Sustained descemetocele management with Prosthetic Replacement of the Ocular Surface Ecosystem (PROSE) treatment.

Purpose: To report a case of a 67-year-old male who was successfully managed over a 7-year period for descemetocele secondary to ocular graft versus host disease (oGVHD) using Prosthetic Replacement of the Ocular Surface Ecosystem (PROSE) treatment. Observations: We previously reported on a patient managed with a PROSE device for severe dry eyes secondary to oGVHD, who subsequently developed a central corneal descemetocele. The patient was deemed a poor surgical candidate due to limbal stem cell deficiency secondary to oGVHD. Therefore, we elected to closely monitor the descemetocele as the patient continued PROSE therapy. The patient's descemetocele has been managed successfully without perforation throughout a 7-year follow-up period with corrected distance visual acuity remaining stable at 20/50 in the affected eye. Conclusions and importance: Descemetoceles are an uncommon complication of ocular graft versus host disease. This is the longest published report of a corneal descemetocele managed with PROSE. Our report suggests that in appropriate patients who are at high-risk for post-surgical complications, PROSE in conjunction with other medical management should be considered as an alternative to corneal transplantation.

Spatiotemporal gene targeting in the mouse corneal endothelium.

PURPOSE: The inducible Cre-ERT2 recombinase system allows for temporal control of gene targeting, and it is useful to studying adult function of genes that have critical developmental roles. The Zeb1flox/flox: UBC-CreERT2 mouse was generated to conditionally target Zeb1 to investigate its role in mesenchymal transition in the mouse corneal endothelium in vivo. MATERIALS AND METHODS: Hemizygous UBC-CreERT2 mice were crossed with homozygous mice harboring loxP-flanked Zeb1 alleles (Zeb1flox/flox) to generate the Zeb1flox/flox: UBC-CreERT2 mouse. 4-hydroxytamoxifen (4-OHT) exposure leads to excision of exon 6 of Zeb1, resulting in a loss function allele in the Zeb1flox/flox: UBC-CreERT2 mouse. Intracameral 4-OHT injection further isolates Zeb1 targeting to the anterior chamber. Mesenchymal transition and induction of Zeb1 expression in the corneal endothelium was achieved using FGF2 in ex vivo organ culture. Gene expression was analyzed by semi-quantitative reverse transcription-polymerase chain reaction and by immunoblotting in the mouse corneal endothelium in vivo. RESULTS: Following Cre-mediated targeting of Zeb1 by intracameral 4-OHT injection in Zeb1flox/flox: UBC-CreERT2 mice, FGF2 treatment in ex vivo organ culture resulted in abrogation of Zeb1 mRNA and protein expression in the corneal endothelium. CONCLUSION: The data show Zeb1, a critical mediator of fibrosis in corneal endothelial mesenchymal transition, can be targeted by intracameral injection of 4-OHT in the mouse corneal endothelium in vivo. These results suggest that genes with critical developmental roles can be targeted at a specific time in the corneal endothelium to study its role in adult disease using an inducible Cre-Lox strategy.

Use of Bowman's membrane electrocautery in blind eyes with painful bullous keratopathy not amenable to corneal transplantation: a retrospective case series.

PURPOSE: This study evaluated the safety and efficacy of Bowman's membrane electrocautery in blind painful eyes with bullous keratopathy not amenable to corneal transplantation. METHODS: Eleven eyes of 11 subjects with painful bullous keratopathy and poor visual potential who underwent electrocautery of Bowman's membrane at a tertiary referral ophthalmology clinic were reviewed retrospectively. Subject demographics and preoperative and postoperative data were collected, including description of pain, slit lamp biomicroscopy, best corrected visual acuity, topical medication use, and complications. Efficacy of the procedure on pain reduction, bullae resolution, and topical medication use were assessed at post-operative visits. Safety was also evaluated based on any complications. RESULTS: Bowman's membrane electrocautery effectively resolved bullae in all eyes examined up to 6 months postoperatively; however, 2 eyes had recurrence by 1 year. Mean age at the time of surgery was 69.8 years and mean duration of follow-up was 15.4 months. Pain reduction was achieved in all eyes at 1 month, but 1 subject had pain recurrence by 6 months and another by 1 year. The median number of drops per day decreased from 6 preoperatively to 1.7 at 6 months. Two subjects who had underlying advanced ophthalmic disease had a mild reduction in vision. CONCLUSION: Bowman's membrane electrocautery is a safe and minimally invasive procedure for the management of painful bullous keratopathy in eyes with low vision potential and not amenable to corneal transplantation. Duration of effect appears to last at least 6 months and up to 3 years post-procedure.

