ABSTRACT
Purpose: Homozygous hypomorphic variants of the RP1 gene, including c.5797C>T, p.Arg1933Ter, have traditionally been considered non-pathogenic. This study aimed to elucidate the clinical manifestations of late-onset, slowly progressive cone/macular dystrophy in patients homozygous for p.Arg1933Ter in the RP1 gene. Methods: Five patients with biallelic p.Arg1933Ter in RP1 were retrospectively recruited, and their clinical profiles were analyzed. Copy number variation analysis and Alu insertion assessment of genes associated with inherited retinal diseases were conducted. The results of comprehensive ophthalmological examinations, multimodal imaging, and full-field electroretinogram tests were analyzed. Results: No specific sequencing errors or structural variations associated with the clinical phenotypes were identified. Alu element insertion in RP1 was not detected. The mean ± SD age at the first visit was 62.2 ± 9.8 years, with symptoms typically starting between 45 and 50 years of age. Two patients exhibited a mild form of cone/macular dystrophy, characterized by a relatively preserved fundus appearance and blurring of the ellipsoid zone on optical coherence tomography. Three patients had late-onset cone/macular dystrophy with significant atrophy. Conclusions: To our knowledge, this study is the first to report that a homozygous hypomorphic variant of RP1, previously considered non-pathogenic, leads to cone/macular dystrophy. Translational Relevance: The study introduces novel possibilities suggesting that the homozygous hypomorphic variant of RP1 may be linked to variant pathogenicity.
Subject(s)
Electroretinography , Eye Proteins , Tomography, Optical Coherence , Humans , Male , Female , Middle Aged , Retrospective Studies , Aged , Eye Proteins/genetics , Visual Acuity , DNA Copy Number Variations/genetics , Disease Progression , Cone Dystrophy/genetics , Cone Dystrophy/diagnostic imaging , Macular Degeneration/genetics , Macular Degeneration/pathology , Macular Degeneration/diagnostic imaging , Macular Degeneration/congenital , Pedigree , Homozygote , Phenotype , Mutation , Adult , Age of Onset , Microtubule-Associated ProteinsABSTRACT
INTRODUCTION: The aim of this study was to evaluate the progression of atrophy as determined by spectral-domain optical coherence tomography (SD-OCT) in patients with molecularly confirmed ABCA4-associated Stargardt disease type 1 (STGD1) over a 24-month period in a multicenter prospective cohort study. METHODS: SD-OCT images from 428 eyes of 236 patients were analyzed. Change of mean thickness (MT) and intact area were estimated after semiautomated segmentation for the following individual layers in the central subfield (CS), inner ring (IR), and outer ring (OR) of the ETDRS grid: retinal pigment epithelium (RPE), outer segments (OSs), inner segments (IS), outer nuclear layer (ONL) inner retina (IR), and total retina. RESULTS: Statistically significant decreases of all outer retinal layers (RPE, OS, IS, and ONL) could be observed over a 24-month period both in decline of mean retinal thickness and intact area (p < 0.0001, respectively), whereas the IR showed an increase of retinal thickness in the CS and IR and remained unchanged in the OR. CONCLUSIONS: Significant loss could be detected in outer retinal layers by SD-OCT over a 24-month period in patients with STGD1. Loss of thickness and/or intact area of such layers may serve as potential endpoints for clinical trials that aim to slow down the disease progression of STGD1.
