Tammy L. Yanovitch

Mutations in SCO2 Are Associated with Autosomal-Dominant High-Grade Myopia

Myopia, or near-sightedness, is an ocular refractive error of unfocused image quality in front of the retinal plane. Individuals with high-grade myopia (dioptric power greater than -6.00) are predisposed to ocular morbidities such as glaucoma, retinal detachment, and myopic maculopathy. Nonsyndromic, high-grade myopia is highly heritable, and to date multiple gene loci have been reported. We performed exome sequencing in 4 individuals from an 11-member family of European descent from the United States. Affected individuals had a mean dioptric spherical equivalent of -22.00 sphere. A premature stop codon mutation c.157C>T (p.Gln53*) cosegregating with disease was discovered within SCO2 that maps to chromosome 22q13.33. Subsequent analyses identified three additional mutations in three highly myopic unrelated individuals (c.341G>A, c.418G>A, and c.776C>T). To determine differential gene expression in a developmental mouse model, we induced myopia by applying a -15.00D lens over one eye. Messenger RNA levels of SCO2 were significantly downregulated in myopic mouse retinae. Immunohistochemistry in mouse eyes confirmed SCO2 protein localization in retina, retinal pigment epithelium, and sclera. SCO2 encodes for a copper homeostasis protein influential in mitochondrial cytochrome c oxidase activity. Copper deficiencies have been linked with photoreceptor loss and myopia with increased scleral wall elasticity. Retinal thinning has been reported with an SC02 variant. Human mutation identification with support from an induced myopic animal provides biological insights of myopic development.

Angiopoietin receptor TEK mutations underlie primary congenital glaucoma with variable expressivity

Primary congenital glaucoma (PCG) is a devastating eye disease and an important cause of childhood blindness worldwide. In PCG, defects in the anterior chamber aqueous humor outflow structures of the eye result in elevated intraocular pressure (IOP); however, the genes and molecular mechanisms involved in the etiology of these defects have not been fully characterized. Previously, we observed PCG-like phenotypes in transgenic mice that lack functional angiopoietin-TEK signaling. Herein, we identified rare TEK variants in 10 of 189 unrelated PCG families and demonstrated that each mutation results in haploinsufficiency due to protein loss of function. Multiple cellular mechanisms were responsible for the loss of protein function resulting from individual TEK variants, including an absence of normal protein production, protein aggregate formation, enhanced proteasomal degradation, altered subcellular localization, and reduced responsiveness to ligand stimulation. Further, in mice, hemizygosity for Tek led to the formation of severely hypomorphic Schlemms canal and trabecular meshwork, as well as elevated IOP, demonstrating that anterior chamber vascular development is sensitive to Tek gene dosage and the resulting decrease in angiopoietin-TEK signaling. Collectively, these results identify TEK mutations in patients with PCG that likely underlie disease and are transmitted in an autosomal dominant pattern with variable expressivity.

CYP1B1, MYOC, and LTBP2 Mutations in Primary Congenital Glaucoma Patients in the United States

PURPOSE To screen primary congenital glaucoma patients in the United States for sequence variants within the CYP1B1, LTBP2, and MYOC genes using Sanger and whole exome sequencing. DESIGN Retrospective case-control study. METHODS Fifty-seven primary congenital glaucoma patients (47 families), 71 unaffected family members of the primary congenital glaucoma probands, and 101 healthy unrelated individuals were recruited from a single institution. Sanger sequencing of the primary congenital glaucoma gene, CYP1B1, was performed on 47 proband deoxyribonucleic acid samples. Simultaneously, whole exome sequencing was conducted on 3 families, each including more than 1 affected individual. Concurrently, 33 of 47 primary congenital glaucoma probands with extended family deoxyribonucleic acid samples were screened for LTBP2 and MYOC gene mutations. Exome-sequenced variations were validated by additional Sanger sequencing to confirm segregation of filtered disease-causing single nucleotide variations. RESULTS Seven primary congenital glaucoma families (14.9%) manifested disease phenotypes attributable to CYP1B1 mutations. One primary congenital glaucoma family possessed homozygous mutant alleles, whereas 6 families carried compound heterozygous mutations. Five novel combinations of compound heterozygous mutations were identified, of which 2 combinations were found with whole exome sequencing. No disease-causing mutations within the LTBP2 and MYOC genes were discovered. CONCLUSIONS This study analyzed CYP1B1, LTBP2, and MYOC mutations in a cohort of primary congenital glaucoma patients from the United States, applying whole exome sequencing as a complementary tool to Sanger sequencing. Whole exome sequencing, coupled with Sanger sequencing, may identify novel genes in primary congenital glaucoma patients who have no mutations in known primary congenital glaucoma genes.

Clinical and histologic ocular findings in pompe disease.