Non-infectious and non-hereditary diseases of the corneal epithelium.

The corneal epithelium serves as a physical barrier and a refractive element. Therefore, diseases of the corneal epithelium can increase the risk for infection and causes vision loss. The corneal epithelium can be affected by a multitude of conditions, such as infections, hereditary diseases, depositions, trauma, autoimmune conditions, factitious disorders, and iatrogenic causes. Non-infectious and non-hereditary corneal epithelial diseases represent a collection of conditions with diverse etiologies and clinical presentations but similar patient symptoms. The differing therapeutic interventions for each condition make clinical distinction important. The clinical characteristics, disease course, pathophysiology and current treatments for non-infectious, non-hereditary corneal epithelial diseases are reviewed.

Organogenesis and distribution of the ocular lymphatic vessels in the anterior eye.

Glaucoma surgeries, such as trabeculectomy, are performed to lower intraocular pressure to reduce risk of vision loss. These surgeries create a new passage in the eye that reroutes the aqueous humor outflow to the subconjunctival space, where the fluid is presumably absorbed by the conjunctival lymphatics. Here, we characterized the development and function of the ocular lymphatics using transgenic lymphatic reporter mice and rats. We found that the limbal and conjunctival lymphatic networks are progressively formed from a primary lymphatic vessel that grows from the nasal-side medial canthus region at birth. This primary lymphatic vessel immediately branches out, invades the limbus and conjunctiva, and bidirectionally encircles the cornea. As a result, the distribution of the ocular lymphatics is significantly polarized toward the nasal side, and the limbal lymphatics are directly connected to the conjunctival lymphatics. New lymphatic sprouts are produced mainly from the nasal-side limbal lymphatics, posing the nasal side of the eye as more responsive to fluid drainage and inflammatory stimuli. Consistent with this polarized distribution of the ocular lymphatics, a higher drainage efficiency was observed in the nasal side than the temporal side of the eye when injected with a fluorescent tracer. In contrast, blood vessels are evenly distributed at the anterior surface of the eyes. Also, we found that these distinct vascular distribution patterns were conserved in human eyes. Together, our study demonstrated that the ocular surface lymphatics are more densely present in the nasal side and uncovered the potential clinical benefits in selecting the nasal side as a glaucoma surgery site to improve fluid drainage.

ZEB1 Mediates Fibrosis in Corneal Endothelial Mesenchymal Transition Through SP1 and SP3.

Purpose: ZEB1 is induced during endothelial-mesenchymal transition (EnMT) in the cornea. Induction of SP1 and SP3 by ZEB1 along with identification of putative SP1 and SP3 binding sites in promoters of EnMT-associated gene lead us to investigate their roles in retrocorneal membrane formation in the corneal endothelium. Methods: Expressions of SP1, SP3, and EnMT associated genes were analyzed by immunoblotting and semiquantitative reverse transcription polymerase chain reaction. Accell SMARTpool siRNAs targeting ZEB1, SP1, and SP3 were used for gene knockdown. SP1 and SP3 binding to promoters of EnMT associated genes was investigated by chromatin immunoprecipitation assay. Corneal endothelium in mice was surgically injured in vivo under direct visualization. Results: Transient Fibroblast Growth Factor 2 stimulation increased the expression of both SP1 and SP3 in the human corneal endothelium ex vivo. ZEB1 siRNA knockdown inhibited FGF2-induced SP1 mRNA and protein but not the expression of SP3. FGF2-induced expression of EnMT-related genes, such as fibronectin, vimentin, and type I collagen, was reduced by both SP1 and SP3 siRNA knockdown, with inhibition of SP1 having a greater inhibitory effect than SP3. Additionally, although SP1 and SP3 proteins were found to bind together, SP1 and SP3 could bind to the same promoter binding sites of EnMT-related genes in the absence of the other. Moreover, siRNA knockdown of Zeb1 inhibited injury-dependent RCM formation in mouse corneal endothelium in vivo. Conclusions: Zeb1, through SP1 and SP3, plays a central role in mesenchymal transition induced fibrosis in the corneal endothelium and suggests that Zeb1 could be targeted to inhibit anterior segment fibrosis.