Subject(s)
Disease Progression , Macular Degeneration , Retinal Pigment Epithelium , Stargardt Disease , Tomography, Optical Coherence , Visual Acuity , Humans , Tomography, Optical Coherence/methods , Stargardt Disease/diagnosis , Male , Prospective Studies , Female , Adult , Young Adult , Middle Aged , Macular Degeneration/diagnosis , Macular Degeneration/congenital , Retinal Pigment Epithelium/pathology , Retinal Pigment Epithelium/diagnostic imaging , Adolescent , Follow-Up Studies , Retina/diagnostic imaging , Retina/pathology , ChildABSTRACT
PURPOSE: To investigate the temporal sequence of changes in the photoreceptor cell mosaic in patients with Stargardt disease type 1, using adaptive optics scanning laser ophthalmoscopy. METHODS: Two brothers with genetically confirmed Stargardt disease type 1 underwent comprehensive eye exams, spectral-domain optical coherence tomography, fundus autofluorescence, and adaptive optics scanning laser ophthalmoscopy imaging 3 times over the course of 28 months. Confocal images of the cones and rods were obtained from the central fovea to 10° inferiorly. Photoreceptors were counted in sampling windows at 100- µ m intervals of 200 µ m × 200 µ m for cones and 50 µ m × 50 µ m for rods, using custom cell marking software with manual correction. Photoreceptor density and spacing were measured and compared across imaging sessions using one-way analysis of variance. RESULTS: Adaptive optics scanning laser ophthalmoscopy revealed the younger brother had a 30% decline in foveal cone density after 8 months, followed by complete loss of foveal cones at 28 months; the older brother had no detectable foveal cones at baseline. In the peripheral macula, cone and rod spacings were greater than normal in both patients. The ratio of the cone spacing to rod spacing was greater than normal across all eccentricities, with a greater divergence closer to the foveal center. CONCLUSION: Cone cell loss may be an early pathogenetic step in Stargardt disease. Adaptive optics scanning laser ophthalmoscopy provides the capability to track individual photoreceptor changes longitudinally in Stargardt disease.
Subject(s)
Macular Degeneration , Ophthalmoscopy , Retinal Cone Photoreceptor Cells , Retinal Rod Photoreceptor Cells , Stargardt Disease , Tomography, Optical Coherence , Humans , Ophthalmoscopy/methods , Retinal Cone Photoreceptor Cells/pathology , Male , Tomography, Optical Coherence/methods , Retinal Rod Photoreceptor Cells/pathology , Macular Degeneration/congenital , Macular Degeneration/diagnosis , Macular Degeneration/genetics , Visual Acuity , Adult , Fluorescein Angiography/methods , Cell Count , AdolescentABSTRACT
BACKGROUND/AIMS: Mutations in ABCA4 cause Stargardt macular degeneration, which invariably ends in legal blindness. We studied two common mutants, A1038V (in NBD1) and G1961E (in NBD2), with the purpose of exploring how they interact with the cell's quality control mechanism. The study was designed to determine how these mutants can be rescued. METHODS: We expressed wt and mutant ABCA4 in HEK293 cells and studied the effect of the mutations on trafficking and processing and the ability of correctors to rescue them. We used a combination of western blotting, confocal microscopy and surface biotinylation coupled with pulldown of plasma membrane proteins. RESULTS: G1961E is sensitive to inhibitors of the aggresome, tubacin and the lysosome, bafilomycin A. Both mutants cause a reduction in heat shock protein, Hsp27. Incubation of HEK293 cells expressing the mutants with VX-809, an FDA approved drug for the treatment of cystic fibrosis, increased the levels of A1038V and G1961E by 2- to 3-fold. Importantly, VX-809 increased the levels of both mutants at the plasma membrane suggesting that trafficking had been restored. Transfecting additional Hsp27 to the cells also increased the steady state levels of both mutants. However, in combination with VX-809 the addition of Hsp27 caused a dramatic increase in the protein expression particularly in the G1961 mutant which increased approximately 5-fold. CONCLUSION: Our results provide a new mechanism for the rescue of ABCA4 trafficking mutants based on the restoration of Hsp27. Our results provide a pathway for the treatment of Stargardt disease.
