Whole-Exome Sequencing of 21 Families: Candidate Genes for Early-Onset High Myopia.

High myopia is the most severe and pathological form of myopia. It occurs when the spherical refractive error exceeds -6.00 spherical diopters (SDs) or the axial length (AL) of the eye is greater than 26 mm. This article focuses on early-onset high myopia, an increasingly common condition that affects children under 10 years of age and can lead to other serious ocular pathologies. Through the genetic analysis of 21 families with early-onset high myopia, this study seeks to contribute to a better understanding of the role of genetics in this disease and to propose candidate genes. Whole-exome sequencing studies with a panel of genes known to be involved in the pathology were performed in families with inconclusive results: 3% of the variants found were classified as pathogenic, 6% were likely pathogenic and the remaining 91% were variants of uncertain significance. Most of the families in this study were found to have alterations in several of the proposed genes. This suggests a polygenic inheritance of the pathology due to the cumulative effect of the alterations. Further studies are needed to validate and confirm the role of these alterations in the development of early-onset high myopia and its polygenic inheritance.

Whole-Exome Sequencing of 24 Spanish Families: Candidate Genes for Non-Syndromic Pediatric Keratoconus.

Keratoconus is a corneal dystrophy that is one of the main causes of corneal transplantation and for which there is currently no effective treatment for all patients. The presentation of this disease in pediatric age is associated with rapid progression, a worse prognosis and, in 15-20% of cases, the need for corneal transplantation. It is a multifactorial disease with genetic variability, which makes its genetic study difficult. Discovering new therapeutic targets is necessary to improve the quality of life of patients. In this manuscript, we present the results of whole-exome sequencing (WES) of 24 pediatric families diagnosed at the University Hospital La Paz (HULP) in Madrid. The results show an oligogenic inheritance of the disease. Genes involved in the structure, function, cell adhesion, development and repair pathways of the cornea are proposed as candidate genes for the disease. Further studies are needed to confirm the involvement of the candidate genes described in this article in the development of pediatric keratoconus.

Whole Exome Sequencing of 20 Spanish Families: Candidate Genes for Non-Syndromic Pediatric Cataracts.

Non-syndromic pediatric cataracts are defined as opacification of the crystalline lens that occurs during the first years of life without affecting other organs. Given that this disease is one of the most frequent causes of reversible blindness in childhood, the main objective of this study was to propose new responsible gene candidates that would allow a more targeted genetic approach and expand our genetic knowledge about the disease. We present a whole exome sequencing (WES) study of 20 Spanish families with non-syndromic pediatric cataracts and a previous negative result on an ophthalmology next-generation sequencing panel. After ophthalmological evaluation and collection of peripheral blood samples from these families, WES was performed. We were able to reach a genetic diagnosis in 10% of the families analyzed and found genes that could cause pediatric cataracts in 35% of the cohort. Of the variants found, 18.2% were classified as pathogenic, 9% as likely pathogenic, and 72.8% as variants of uncertain significance. However, we did not find conclusive results in 55% of the families studied, which suggests further studies are needed. The results of this WES study allow us to propose LONP1, ACACA, TRPM1, CLIC5, HSPE1, ODF1, PIKFYVE, and CHMP4A as potential candidates to further investigate for their role in pediatric cataracts, and AQP5 and locus 2q37 as causal genes.

OPA1 Dominant Optic Atrophy: Diagnostic Approach in the Pediatric Population.

A clinical and genetic study was conducted with pediatric patients and their relatives with optic atrophy 1 (OPA1) mutations to establish whether there is a genotype-phenotype correlation among the variants detected within and between families. Eleven children with a confirmed OPA1 mutation were identified during the study period. The main initial complaint was reduced visual acuity (VA), present in eight patients of the cohort. Eight of eleven patients had a positive family history of optic atrophy. The mean visual acuity at the start of the study was 0.40 and 0.44 LogMAR in the right and left eye, respectively. At the end of the study, the mean visual acuity was unchanged. Optical coherence tomography during the first visit showed a mean retinal nerve fiber layer thickness of 81.6 microns and 80.5 microns in the right and left eye, respectively; a mean ganglion cell layer of 52.5 and 52.4 microns, respectively, and a mean central macular thickness of 229.5 and 233.5 microns, respectively. The most common visual field defect was a centrocecal scotoma, and nine out of eleven patients showed bilateral temporal disc pallor at baseline. Sequencing of OPA1 showed seven different mutations in the eleven patients, one of which, NM_130837.3: c.1406_1407del (p.Thr469LysfsTer16), has not been previously reported. Early diagnosis of dominant optic atrophy is crucial, both for avoiding unnecessary consultations and/or treatments and for appropriate genetic counseling.