PURPOSE Limited information is available on the ocular findings in patients with Pompe disease. METHOD This study summarizes this information with a systematic literature review; reports the ocular histologic findings seen in a deceased infant with Pompe disease who was receiving enzyme replacement therapy and in a deceased adult with late-onset Pompe disease; and notes the new observation of ptosis in children with infantile-onset Pompe disease who are receiving enzyme replacement therapy. RESULTS Six articles were found on the ultrastructural-histopathologic eye findings in Pompe disease. Previously reported clinical ocular findings included strabismus and ptosis. Glycogen accumulation and vacuolar myopathy have been seen histologically. CONCLUSION Based on these clinical and histologic reports, patients with Pompe disease may have an increased incidence of ocular abnormalities, such as ptosis and strabismus, and therefore should undergo ophthalmologic examination.

Latanoprost in pediatric glaucoma—pediatric exposure over a decade

BACKGROUND Although numerous studies of latanoprost in adult glaucoma have shown it to be an effective hypotensive agent with a low incidence of side effects, these issues have not been well studied in pediatric glaucomas. The purpose of the current study is to evaluate the safety and intraocular pressure (IOP) lowering effect of latanoprost in various pediatric glaucomas over a long period. SUBJECTS AND METHODS This retrospective study included all children treated with latanoprost at our institution from 1996 to 2007. Demographic, glaucoma-related, and side-effect information was recorded for each subject. Duration of latanoprost exposure was calculated in child-months (1 child exposed for 1 month). If interpretable IOP data were available, the presence or absence of a treatment response (IOP reduction > or =15% from baseline) was determined for each subject. RESULTS A total of 115 subjects with latanoprost exposure were identified, with a collective exposure of 2,325 child-months. Exposure for > or =1 year occurred in 52 subjects. Side effects were mild and infrequently reported. Of the 115 subjects, 63 had interpretable IOP data, and 22 (35%) were treatment responders. Predictors of a response included a diagnosis of juvenile open-angle glaucoma, monotherapy, and older age. CONCLUSIONS This large study of latanoprost-treated children confirms the excellent safety profile of the drug in the treatment of pediatric glaucoma. The study also confirms latanoprosts IOP-lowering ability in older children with juvenile open-angle glaucoma and in some children with aphakic glaucoma. Prospective studies are needed to better define the optimal role of latanoprost in the treatment of pediatric glaucoma, especially congenital glaucoma.

Genetic Variants Associated With Severe Retinopathy of Prematurity in Extremely Low Birth Weight Infants

PURPOSE To determine genetic variants associated with severe retinopathy of prematurity (ROP) in a candidate gene cohort study of US preterm infants. METHODS Preterm infants in the discovery cohort were enrolled through the Eunice Kennedy Shriver National Institute of Child Health and Human Development Neonatal Research Network, and those in the replication cohort were from the University of Iowa. All infants were phenotyped for ROP severity. Because of differences in the durations of enrollment between cohorts, severe ROP was defined as threshold disease in the discovery cohort and as threshold disease or type 1 ROP in the replication cohort. Whole genome amplified DNA from stored blood spot samples from the Neonatal Research Network biorepository was genotyped using an Illumina GoldenGate platform for candidate gene single nucleotide polymorphisms (SNPs) involving angiogenic, developmental, inflammatory, and oxidative pathways. Three analyses were performed to determine significant epidemiologic variables and SNPs associated with levels of ROP severity. Analyses controlled for multiple comparisons, ancestral eigenvalues, family relatedness, and significant epidemiologic variables. Single nucleotide polymorphisms significantly associated with ROP severity from the discovery cohort were analyzed in the replication cohort and in meta-analysis. RESULTS Eight hundred seventeen infants in the discovery cohort and 543 in the replication cohort were analyzed. Severe ROP occurred in 126 infants in the discovery and in 14 in the replication cohort. In both cohorts, ventilation days and seizure occurrence were associated with severe ROP. After controlling for significant factors and multiple comparisons, two intronic SNPs in the gene BDNF (rs7934165 and rs2049046, P < 3.1 × 10(-5)) were associated with severe ROP in the discovery cohort and were not associated with severe ROP in the replication cohort. However, when the cohorts were analyzed together in an exploratory meta-analysis, rs7934165 increased in associated significance with severe ROP (P = 2.9 × 10(-7)). CONCLUSIONS Variants in BDNF encoding brain-derived neurotrophic factor were associated with severe ROP in a large candidate gene study of infants with threshold ROP.