Tear Proteases and Protease Inhibitors: Potential Biomarkers and Disease Drivers in Ocular Surface Disease.

Tears are highly concentrated in proteins relative to other biofluids, and a notable fraction of tear proteins are proteases and protease inhibitors. These components are present in a delicate equilibrium that maintains ocular surface homeostasis in response to physiological and temporal cues. Dysregulation of the activity of protease and protease inhibitors in tears occurs in ocular surface diseases including dry eye and infection, and ocular surface conditions including wound healing after refractive surgery and contact lens (CL) wear. Measurement of these changes can provide general information regarding ocular surface health and, increasingly, has the potential to give specific clues regarding disease diagnosis and guidance for treatment. Here, we review three major categories of tear proteases (matrix metalloproteinases, cathepsins, and plasminogen activators [PAs]) and their endogenous inhibitors (tissue inhibitors of metalloproteinases, cystatins, and PA inhibitors), and the changes in these factors associated with dry eye, infection and allergy, refractive surgery, and CLs. We highlight suggestions for development of these and other protease/protease inhibitor biomarkers in this promising field.

Injury induces endothelial to mesenchymal transition in the mouse corneal endothelium in vivo via FGF2.

Purpose: To determine whether the mouse corneal endothelium enters endothelial to mesenchymal transition (EndoMT) following surgical injury in vivo. Methods: The corneal endothelium in anesthetized mice was surgically injured in vivo under direct visualization. The secretion of interleukin-1 beta (IL-1ß) and fibroblast growth factor 2 (FGF2) into the aqueous humor was analyzed with western blotting. The expression of FGF2, Snai1, Zeb1, Col1a1, Col1a2, Fn1, Vim, Cdk2, Ccne1, and Cdh1 was analyzed with semiquantitative RT-PCR in the mouse corneal endothelium ex vivo and in vivo. Knockdown of FGF2 was done using siRNA. Col8a2 was used as a corneal endothelial marker, and Keratocan (Ktcn) was used as a stromal marker. ß-actin was used as a loading control. Results: Sequential expression of IL-1ß and FGF2 was detected in the aqueous humor after surgical injury. FGF2 treatment induced expression of endothelial to mesenchymal transition-related genes including Snai1, and Zeb1 in the mouse ex vivo corneal endothelium. This led to increased expression of Col1a1, Col1a2, Fn1, and Vim and suppression of Cdh1 in a time-dependent manner. Expression of FGF2, Snai1, Zeb1, Col1a1, Col1a2, Fn1, Vim, Cdk2, and Ccne1 was completely abolished by FGF2 siRNA knockdown in the mouse corneal endothelium ex vivo. Surgical injury induced FGF2 expression in the in vivo mouse corneal endothelium. The injury-dependent expression of FGF2, Snai1, Zeb1, Col1a1, Col1a2, Fn1, Vim, Cdk2, and Ccne1 and the suppression of Cdh1 were inhibited by siRNA knockdown of FGF in the mouse corneal endothelium in vivo. Moreover, siRNA knockdown of FGF2 inhibited the formation of the injury-dependent retrocorneal membrane in the in vivo mouse corneal endothelium. Conclusions: These findings suggest that after surgical injury, FGF2 induces the expression of EndoMT-related genes Snai1, Zeb1, Col1a1, Col1a2, Fn1, Vim, Cdk2, and Ccne1 in the mouse corneal endothelium in vivo, similar to the human corneal endothelium ex vivo.

Ultrasonic Microelastography to Assess Biomechanical Properties of the Cornea.

OBJECTIVE: To both qualitatively and quantitatively investigate corneal biomechanical properties through an ultrasonic microelastography imaging system, which is potentially useful in the diagnosis of diseases, such as keratoconus, postrefractive keratectasia, and tracking treatment such as cross-linking surgery. METHODS: Our imaging system has a dual-frequency configuration, including a 4.5 MHz ring transducer to push the tissue and a confocally aligned 40 MHz needle transducer to track micron-level displacement. Two-dimensional/three-dimensional acoustic radiation force impulse (ARFI) imaging and Young's modulus in the region of interest were performed on ex vivo porcine corneas that were either cross-linked using formalin solution or preloaded with intraocular pressure (IOPs) from 5 to 30 mmHg. RESULTS: The increase of corneal stiffness and the change in cross-linked volume following formalin crosslinking could be precisely observed in the ARFI images and reflected by the reconstructed Young's modulus while the B-mode structural images remained almost unchanged. In addition, the relationship between the stiffness of the cornea and IOPs was investigated among 12 porcine corneas. The corneal stiffness is significantly different at various IOPs and has a tendency to become stiffer with increasing IOP. CONCLUSION: Our results demonstrate the principle of using ultrasonic microelastography techniques to image the biomechanical properties of the cornea. Integrating high-resolution ARFI imaging labeled with reconstructed Young's modulus and structural imaging of the cornea can potentially lead to a routinely performed imaging modality in the field of ophthalmology.