Next-Generation Sequencing Screening of 43 Families with Non-Syndromic Early-Onset High Myopia: A Clinical and Genetic Study.

Early-onset high myopia (EoHM) is a disease that causes a spherical refraction error of ≥-6 diopters before 10 years of age, with potential multiple ocular complications. In this article, we report a clinical and genetic study of 43 families with EoHM recruited in our center. A complete ophthalmological evaluation was performed, and a sample of peripheral blood was obtained from proband and family members. DNA was analyzed using a customized next-generation sequencing panel that included 419 genes related to ophthalmological disorders with a suspected genetic cause, and genes related to EoHM pathogenesis. We detected pathogenic and likely pathogenic variants in 23.9% of the families and detected variants of unknown significance in 76.1%. Of these, 5.7% were found in genes related to non-syndromic EoHM, 48.6% in genes associated with inherited retinal dystrophies that can include a syndromic phenotype, and 45.7% in genes that are not directly related to EoHM or retinal dystrophy. We found no candidate genes in 23% of the patients, which suggests that further studies are needed. We propose a systematic genetic analysis for patients with EoHM because it helps with follow-up, prognosis and genetic counseling.

Neovascular Glaucoma in Children: A case series and a review of the literature.

PURPOSE: To study the uncommon causes and treatment options for neovascular glaucoma in children. PATIENTS AND METHODS: A review of the literature on neovascular glaucoma in children was conducted and we present three cases of neovascular glaucoma in children. RESULTS: We present three cases of neovascular glaucoma: two cases were secondary to a retinal vasoproliferative tumor-one to neurofibromatosis type 1 and the other to exudative retinopathy secondary to mild retinopathy of prematurity-and one case was secondary to a central retina vein occlusion secondary to an optic nerve glioma. Vision in the affected eye was severely impaired in all the children. CONCLUSION: The diagnosis and treatment of neovascular glaucoma in children is challenging and often a complication of a systemic or late-stage ocular condition. An appropriate diagnosis and estimation of the visual potential are essential to determine the correct treatment, especially in young children.

The frequency of severe corneal opacities in premature infants.

PURPOSE: To study the frequency and possible causes of severe corneal opacities in premature infants undergoing retinopathy of prematurity (ROP) screening in the neonatal intensive care unit (NICU) at a single institution. METHODS: The medical records of all infants screened for ROP in the NICU between January 2015 and March 2019 were reviewed retrospectively. Criteria for screening were gestational age (GA) of <32 weeks or birth weight (BW) <1501 g. Characteristics of premature infants with severe corneal opacities were extracted from the record. RESULTS: A total of 445 premature infants were screened during the 51-month period. The prevalence of severe corneal opacities was 1.4% (6 infants). The median GA of the affected infants was 24.5 weeks (range, 23-32), and the mean BW was 624 g (range, 500-860 g). Two infants had lagophthalmos; their corneal opacity was unilateral. All the infants were under continuous positive airway pressure (CPAP) for a prolonged period. All 4 infants with bilateral corneal opacities required treatment for severe ROP-laser (n =1), bevacizumab injection (n = 1), or both (n = 2). CONCLUSIONS: Severe corneal opacities in premature infants are rare but potentially sight threatening, because they can hinder the retinal examination. Lagophthalmos-related exposure keratopathy is an obvious risk factor. CPAP therapy may be another risk factor leading to the development of severe corneal opacities. Identification of infants at risk and prompt commencement of lubricants is necessary to avoid long-term corneal opacities.

Expanding the Phenotypic Spectrum of PAX6 Mutations: From Congenital Cataracts to Nystagmus.

BACKGROUND: Congenital aniridia is a complex ocular disorder, usually associated with severe visual impairment, generally caused by mutations on the PAX6 gene. The clinical phenotype of PAX6 mutations is highly variable, making the genotype-phenotype correlations difficult to establish. METHODS: we describe the phenotype of eight patients from seven unrelated families with confirmed mutations in PAX6, and very different clinical manifestations. RESULTS: Only two patients had the classical aniridia phenotype while the other two presented with aniridia-related manifestations, such as aniridia-related keratopathy or partial aniridia. Congenital cataracts were the main manifestation in three of the patients in this series. All the patients had nystagmus and low visual acuity. CONCLUSIONS: The diagnosis of mild forms of aniridia is challenging, but these patients have a potentially blinding hereditary disease that might present with a more severe phenotype in future generations. Clinicians should be aware of the mild aniridia phenotype and request genetic testing to perform an accurate diagnosis.

