Next-generation sequencing-based gene panel tests for the detection of rare variants and hypomorphic alleles associated with primary open-angle glaucoma.

Primary open-angle glaucoma (POAG) is a complex disease with a strong hereditably component. Several genetic variants have recently been associated with POAG, partially due to technological improvements such as next-generation sequencing (NGS). The aim of this study was to genetically analyze patients with POAG to determine the contribution of rare variants and hypomorphic alleles associated with glaucoma as a future method of diagnosis and early treatment. Seventy-two genes potentially associated with adult glaucoma were studied in 61 patients with POAG. Additionally, we sequenced the coding sequence of CYP1B1 gene in 13 independent patients to deep analyze the potential association of hypomorphic CYP1B1 alleles in the pathogenesis of POAG. We detected nine rare variants in 16% of POAG patients studied by NGS. Those rare variants are located in CYP1B1, SIX6, CARD10, MFN1, OPTC, OPTN, and WDR36 glaucoma-related genes. Hypomorphic variants in CYP1B1 and SIX6 genes have been identified in 8% of the total POAG patient assessed. Our findings suggest that NGS could be a valuable tool to clarify the impact of genetic component on adult glaucoma. However, in order to demonstrate the contribution of these rare variants and hypomorphic alleles to glaucoma, segregation and functional studies would be necessary. The identification of new variants and hypomorphic alleles in glaucoma patients will help to configure the genetic identity of these patients, in order to make an early and precise molecular diagnosis.

Risk Factors for Failure in Double-Plate Tube Surgery for Refractory Glaucoma: 25 Years Surgical Experience.

PURPOSE: To investigate risk factors associated with success and failure in double-plate tube surgery. METHODS: This retrospective case-series observational study included 243 consecutive eyes that underwent anterior-segment double-plate tube surgery from 1990 to 2015. Evaluation of the efficacy of the device was based on the final intraocular pressure (IOP) and the need for anti-glaucoma medication. We also assessed success and failure according to risk factors for trabeculectomy and an early hypertensive phase (HP). RESULTS: Preoperative IOP was 37.3±13.1 mmHg (mean±SD) with 3.0±0.7 medications. After a median follow-up of 44.3 months, the mean IOP was 14.6±6.3 mmHg with 0.4±1.0 medications. The final IOPs ranged from 6 to 21 mmHg in 87.24% of eyes; however, 25.47% required medication. No risk factors studied were associated with surgical failure. Preoperative IOP, glaucoma type, previous surgery, previous anti-glaucoma drugs, implant type, and HP were associated with partial success (p<0.05). HP and preoperative use of brimonidine reduced the probability of complete success by 66.9% and 68.2%, respectively (p<0.05). HP was more likely when chronic preoperative prostaglandin analogues were administered (odds ratio [OR] 4.286; 95% confidence intervals [CI] 1.593-11.529; P=0.0039) and when the tube was located in the posterior chamber (OR 3.561; 95% CI 1.286-9.861; P=0.0145). CONCLUSION: Tube surgery is effective and seems to be independent of the major risk factors for glaucoma surgery. However, previous surgery and some chronic preoperative drugs are related to the need for glaucoma medication to achieve the target pressure.

Hemorrhagic Descemet's Membrane Detachment in Nonpenetrating Glaucoma Surgery.

PURPOSE: To describe 5 representative cases of hemorrhagic Descemet's membrane (DM) detachment in glaucoma surgery that had different origins, mechanisms and treatments. METHODS: Clinical records of patients that had undergone a nonpenetrating glaucoma surgery, with a diagnosis of hemorrhagic DM detachment were reviewed for demographic data, clinical findings and treatment applied. RESULTS: Five patients with hemorrhagic DM detachment were included in this case series. They all had different causes, namely a massive hemorrhage at the end of a canaloplasty procedure, a needling maneuver, autologous blood injection, Swan syndrome, and frequent eye rubbing. Hematoma evacuation was performed in 4 eyes, 1 of them from under the scleral flap of the deep sclerectomy and 3 of them through a surgical or laser perforation in DM. Air tamponade was done in most of these cases. One of these cases required transcorneal suture fixation. One case was observed expectantly. All cases successfully recovered but peripheral corneal stain was persistent in 2 cases. CONCLUSION: Hemorrhagic DM detachment is a rare but potential vision-threatening complication in glaucoma surgery. Different mechanisms may cause the bleeding and there are several techniques available to approach this complication. Pre-Descemet hematoma should be drained to avoid permanent corneal stain and air/gas tamponade may help to prevent recurrences.