Clusterin from human clinical tear samples: Positive correlation between tear concentration and Schirmer strip test results.

PURPOSE: To investigate the relationship between tear concentration of the homeostatic protein clusterin (CLU) and dry eye signs and symptoms, and to characterize tear CLU protein. METHODS: Two independent studies were conducted, one in Tucson (44 subjects), the other in Los Angeles (52 subjects). A cohort study design was employed to enroll patients without regard to dry eye diagnosis. Dry eye signs and symptoms were assessed using clinical tests. Tear samples were collected by Schirmer strip, and also by micropipette at slit lamp when possible. CLU from both sample types was quantified by immunoassay. The relationship between CLU concentration and clinical test scores was determined by Pearson's correlation coefficient (for individual eyes) and multiple linear regression analysis (including both eyes). CLU was also evaluated biochemically by western blotting. RESULTS: In the Tucson cohort, a positive correlation was observed between tear CLU concentration and results of the Schirmer strip test, a measure of tear flow (p = 0.021 includes both eyes). This result was corroborated in the Los Angeles cohort (p = 0.013). The mean tear CLU concentration was 31 ±â€¯14 µg/mL (n = 18 subjects, 33 eyes; range = 7-48 µg/mL). CLU from clinical tear samples appeared biochemically similar to CLU from a non-clinical tear sample and from blood plasma. CONCLUSIONS: Results support the hypothesis that an optimal concentration of tear CLU is important for ocular surface health, and that this drops below the effective threshold in dry eye. Tear CLU measurement might identify patients that could benefit from supplementation. Information about concentration will aid development of therapeutic dosage parameters.

Increased Cathepsin S activity associated with decreased protease inhibitory capacity contributes to altered tear proteins in Sjögren's Syndrome patients.

Cathepsin S (CTSS) activity is elevated in Sjögren's Syndrome (SS) patient tears. Here we tested whether protease inhibition and cystatin C (Cys C) levels are reduced in SS tears, which could lead to enhanced CTSS-driven degradation of tear proteins. CTSS activity against Cys C, LF and sIgA was tested in SS or healthy control tears. Tears from 156 female subjects (33, SS; 33, rheumatoid arthritis; 31, other autoimmune diseases; 35, non-autoimmune dry eye (DE); 24, healthy controls) were analyzed for CTSS activity and Cys C, LF, and sIgA levels. Cys C and LF showed enhanced degradation in SS tears supplemented with recombinant CTSS, but not supplemented healthy control tears. CTSS activity was significantly increased, while Cys C, LF and sIgA levels were significantly decreased, in SS tears compared to other groups. While tear CTSS activity remained the strongest discriminator of SS in autoimmune populations, combining LF and CTSS improved discrimination of SS beyond CTSS in DE patients. Reductions in Cys C and other endogenous proteases may enhance CTSS activity in SS tears. Tear CTSS activity is reconfirmed as a putative biomarker of SS in an independent patient cohort while combined LF and CTSS measurements may distinguish SS from DE patients.

Fibroblast growth factor 2 induces proliferation and fibrosis via SNAI1-mediated activation of CDK2 and ZEB1 in corneal endothelium.