Subject(s)
ATP-Binding Cassette Transporters/metabolism , Aminopyridines/pharmacology , Benzodioxoles/pharmacology , ATP-Binding Cassette Transporters/genetics , Aminopyridines/therapeutic use , Anilides/pharmacology , Benzodioxoles/therapeutic use , Cell Membrane/metabolism , Cystic Fibrosis Transmembrane Conductance Regulator/genetics , Cystic Fibrosis Transmembrane Conductance Regulator/metabolism , Gene Expression Regulation/drug effects , HEK293 Cells , HSP27 Heat-Shock Proteins/metabolism , Humans , Hydroxamic Acids/pharmacology , Leupeptins/pharmacology , Lysosomes/metabolism , Macular Degeneration/congenital , Macular Degeneration/drug therapy , Macular Degeneration/metabolism , Macular Degeneration/pathology , Mutation , Protein Transport/drug effects , Stargardt DiseaseABSTRACT
Objective: Microperimetry (MP) is used to assess visual sensitivity mediated by the central retina. As such, MP performance is a candidate outcome measure for gene therapy trials. Herein, we review MP results in three inherited retinal disorders for which gene therapy trials have been initiated-choroideremia, Stargardt disease, and X-linked juvenile retinoschisis. Each of these disorders typically presents in childhood and each has distinct effects on the central retina. Outcomes and Results: Our review indicates that microperimetry is feasible in each of these conditions. The MP sensitivity maps vary among conditions consistent with known effects of each of the three conditions. There is, however, within each of the three disorders considerable variability in fixation stability and in the pattern of sensitivity loss. Conclusions: Microperimetry is a valuable tool for monitoring functional aspects of central retina in an individual patient, especially in combination with other modalities such as OCT, autofluorescence, and acuity and thus may contribute to evaluating the efficacy of gene treatments. Variability of the MP parameters raises some cautions in application of MP as an outcome measure in treatment trials that may have small sample sizes. Nonetheless, we suspect that MP will continue to have a rightful place in future gene therapy trials.
Subject(s)
Choroideremia/physiopathology , Macular Degeneration/congenital , Retinoschisis/physiopathology , Visual Field Tests/methods , Visual Fields/physiology , Choroideremia/diagnosis , Clinical Trials as Topic , Fixation, Ocular/physiology , Humans , Macular Degeneration/diagnosis , Macular Degeneration/physiopathology , Retinoschisis/diagnosis , Stargardt DiseaseABSTRACT
PURPOSE: To raise awareness of Stargardt disease (STGD1) patients without fundus abnormalities. METHODS: Medical records were evaluated for age at onset, initial symptoms and diagnosis, reason for delay of diagnosis, age at STGD1 diagnosis, best-corrected visual acuity (BCVA), ophthalmoscopy, fundus photography, fundus autofluorescence (FAF), fluorescein angiography (FA), spectral-domain optical coherence tomography (SD-OCT), full-field electroretinography (ffERG), color vision test, and the presence of ABCA4 variants. RESULTS: In 11.1% of our STGD1 cohort of 280 patients, no fundus abnormalities were observed at first ophthalmic consultation. The median age at onset was 8 years (range, 1-18). There was a median delay in diagnosis of 3 years (range, 0-19) in 27 out of 31 patients, which resulted in a median age at diagnosis of 12 years (range, 7-26). Patients were misdiagnosed with amblyopia, myopia, optic disk pathology, mental health problems, tension headache, neuritis bulbaris, and uveitis. Subtle abnormalities, such as lipofuscin accumulation, were seen on FAF at an earlier disease stage than in ophthalmoscopy. On SD-OCT, this included a thickened external limiting membrane. Color vision tests showed red-green insufficiency in 79% of patients. Reduced ERG amplitudes were only present in 26% (N = 8) and a dark choroid sign in 65% of the patients. Visual acuity considerably fluctuated in the first 5 years after onset. The majority of the patients (65%) carried a least one variant with a severe effect on ABCA4 function. CONCLUSIONS: Childhood-onset STGD1 patients were diagnosed with a delay of median 3 years. The presence of accurate competence, equipment, and the possibility for genetic screening is required; therefore, we recommend to refer children with visual complaints without initial fundus abnormalities to a specialized ophthalmologic center. In particular, to diagnose patients at an early stage of disease is of increased importance with the advent of new therapeutic possibilities.