European real-world data about the use of a new delivery system containing a preservative-free multi-dose glaucoma treatment.

BACKGROUND: Glaucoma treatments are mostly presented in uni-dose or multi-dose format. A certain number of patients with visual acuity and dexterity problems may have problems in instilling eye drops. AIM: To assess patient satisfaction and ease of use of a preservative-free glaucoma treatment (dorzolamide/timolol) in a new and innovative patented multi-dose delivery system. METHODS: Retrospective, international, multicentre, non-interventional study in 788 adult patients using a multi-dose delivery system for at least 28 days. RESULTS: Mean patient age was 68.1 ± 12.1 years. Mean duration of multi-dose delivery system use was 132.1 ± 125.1 days; 66.5% of the patients previously used multi-dose and 33.5% uni-dose delivery systems (n = 734); 78.3% of the patients were satisfied or very satisfied with the multi-dose delivery system. A significant majority (all p ≤ 0.045) of patients with a QuickDash® score between [0 to 25[ (66.4%, n = 211) and [50 to 75[ (81.8%, n = 11) rated multi-dose delivery system as easy or very easy to open and significantly more subjects in the [0 to 25[ (72%) score group rated multi-dose delivery system as being better or much better than their previous device (n = 211). Significantly (all p < 0.01) more subjects with available visual acuity results rated multi-dose delivery system as good, better or much better than their previous dispensing device. CONCLUSION: The tested multi-dose delivery system was highly accepted. It is, therefore, suitable for glaucoma patients with decreased visual acuity and/or dexterity problems. Further studies may be necessary to assess the easiness of use of this easy-to-grip delivery system.

Phenotypic Description of the Spanish Multicentre Genetic Glaucoma Group Cohort.

INTRODUCTION: The aim of the study was to make a phenotypic description of the Spanish multicentre glaucoma group cohort of patients. DESIGN: Retrospective, observational, multicentre, cohort study. MATERIAL AND METHODS: The clinical charts of 152 patients with hereditary glaucoma from18 Spanish eye centres were reviewed in order to make an epidemiologic description of the type of glaucoma and associated factors. True hereditary cases were compared with familiar cases according to the Gong et al. criteria. RESULTS: 61% were true hereditary cases and 39% familiar cases. Ocular comorbidity, optic disc damage, and visual field mean defect were significantly more severe in hereditary patients, who required significantly more first-line hypotensive drugs and surgical interventions to control intraocular pressure than familiar patients. CONCLUSIONS: The strength of the hereditary component of glaucoma seems to worsen the clinical course, causing more structural and functional damage and requiring more intense glaucoma treatment. The family history of glaucoma should be carefully investigated and taken into consideration when making treatment decisions or intensifying previous treatment.

Effect of nepafenac on the foveal profile of glaucomatous patients undergoing phacoemulsification.

PURPOSE: Retrospective, pilot study to determine whether nepafenac treatment pre- and postcataract surgery in glaucoma patients using topical hypotensive agents minimized cystoid macular edema by comparing pre- and postsurgical foveal characteristics, as in some cases these agents cannot be withdrawn and, hypothetically, their inflammatory effect on the fovea could be neutralized by the addition of nepafenac. METHODS: Patients were divided into two subgroups depending on whether or not topical nepafenac was added to the surgical protocol (NEP = nepafenac group and nNEP = non nepafenac group). All had undergone phacoemulsification and data on pre- and postoperative macular status were recorded. RESULTS: In the nNEP group, there was a significant increase in foveal thickness (FT) in the first month postoperative visit with respect to the preoperative status (p = 0.006), and this situation did not change at the third postoperative month (p = 0.9411). In the NEP group, the increase in FT was not significant at the first month after surgery (p = 0.056) nor at the final visit (p = 0.268), in contrast to the nNEP group. CONCLUSION: This study of the possible prophylactic effect of nepafenac on postoperative macular edema supports the results of other studies that confirm subclinical edema post phacoemulsification, and found a significantly lower gradient in the increase in FT in patients treated pre- and postoperatively with nepafenac.