Investigating stimulation of endogenous wound healing in corneal endothelial cells (CECs) may help address the global shortage of donor corneas by decreasing the number of transplants performed for blindness because of endothelial dysfunction. We previously reported that IL-1ß stimulation leads to fibroblast growth factor (FGF2) expression, enhancing migration and proliferation of mammalian CECs. However, FGF2 also promotes the endothelial-mesenchymal transition, which can lead to retrocorneal membrane formation and blindness. This prompted us to investigate downstream FGF2 signaling targets that could be manipulated to prevent retrocorneal membrane formation. FGF2 stimulation altered cell morphology and induced expression of mesenchymal transition marker genes such as snail family transcriptional repressor 1 (SNAI1), SNAI2, zinc finger E-box-binding homeobox 1 (ZEB1), and ZEB2 This, in turn, induced expression of fibronectin, vimentin, and type I collagen, and suppressed E-cadherin in CECs in vitro and ex vivo siRNA-mediated SNAI1 knockdown revealed that SNAI1 induces ZEB1 expression, in turn inducing expression of type I collagen, the major component of retrocorneal membranes, and of cyclin-dependent kinase 2 (CDK2) and cyclin E1, promoting cell proliferation. siRNA-mediated knockdown of SNAI1 or ZEB1, but not of CDK2, inhibited FGF2-dependent expression of fibronectin, vimentin, and type I collagen and of suppression of E-cadherin expression. We conclude that SNAI1 is a key regulator of FGF2-dependent mesenchymal transition in human ex vivo corneal endothelium, with ZEB1 regulating type I collagen expression and CDK2 regulating cell proliferation. These results suggest that SNAI1 promotes fibrosis and cell proliferation in human corneal endothelium through ZEB1 and CDK2.

Prosthetic Replacement of the Ocular Surface Ecosystem Scleral Lens Therapy for Exposure Keratopathy.

OBJECTIVES: To investigate the utility of Prosthetic Replacement of the Ocular Surface Ecosystem (PROSE) scleral lenses in patients with exposure keratopathy, with outcomes based on changes in visual acuity, visual function, and corneal staining. METHODS: A retrospective interventional case series of patients evaluated and treated from September 2009 through June 2014 at a single multi-specialty institutional practice. Eighteen of 29 patients with exposure keratoconjunctivitis, lagophthalmos, ectropion, or lid retraction, referred to USC Eye Institute after failing conventional therapies completed PROSE scleral lens fitting and were included in the study. Visual function was assessed before and after PROSE fitting with the Ocular Surface Disease Index (OSDI) survey. Visual acuity (VA) and corneal staining changes were also evaluated before and after treatment. RESULTS: Visual acuity improved from 0.60±0.68 logMAR pre-PROSE to 0.25±0.34 logMAR (Z=-3.81, P=0.00014) post-PROSE, which corresponds to an improvement of about 20/80 to 20/35 on Snellen VA. Ocular Surface Disease Index scores improved from 56.54±29.75 pre-PROSE to 24.98±21.23 post-PROSE (Z=-2.98, P=0.0029), and corneal staining values decreased from 2.17±0.84 pre-PROSE to 0.64±0.70 post-PROSE (Z=-3.27, P=0.011). CONCLUSIONS: The results suggest that PROSE scleral lens therapy is effective in patients with exposure keratopathy who had failed conventional therapies and can serve as an alternative to lid surgery.

Clusterin Seals the Ocular Surface Barrier in Mouse Dry Eye.

Dry eye is a common disorder caused by inadequate hydration of the ocular surface that results in disruption of barrier function. The homeostatic protein clusterin (CLU) is prominent at fluid-tissue interfaces throughout the body. CLU levels are reduced at the ocular surface in human inflammatory disorders that manifest as severe dry eye, as well as in a preclinical mouse model for desiccating stress that mimics dry eye. Using this mouse model, we show here that CLU prevents and ameliorates ocular surface barrier disruption by a remarkable sealing mechanism dependent on attainment of a critical all-or-none concentration. When the CLU level drops below the critical all-or-none threshold, the barrier becomes vulnerable to desiccating stress. CLU binds selectively to the ocular surface subjected to desiccating stress in vivo, and in vitro to the galectin LGALS3, a key barrier component. Positioned in this way, CLU not only physically seals the ocular surface barrier, but it also protects the barrier cells and prevents further damage to barrier structure. These findings define a fundamentally new mechanism for ocular surface protection and suggest CLU as a biotherapeutic for dry eye.

WNT10B enhances proliferation through ß-catenin and RAC1 GTPase in human corneal endothelial cells.