The accuracy of photoscreening at detecting treatable ocular conditions in children with Down syndrome

BACKGROUND Children with Down syndrome (DS) have an increased prevalence of ocular disorders, including amblyopia, strabismus, and refractive error. Health maintenance guidelines from the Down Syndrome Medical Interest Group recommend ophthalmologic examinations every 1 to 2 years for these children. Photoscreening may be a cost-effective option for subsequent screening evaluations after an initial complete examination, but no study has evaluated the accuracy of photoscreening in children with DS. The purpose of this study is to determine the sensitivity, specificity, and positive and negative predictive values of photoscreening in detecting treatable ocular conditions in children with DS. METHODS Photoscreening and complete ophthalmologic evaluations were performed in 50 consecutive 3- to 10-year-old children with DS. Sensitivity, specificity, and positive and negative predictive values were calculated with the use of ophthalmologic examination findings as the reference standard. RESULTS Most children were able to complete photoscreening (94% with Medical Technology and Innovations [MTI] and 90% with Visiscreen OSS-C [VR]). Many children had an identified diagnosis on ophthalmologic examination (n = 46, 92%). Of these, approximately one-half (n = 27, 54%) had one or more condition(s) requiring treatment. Both the MTI and VR photoscreening devices had a sensitivity of 93% (95% confidence interval 0.76-0.99) for detecting treatable ocular conditions. The specificities for the MTI and VR photoscreening were 0.35 (0.18-0.57) and 0.55 (0.34-0.74), respectively. CONCLUSIONS Photoscreening is sensitive but less specific at detecting treatable ocular conditions in children with DS. In specific instances, the use of photoscreening in the DS population has the potential to save time and expense related to routine eye examinations, particularly in children with a normal baseline comprehensive examination.

Tortuosity of arterioles and venules in quantifying plus disease

BACKGROUND Plus disease is the major criterion for laser treatment of retinopathy of prematurity. ROPtool is a computer program that traces retinal blood vessels and measures their tortuosity. Our objectives were to determine (1) whether examiners could accurately discriminate between arterioles and venules and (2) whether tortuosity sufficient for plus disease and pre-plus disease was assessed most accurately by considering arterioles, venules, or both. METHODS One hundred retinal vessels were identified in 25 images randomly selected from 184 total images. Three pediatric ophthalmologists independently designated vessels as arteriole or venule. Seventy-seven images that had at least 1 traceable arteriole and venule in each quadrant were analyzed by ROPtool, and the results were compared with the consensus of 3 expert examiners. Receiver operating characteristics (ROC) curves were generated and areas under the curves calculated to quantify the diagnostic utility of ROPtools assessment of tortuosity of arterioles, venules, and both. RESULTS Three pediatric ophthalmologists agreed on the designation of arteriole or venule for 83 of 100 blood vessels. With the use of expert consensus as the reference standard, areas under the ROC curves for identification of tortuosity sufficient for plus disease were 0.91, 0.70, and 0.93 for arterioles, venules, and both, respectively. Areas under the ROC curves for identification of tortuosity sufficient for pre-plus disease were 0.91, 0.63, and 0.90 for arterioles, venules, and both, respectively. CONCLUSIONS When considering whether tortuosity is sufficient for plus or pre-plus disease, the assessment of either arterioles alone or of arterioles and venules together resulted in high diagnostic accuracy.

Angiopoietin-1 is required for Schlemm’s canal development in mice and humans

Primary congenital glaucoma (PCG) is a leading cause of blindness in children worldwide and is caused by developmental defects in 2 aqueous humor outflow structures, Schlemm’s canal (SC) and the trabecular meshwork. We previously identified loss-of-function mutations in the angiopoietin (ANGPT) receptor TEK in families with PCG and showed that ANGPT/TEK signaling is essential for SC development. Here, we describe roles for the major ANGPT ligands in the development of the aqueous outflow pathway. We determined that ANGPT1 is essential for SC development, and that Angpt1-knockout mice form a severely hypomorphic canal with elevated intraocular pressure. By contrast, ANGPT2 was dispensable, although mice deficient in both Angpt1 and Angpt2 completely lacked SC, indicating that ANGPT2 compensates for the loss of ANGPT1. In addition, we identified 3 human subjects with rare ANGPT1 variants within an international cohort of 284 PCG patients. Loss of function in 2 of the 3 patient alleles was observed by functional analysis of ANGPT1 variants in a combined in silico, in vitro, and in vivo approach, supporting a causative role for ANGPT1 in disease. By linking ANGPT1 with PCG, these results highlight the importance of ANGPT/TEK signaling in glaucoma pathogenesis and identify a candidate target for therapeutic development.

Travoprost in children: adverse effects and intraocular pressure response.

Because of the limited ability to perform controlled, randomized studies in children, the safety and effectiveness of topical medications in pediatric glaucoma is sometimes difficult to determine. Although travoprost has been commercially available since 2001, there are no published reports on its use in children. This retrospective study found travoprost to have minimal adverse events in children and to reduce IOP in select cases of pediatric glaucoma.