Persistent Hypotony Associated with Immunosuppressive Therapy in Glaucoma Drainage Implant Surgery.

PURPOSE: To describe the histopathology of non-valved implant capsules in three cases of persistent postoperative hypotony after the restrictive tube ligature was released in patients receiving immunosuppressive therapy. OBSERVATIONS: The macroscopic appearance of the capsules 3 and 4 months postoperatively was immature and loose. Microscopic examination disclosed extremely irregular thin tissue, with thicknesses ranging from 0.02 to 0.6 mm, depending on the capsular location studied. Withdrawal of immunosuppressive therapy did not facilitate rebuilding of new capsules. Replacement with a valved implant device was necessary in two cases; the third case recovered with tapering of prednisone. CONCLUSIONS AND IMPORTANCE: The use of chronic systemic immunosuppressive therapy might interfere with capsular formation around the plates of drainage devices inducing persistent hypotony. In these cases, the use of valved implants might be safer.

Survey of familial glaucoma shows a high incidence of cytochrome P450, family 1, subfamily B, polypeptide 1 (CYP1B1) mutations in non-consanguineous congenital forms in a Spanish population.

PURPOSE: To identify myocilin (MYOC) and cytochrome P450, family 1, subfamily B, polypeptide 1 (CYP1B1) mutations in a Spanish population with different clinical forms of familial glaucoma or ocular hypertension (OHT). METHODS: Index patients from 226 families participated in this study. Patients were diagnosed with familial glaucoma or OHT by complete ophthalmologic examination. Screening for MYOC mutations was performed in 207 index patients: 96 with adult-onset primary open-angle glaucoma (POAG), 21 with primary congenital glaucoma (PCG), 18 with juvenile-onset open-angle glaucoma (JOAG), five with Axenfeld-Rieger syndrome (ARS), and 67 with other types of glaucoma. One hundred two of the families (including all those in whom a MYOC mutation was detected) were also screened for CYP1B1 mutations: 45 POAG, 25 PCG, 21 JOAG, four ARS, and seven others. RESULTS: We examined 292 individuals (patients and relatives) with a positive family history of glaucoma or OHT. We identified two novel MYOC variants, p.Lys39Arg and p.Glu218Lys, in two families with POAG, and six previously reported MYOC mutations in seven families with POAG (four), JOAG (one), PCG (one), and normotensive glaucoma (one). CYP1B1 mutations were found in 16 index patients with PCG (nine), POAG (three), JOAG (two), and ARS (two). CONCLUSIONS: The high percentage (9/25=36%) of mutations in CYP1B1 found in non-consanguineous patients with congenital glaucoma mandates genetic testing. However, the percentage of mutations (9/207=4.4%) in MYOC associated with glaucoma is relatively low in our population. The variable phenotype expression of glaucoma, even in families, cannot be explained with a digenic mechanism between MYOC and CYP1B1.

Spanish multicenter tafluprost tolerability study.

AIM: To evaluate the ocular surface symptoms and signs associated with preservative-free 0.0015% tafluprost in patients with glaucoma or ocular hypertension (OHT). METHODS: Prospective non-interventional, multicentre, observational study on 134 patients, naive or on previous treatment with another prostaglandin analogue. In each visit (V1 baseline visit, V2 at 1 month and V3 at 3 months), patients evaluated five ocular surface symptoms as: absent, mild, moderate and severe. Parallelly, the ophthalmologist assessed the tear break-up time, keratitis, conjunctival hyperaemia, blepharitis, Schirmer test and tear meniscus. RESULTS: Patients with OHT (n=71, 53%) experienced a statistically significant improvement of all symptoms: stinging/burning/irritation, itching, foreign body sensation, tearing and dryness sensation at V3, while glaucoma patients improved all symptoms at both V2 and V3. In patients with OHT, all signs except Schirmer test improved and the decrease in hyperaemia was statistically significant. Eyes with glaucoma ameliorated the keratitis, hyperaemia and tear meniscus at V2 and V3 and the break-up time and blepharitis at V3. In the subset of patients with previous treatment (n=79, 58.9%), patients with OHT presented significant improvement of hyperaemia, yet the rest of signs did not decrease significantly or remained unchanged, while in patients with glaucoma all signs improved significantly at both visits. The intraocular pressure (IOP) drop in naive eyes was 22.2% (24.7-19.7 mm Hg) in OHT and 29.5% (33.7-25.3 mm Hg) in glaucoma eyes. In previously treated eyes, no statistically significant change in IOP was found. CONCLUSION: Preservative-free tafluprost is a well tolerated hypotensive agent that can be used in eyes with surface problems and in naive eyes.