The cornea is the anterior, transparent tissue of the human eye that serves as its main refractive element. Corneal endothelial cells are arranged as a monolayer on the posterior surface of the cornea and function as a pump to counteract the leakiness of its basement membrane. Maintaining the cornea in a slightly dehydrated state is critical for the maintenance of corneal transparency. Adult human corneal endothelial cells are G1-arrested, even in response to injury, leading to an age-dependent decline in endothelial cell density. Corneal edema and subsequent vision loss ensues when endothelial cell density decreases below a critical threshold. Vision loss secondary to corneal endothelial dysfunction is a common indication for transplantation in developed nations. An impending increase in demand for and a current global shortage of donor corneas will necessitate the development of treatments for vision loss because of endothelial dysfunction that do not rely on donor corneas. Wnt ligands regulate many critical cellular functions, such as proliferation, making them attractive candidates for modulation in corneal endothelial dysfunction. We show that WNT10B causes nuclear transport and binding of RAC1 and ß-catenin in human corneal endothelial cells, leading to the activation of Cyclin D1 expression and proliferation. Our findings indicate that WNT10B promotes proliferation in human corneal endothelial cells by simultaneously utilizing both ß-catenin-dependent and -independent pathways and suggest that its modulation could be used to treat vision loss secondary to corneal endothelial dysfunction.

Prosthetic Replacement of the Ocular Surface Ecosystem (PROSE) scleral lens for Salzmann's nodular degeneration.

PURPOSE: To evaluate the use of Prosthetic Replacement of the Ocular Surface Ecosystem (PROSE) scleral lens treatment as an alternative to keratectomy in patients with symptomatic Salzmann's nodular degeneration (SND). METHODS: A retrospective chart review from July 2009 to May 2013 identified 9 SND patients who were referred for PROSE evaluation. Patients who did not complete PROSE fitting or had other corneal comorbidities affecting vision were excluded from the study, and 7 eyes of 4 patients were included. Three patients were pseudophakic and 1 patient was phakic, and the lens status of our cohort did not change during the study. RESULTS: Visual acuity improved from 0.19 ± 0.084 logMAR (approximately 20/31) pre PROSE to 0.028 ± 0.047 logMAR (approximately 20/21) post PROSE in patients with Salzmann's nodular degeneration (p = 0.002). OSDI scores improved from 46.9 ± 26.6 pre PROSE to 21.5 ± 18.7 post PROSE in the same cohort (p = 0.02). CONCLUSION: The results of this study show that PROSE can provide improvements in visual acuity and function in patients with Salzmann's nodular degeneration and offer an alternative to superficial keratectomy.

Ocular surface rehabilitation: Application of human amniotic membrane in high-risk penetrating keratoplasties.

BACKGROUND: Human amniotic membrane is a versatile tool for management of ocular surface disorders. This study evaluates the effect of cryopreserved human amniotic membrane (hAM) on one-year survival of penetrating keratoplasties (PKP) in high-risk recipients. METHOD: This is a retrospective noncomparative cohort study of 58 consecutive eyes undergoing PKP with concurrent placement of a self-retained cryopreserved hAM (PROKERA®) at a tertiary care center from January 2009 to July 2010. RESULTS: Mean patient age was 66.7 ± 17.2 years and 30 (54%) were males. 51 eyes were pseudophakic and one aphakic. 27 eyes were glaucomatous; 24 had glaucoma drainage device and 2 had previous endocyclophotocoagulation. 12 patients had PKP for the first time and 46 had repeat PKP (average number of prior PKP = 1.63 ± 1.1, range: 1-5). Risk factors for graft failure included repeat PKP (79.3%), corneal neovascularization (51.7%), preexisting glaucoma (46.6%), and presence of anterior synechiae (37.9%). Both First Transplant and Repeat Transplant groups had similar survival rates until 6 months after transplant (75% vs 74%, odds ratio = 1.06, p = 1.00). At 12 months, First Transplant group showed a better survival rate (67% vs 43%, odds ratio = 2.60, p = 0.20). Eyes with >3 risk factors had a higher graft failure rate (odds ratio = 5.81, p = 0.003). CONCLUSION: Survey of the literature suggests that high-risk PKP with concurrent hAM placement demonstrate comparable graft survival. Presence of multiple risk factors is associated with poor survival.

Interleukin-1ß-induced Wnt5a enhances human corneal endothelial cell migration through regulation of Cdc42 and RhoA.