Molecular and Genetic Mechanism of Non-Syndromic Congenital Cataracts. Mutation Screening in Spanish Families.

Our purpose was to identify mutations responsible for non-syndromic congenital cataracts through the implementation of next-generation sequencing (NGS) in our center. A sample of peripheral blood was obtained from probands and willing family members and genomic DNA was extracted from leukocytes. DNA was analyzed implementing a panel (OFTv2.1) including 39 known congenital cataracts disease genes. 62 probands from 51 families were recruited. Pathogenic or likely pathogenic variants were identified in 32 patients and 25 families; in 16 families (64%) these were de novo mutations. The mutation detection rate was 49%. Almost all reported mutations were autosomal dominant. Mutations in crystallin genes were found in 30% of the probands. Mutations in membrane proteins were detected in seven families (two in GJA3 and five in GJA8). Mutations in LIM2 and MIP were each found in three families. Other mutations detected affected EPHA2, PAX6, HSF4 and PITX3. Variants classified as of unknown significance were found in 5 families (9.8%), affecting CRYBB3, LIM2, EPHA2, ABCB6 and TDRD7. Mutations lead to different cataract phenotypes within the same family.

Primary Immunodeficiency Diseases Presenting with Chalazia as the First Manifestation.

Purpose: Ocular manifestations in primary immunodeficiency diseases are rare, but they can be the initial manifestation. This can lead to the prompt diagnosis and treatment of the disease and achieve a reduction of severe systemic complications.Case Report: We present two cases where a recurrent giant chalazion was the symptom that led to the diagnosis and early treatment of a patient with X-linked chronic granulomatous disease (CGD), and a patient with hyperimmunoglobulin E syndrome.Conclusion: Even though chalazia are common and benign, children presenting with recurrent giant chalazia or torpid evolution after surgery should be investigated for immunodeficiencies to reduce the severe and potentially fatal complications of the disease.

New Normative Database of Inner Macular Layer Thickness Measured by Spectralis OCT Used as Reference Standard for Glaucoma Detection.

PURPOSE: This study examines the capacity to detect glaucoma of inner macular layer thickness measured by spectral-domain optical coherence tomography (SD-OCT) using a new normative database as the reference standard. METHODS: Participants (N = 148) were recruited from Leuven (Belgium) and Zaragoza (Spain): 74 patients with early/moderate glaucoma and 74 age-matched healthy controls. One eye was randomly selected for a macular scan using the Spectralis SD-OCT. The variables measured with the instrument's segmentation software were: macular nerve fiber layer (mRNFL), ganglion cell layer (GCL), and inner plexiform layer (IPL) volume and thickness along with circumpapillary RNFL thickness (cpRNFL). The new normative database of macular variables was used to define the cutoff of normality as the fifth percentile by age group. Sensitivity, specificity, and area under the receiver operating characteristic curve (AUROC) of each macular measurement and of cpRNFL were used to distinguish between patients and controls. RESULTS: Overall sensitivity and specificity to detect early-moderate glaucoma were 42.2% and 88.9% for mRNFL, 42.4% and 95.6% for GCL, 42.2% and 94.5% for IPL, and 53% and 94.6% for RNFL, respectively. The best macular variable to discriminate between the two groups of subjects was outer temporal GCL thickness as indicated by an AUROC of 0.903. This variable performed similarly to mean cpRNFL thickness (AUROC = 0.845; P = 0.29). CONCLUSIONS: Using our normative database as reference, the diagnostic power of inner macular layer thickness proved comparable to that of peripapillary RNFL thickness. TRANSLATIONAL RELEVANCE: Spectralis SD-OCT, cpRNFL thickness, and individual macular inner layer thicknesses show comparable diagnostic capacity for glaucoma and RNFL, GCL, and IPL thickness may be useful as an alternative diagnostic test when the measure of cpRNFL shows artifacts.

Impacts of age and sex on retinal layer thicknesses measured by spectral domain optical coherence tomography with Spectralis.