One-year analysis of the iStent trabecular microbypass in secondary glaucoma.

PURPOSE: To evaluate the midterm efficacy and safety of the iStent(®) glaucoma device in patients with secondary open-angle glaucoma. PATIENTS AND METHODS: A prospective, nonrandomized, interventional case series involving 10 patients with secondary open-angle glaucoma (traumatic, steroid, pseudoexfoliative, and pigmentary glaucoma) of recent onset who underwent ab interno implantation iStent. Patients were assessed following the procedure on days 1, 7, and 15 and months 1, 3, 6, and 12, and examinations included visual acuity, intraocular pressure (IOP) measurement using Goldmann tonometry, number of glaucoma medications, and complications. Wilcoxon rank-test for data with abnormal distribution was used for the analysis of IOP and glaucoma medications at baseline versus 3, 6, and 12 months following the procedure. RESULTS: The mean baseline IOP was 26.5 ± 7.9 (range 18-40) mmHg, and significantly decreased in 10.4 ± 9.2 mmHg at three months (P < 0.05), in 7.4 ± 4.9 mmHg at six months (P < 0.05), and in 6.6 ± 5.4 mmHg at 12 months (P < 0.05) following iStent implantation. The mean number of hypotensive medications at baseline was 2.9 ± 0.7 (range 2-4). Statistically significant reductions in the number of medications of 1.1 ± 1.1 were observed at three months (P < 0.05), 1.0 ± 0.7 at six months (P < 0.05), and 1.1 ± 0.6 at 12 months (P < 0.05). No significant changes in visual acuity were noted. The most common complications comprised mild hyphema in seven eyes and transient IOP ≥30 mmHg in three eyes on postoperative day 1. Obstruction of the lumen of the stent with a blood clot was seen in three eyes, and all instances resolved spontaneously. CONCLUSION: The iStent is a safe and effective treatment option in patients with secondary open-angle glaucoma, and reduces the topical treatment burden in one hypotensive medication.

Intracameral bevacizumab (Avastin) for neovascular glaucoma: a pilot study in 6 patients.

PURPOSE: To describe the use of intracameral bevacizumab (ICB) Avastin in neovascular glaucoma (NVG) as the first maneuver before pan retinal photocoagulation and/or filtering surgery. METHODS: Between June 2006 and May 2007, 6 consecutive patients with NVG underwent intracameral injection of bevacizumab (1.25 mg/0.05 mL) as the initial treatment of NVG. Pre-ICB and post-ICB anterior segment photography, iris fluoresceingraphy when possible, gonioscopy with peripheral anterior synechiae (PAS), neovascular membrane (NVM) extension grading, as well as intraocular pressure (IOP) changes during treatment were recorded. All patients were followed for at least 7 months. RESULTS: ICB resulted in a marked regression of anterior segment neovascularization with IOP control without filtering surgery in 2 cases. When PAS extended <330 degrees without previous glaucoma, no filtering surgery was needed to control IOP<18 mm Hg. Iris neovascularization extension had no prognostic value in terms of IOP control. After vascular regression following the administration of ICB, filtering surgery with drainage implants or trabeculectomy were performed when needed with no added difficulties owing to the underlying NVG. No macroscopic signs of corneal toxicity were detected, even when ICB injection had to be repeated. In this case, the time elapsed for the neovascular membrane to reappear at the anterior segment was 3 months. CONCLUSION: ICB resulted in a rapid regression of the iris and angle neovascularization, which permitted to halt the progression of PAS process. This pilot study shows that intracameral injection of bevacizumab may be a helpful adjunct for the surgical treatment of NVG.