Wnt5a can activate ß-catenin-independent pathways for regulation of various cellular functions, such as migration, that play critical roles in wound repair. Investigation of Wnt5a signaling may help identify therapeutic targets for enhancing corneal endothelial wound healing that could provide an alternative to corneal transplantation in patients with blindness from endothelial dysfunction. However, Wnt5a signaling in corneal endothelial cells (CECs) has not been well characterized. In this study, we show transient induction of Wnt5a by interleukin-1ß (IL-1ß) stimulation proceeds through NF-κB in human CECs. This leads to binding of Fzd5 to Ror2, resulting in activation of disheveled protein (Dvl) and subsequently disheveled-associated activator of morphogenesis 1 (DAAM1). This leads to activation of Cdc42 and subsequent inhibition of RhoA. Inhibition of RhoA leads to parallel dephosphorylation and inactivation of LIM domain kinase 2 along with dephosphorylation and activation of slingshot 1, resulting in dephosphorylation and activation of cofilin and leading to enhanced cell migration. These findings suggest that Wnt5a enhances cell migration through activation of Cdc42 and inactivation of RhoA in human CECs.

Prosthetic replacement of the ocular surface ecosystem scleral lens therapy for patients with ocular symptoms of chronic Stevens-Johnson syndrome.

PURPOSE: To evaluate the results of Prosthetic Replacement of the Ocular Surface Ecosystem (PROSE) scleral lens treatment on visual acuity and function in patients with ocular symptoms of chronic Stevens-Johnson syndrome (SJS). DESIGN: Retrospective interventional case series. METHODS: setting: Single multi-specialty institutional practice. study population: A chart review from July 2009 to July 2013 identified 19 patients with ocular symptoms from chronic SJS who were referred for PROSE fitting evaluation. Three patients deemed appropriate candidates were excluded because they were lost to follow-up during the fitting process. Only 1 eye was fitted in 4 patients because anatomic changes prohibited PROSE fitting in the fellow eye. Another patient chose to have PROSE fitting only in 1 eye. A total of 27 eyes of 16 patients who completed PROSE fitting were included in this study. intervention: PROSE scleral lens fitting. outcome measures: Visual acuity and visual function were assessed before and after PROSE fitting using Snellen acuity and Ocular Surface Disease Index (OSDI) survey. The OSDI survey is a validated questionnaire that assesses ocular surface disease in the context of vision-related function, ocular symptoms, and environmental triggers. RESULTS: Visual acuity improved from 0.43 ± 0.35 logMAR pre-PROSE to 0.14 ± 0.22 logMAR post-PROSE (P = .0007) in SJS patients. OSDI scores improved from 70.4 ± 19.0 pre-PROSE to 37.4 ± 23.2 post-PROSE (P = .0002) in the same cohort. CONCLUSION: The results of this study show that PROSE treatment is a viable option for improving visual acuity and function in SJS patients who failed conventional treatment.

Tear cathepsin S as a candidate biomarker for Sjögren's syndrome.

OBJECTIVE: The diagnosis of Sjögren's syndrome (SS) in routine practice is largely a clinical one and requires a high index of suspicion by the treating physician. This great dependence on clinical judgment frequently leads to delayed diagnosis or misdiagnosis. Tear protein profiles have been proposed as simple and reliable biomarkers for the diagnosis of SS. Given that cathepsin S activity is increased in the lacrimal glands and tears of NOD mice (a murine model of SS), the aim of this study was to explore the clinical utility of using tear cathepsin S (CTSS) activity as a biomarker for SS. METHODS: A method to measure CTSS activity in tears eluted from Schirmer's test strips was developed and validated. Schirmer's tests were performed and CTSS activity measurements were obtained in 278 female subjects, including 73 with SS, 79 with rheumatoid arthritis, 40 with systemic lupus erythematosus, 10 with blepharitis, 31 with nonspecific dry eye disease, and 12 with other autoimmune diseases, as well as 33 healthy control subjects. RESULTS: The median tear CTSS activity in patients with SS was 4.1-fold higher than that in patients with other autoimmune diseases, 2.1-fold higher than that in patients with nonspecific dry eye disease, and 41.1-fold higher than that in healthy control subjects. Tear CTSS levels were equally elevated in patients with primary SS and those with secondary SS, independent of the Schirmer's test strip values or the levels of circulating anti-SSA or anti-SSB antibodies. CONCLUSION: Markedly high levels of tear CTSS activity are suggestive of SS. CTSS activity in tears can be measured in a simple, quick, economical, and noninvasive manner and may serve as a novel biomarker for autoimmune dacryoadenitis during the diagnostic evaluation for SS.