlation and a good agreement for the ΔAL was observed between the two devices after a period of FD. In summary, the two devices cannot replace each other directly to obtain absolute values of ASD, LT, VCD and AL, but the Lenstar still can serve as an option in measuring ΔAL between eyes in guinea pig myopia model.

Combined retinal proteome datasets in response to atropine treatment using iTRAQ and SWATH-MS based proteomics approaches in guinea pig myopia model.

Atropine, a non-selective muscarinic antagonist, is known to slow down myopia progression in human adolescents and in several animal models. However, its underlying molecular mechanism is unclear. The present work built a monocular form-deprivation myopia (FDM) guinea pig model, using facemasks as well as atropine treatment on FDM eyes for 2 and 4 weeks. Retinal protein changes in response to the FDM and effects of topical administration of atropine were screened for the two periods using fractionated isobaric tags for a relative and absolute quantification (iTRAQ) approach coupled with nano-liquid chromatography-tandem mass spectrometry (nano-LC-MS/MS) (n=24, 48 eyes). Retinal tissues from another cohort receiving 4-weeks FDM with atropine treatment (n=12, 24 eyes) with more significant changes were subjected to sequential window acquisition of all theoretical mass spectra (SWATH-MS) proteomics for further protein target confirmation. A total of 1695 proteins (8875 peptides) and 5961 proteins (51871 peptides) were identified using iTRAQ and SWATH approaches, respectively. Using the Paragon algorithm in the ProteinPilotTM software, the three most significantly up-regulated and down-regulated proteins that were commonly found in both ITRAQ and SWATH experiments are presented. All raw data generated from the work were submitted and published in the Peptide Atlas public repository (http://www.peptideatlas.org/) for general release (Data ID PASS01507).

A prospective study of serum metabolomic and lipidomic changes in myopic children and adolescents.

BACKGROUND: Myopia is a prevalent eye disorder, especially among children and adolescents in eastern Asian countries. Multiple measures have already been taken to prevent and treat myopia, including atropine and dopamine. However, the serum metabolic picture of myopia has not yet been studied as a whole and remains largely unclear. In this paper, a prospective and panoramic study was carried out to find out the whole serum metabolomic and lipidomic picture of myopia. METHODS: With untargeted mass spectrometry (MS), myopia among 211 children and adolescents was studied. The MS features were first grouped across the samples. Then, compound annotation was carried out based on these features. Finally, the metabolite features were mapped to pathways, whose biological functions in myopia were studied and discussed. RESULTS: A total of 275 metabolite features were derived from 92 aligned MS peak groups with significant fold changes, and then mapped to 33 pathways. By a comprehensive consideration of significance, fold change, importance score and appearance in different omics, 9 pathways were selected, and their biological functions were further analyzed. Among these selected pathways, 5 pathways were related with oxidative stress, a validated phenomenon during myopia development, while 5 pathways were related with dopamine receptor D2, whose molecular function in myopia treatment is not fully understood. A total of 177 metabolite features from 45 peak groups were related with the studied pathways. CONCLUSION: This prospective study shed light on the whole picture of metabolomic mechanism underlying myopia and provided guidance to further elucidation of compounds and pathways in this whole picture.

Influence of Corneal Topographic Parameters in the Decentration of Orthokeratology.

OBJECTIVE: To investigate the lens decentration (LD) of orthokeratology (ortho-k) and the association between pretreatment corneal topographic parameters and LD of the ortho-k. METHODS: Fifty right eyes of 50 myopes wearing ortho-k lenses were included in the prospective study. Corneal topography was conducted pretreatment to get topographic corneal parameters, including flat-K (K1); steep-K (K2); corneal astigmatism (CA), CA at 0 to 3 mm (3 mm-CA), 3 to 5 mm (5 mm-CA), 5 to 7 mm (7 mm-CA); surface asymmetry index (SAI); surface regularity index; the curvature of best-fit sphere; the diameter of cornea (DC); the distance from the corneal center to the corneal vertex (CCCV); flat eccentricity (E1), steep eccentricity (E2), and E1/E2 (E ratio); and the corneal curvature differences between the nasal and temporal quadrants at 0 to 3 mm (3 mm-Knt), and the corneal curvature differences between the superior and inferior quadrants at 0 to 3 mm (3 mm-Ksi), 5 mm-Knt (at 3-5 mm), 5 mm-Ksi (at 3-5 mm), 7 mm-Knt (at 5-7 mm), and 7 mm-Ksi (at 5-7 mm). The relationship between these cornea topographic parameters and LD of the ortho-k was tested using stepwise multiple linear regression models. RESULTS: The mean magnitude of LD was 0.51±0.23 mm (0.06-1.03 mm). According to the stepwise analysis, 4 factors were associated with the overall LD (P<0.01): SAI (ß=0.252), CCCV (ß=0.539), 5 mm-CA (ß=-0.268), and 3 mm-Ksi (ß=-0.374); 5 factors were associated with the horizontal LD (P<0.01): DC (ß=0.205), CCCV (ß=0.881), 3 mm-CA (ß=-0.217), 5 mm-Knt (ß=0.15), and 3 mm-Ksi (ß=-0.18); and 3 factors were associated with the vertical LD (P<0.01): SAI (ß=0.542), 5 mm-CA (ß=-0.188), and 3 mm-Ksi (ß=-0.213). CONCLUSION: Lens decentration is most common, but in most cases, the amount of LD is moderate and acceptable. The magnitude of LD can be predetermined by topographic corneal parameters. Surface asymmetry index, CCCV, 5 mm-Knt, and 3 mm-Ksi may be more preferable parameters in terms of the assessment of LD of ortho-k.

Efficacy of Trial Fitting and Software Fitting for Orthokeratology Lens: One-Year Follow-Up Study.

OBJECTIVE: To observe and compare the clinical efficacy of 1-year trial fitting and software fitting orthokeratology lenses. METHODS: One hundred myopes who received vision correction with the use of orthokeratology lenses form July 2016 to September 2017 were included in this study. Subjects were assigned randomly into the two groups: the trial fitting group (group A) and the software fitting group (group B). For the right eye of each subject, measurements, such as uncorrected visual acuity (UCVA, logarithm of minimal angle of resolution), refractive error, corneal topography, ocular health status, and the fitting situation, were obtained at baseline, 1 week, 1 month, 3 months, 6 months, and 12 months after lens wear. Axial length and corneal endothelium cells (CECs) were also measured at baseline and 12 months after wearing the lens. RESULTS: Compared with the baseline, the spherical equivalent refraction, UCVA, and central corneal curvature changed significantly after orthokeratology (OK) lens wear (all P<0.05). Between groups A and B, the parameters aforementioned were insignificant at each time point (all P>0.05). Axial length and CECs showed no significant changes during the first year of OK treatment (all P>0.05). Rate of corneal staining between two groups revealed no difference during 1-year visit (P<0.05). CONCLUSION: Both the trial lens fitting and software fitting approaches were effective in temporarily reducing myopia, providing good UCVA and delaying the elongation of axial length for moderate and high myopic adolescents. Both the two approaches can be combined in OK lens fitting.