OBJECTIVE: To examine differences in individual retinal layer thicknesses measured by spectral domain optical coherence tomography (SD-OCT) (Spectralis®) produced with age and according to sex. DESIGN: Cross-sectional, observational study. METHODS: The study was conducted in 297 eyes of 297 healthy subjects aged 18 to 87 years. In one randomly selected eye of each participant the volume and mean thicknesses of the different macular layers were measured by SD-OCT using the instrument's macular segmentation software. MAIN OUTCOME MEASURES: Volume and mean thickness of macular retinal nerve fiber layer (mRNFL), ganglion cell layer (GCL), inner plexiform layer (IPL), inner nuclear layer (INL), outer plexiform layer (OPL), outer nuclear layer (ONL), retinal pigmentary epithelium (RPE) and photoreceptor layer (PR). RESULTS: Retinal thickness was reduced by 0.24 µm for every one year of age. Age adjusted linear regression analysis revealed mean GCL, IPL, ONL and PR thickness reductions and a mean OPL thickness increase with age. Women had significantly lower mean GCL, IPL, INL, ONL and PR thicknesses and volumes and a significantly greater mRNFL volume than men. CONCLUSION: The thickness of most retinal layers varies both with age and according to sex. Longitudinal studies are needed to determine the rate of layer thinning produced with age.

Utility of Bruch membrane opening-based optic nerve head parameters in myopic subjects.

PURPOSE: To evaluate whether the new rim analysis software with spectral-domain optical coherence tomography (SD-OCT) shows advantages over the retinal nerve fiber layer (RNFL) thickness in patients with moderate myopia. METHODS: In this prospective cross-sectional study, we studied 65 healthy subjects, 37 with spherical refractive errors in the range of -3 to -6 D (moderate, G1) and 28 with less than -3 D (low/non-myopic, G0). All patients were examined with Heidelberg Spectralis SD-OCT, including Glaucoma Premium Module Edition (GPME) software. With GPME, we analyzed the neuroretinal rim (Bruch membrane opening-minimum rim width [BMO-MRW]) and RNFL. RESULTS: The average age of subjects was 30.2 ± 9.3 years for G0 and 29.9 ± 7.1 years for G1 (p = 0.903). Mean sphere was -0.5 ± 0.3 D (-1.25 to 0 D) G0 and -3.9 ± 0.3 D (-6.00 to -3 D) G1 (p<0.001). The RNFL thickness comparison between G0 and G1 showed a significantly lower thickness in G1 (p = 0.018). The BMO-MRW measurements were similar in both groups (p = 0.331). With the BMO-MRW examination, the number of sectors classified as pathologic per subject in G1 were significantly lower compared to RNFL analysis (p = 0.023). CONCLUSIONS: Ring analysis based on BMO-MRW measurements shows a lower rate of false-positives compared to RNFL thickness when studying healthy moderate myopic eyes and it would be advisable to take this into consideration when analyzing these patients.

Normative database for separate inner retinal layers thickness using spectral domain optical coherence tomography in Caucasian population.

PURPOSE: Develop the first normative database of the thickness of every inner retinal layer in the macular area in a healthy, Caucasian population between 18 to 87 years old, using Spectralis Optical Coherence Tomography (OCT). METHODS: On this transversal, observational study, 300 patients between 18 to 87 years old and without an ophthalmological condition were recruited. Macular OCT scans were performed on all patients (Spectralis OCT, Heidelberg Engineering). An axial length measurement, and keratometry were performed using an optical biometer. The volume and thickness of the different macular sectors of the inner retinal layers (retinal nerve fiber layer (RNFL), ganglion cells layer (CGL) and inner plexiform layer (IPL)) were analyzed with the Spectralis OCT segmentation software. An eye was randomly selected for each patient. RESULTS: 297 patients (179 females and 118 males) were included in the study. The mean age was 56.07 years (range: 40.50-72). The multivariate analysis showed a positive correlation between the RNFL thickness and the axial length (p < 0.001). The mean central retinal thickness was 278.2 µm (range: 266-291), the mean central RNFL thickness was 12.61 µm (range: 11-14), the mean central CGL thickness was 17.63 µm (range: 14-21) and the mean central IPL thickness was 22.02 µm (range: 20-25). The multivariate analysis showed a negative correlation between age and CGL thickness and inner IPL thickness (p< 0.001). CONCLUSION: This study provides a normative database of the volume of each of the inner retinal layers on a Caucasian population.