Subject(s)
Electroretinography/methods , Fluorescein Angiography/methods , Macular Degeneration/congenital , Retinal Pigment Epithelium/pathology , Tomography, Optical Coherence/methods , Visual Acuity , ATP-Binding Cassette Transporters/genetics , ATP-Binding Cassette Transporters/metabolism , Adolescent , Adult , Child , Child, Preschool , Diagnostic Errors , Female , Fundus Oculi , Genetic Testing , Humans , Infant , Macular Degeneration/diagnosis , Macular Degeneration/genetics , Macular Degeneration/metabolism , Male , Ophthalmoscopy , Stargardt Disease , Young AdultABSTRACT
Autosomal recessive retinal degenerative diseases cause visual impairment and blindness in both humans and dogs. Currently, no standard treatment is available, but pioneering gene therapy-based canine models have been instrumental for clinical trials in humans. To study a novel form of retinal degeneration in Labrador retriever dogs with clinical signs indicating cone and rod degeneration, we used whole-genome sequencing of an affected sib-pair and their unaffected parents. A frameshift insertion in the ATP binding cassette subfamily A member 4 (ABCA4) gene (c.4176insC), leading to a premature stop codon in exon 28 (p.F1393Lfs*1395), was identified. In contrast to unaffected dogs, no full-length ABCA4 protein was detected in the retina of an affected dog. The ABCA4 gene encodes a membrane transporter protein localized in the outer segments of rod and cone photoreceptors. In humans, the ABCA4 gene is associated with Stargardt disease (STGD), an autosomal recessive retinal degeneration leading to central visual impairment. A hallmark of STGD is the accumulation of lipofuscin deposits in the retinal pigment epithelium (RPE). The discovery of a canine homozygous ABCA4 loss-of-function mutation may advance the development of dog as a large animal model for human STGD.
Subject(s)
ATP Binding Cassette Transporter, Subfamily A, Member 4/genetics , Dog Diseases/genetics , Macular Degeneration/congenital , Mutation , ATP Binding Cassette Transporter, Subfamily A, Member 4/chemistry , ATP Binding Cassette Transporter, Subfamily A, Member 4/metabolism , ATP-Binding Cassette Transporters/genetics , Amino Acid Sequence , Animals , Base Sequence , Codon, Nonsense , Disease Models, Animal , Dog Diseases/metabolism , Dog Diseases/pathology , Dogs , Female , Genes, Recessive , Homozygote , Humans , Lipofuscin/metabolism , Macular Degeneration/genetics , Macular Degeneration/metabolism , Macular Degeneration/veterinary , Male , Microscopy, Fluorescence , Models, Molecular , Mutagenesis, Insertional , Pedigree , Protein Conformation , Retina/metabolism , Retina/pathology , Stargardt Disease , Whole Genome SequencingABSTRACT
Purpose: Inherited retinal diseases (IRDs) are clinically and genetically heterogeneous showing progressive retinal cell death which results in vision loss. IRDs include a wide spectrum of disorders, such as retinitis pigmentosa (RP), Leber congenital amaurosis (LCA), cone-rod dystrophy (CRD), and Stargardt disease (STGD1). Methods: In this study, we performed targeted next-generation sequencing based on molecular inversion probes (MIPs) that allowed the sequence analysis of 108 IRD-associated genes in 50 Iranian IRD probands. Results: The sequencing and variant filtering led to the identification of putative pathogenic variants in 36 out of 50 (72%) probands. Among 36 unique variants, we identified 20 novel variants in 15 genes. Four out of 36 probands carry compound heterozygous variants, and 32 probands carry homozygous variants. Conclusions: Employing a cost-effective targeted next-generation sequencing procedure, we identified the genetic causes of different retinal disorders in the majority of Iranian families in this study.
Subject(s)
Cone-Rod Dystrophies/genetics , Eye Proteins/genetics , Leber Congenital Amaurosis/genetics , Macular Degeneration/congenital , Mutation , Retinitis Pigmentosa/genetics , Adolescent , Adult , Child , Cone-Rod Dystrophies/metabolism , Cone-Rod Dystrophies/pathology , Eye Proteins/metabolism , Female , Gene Expression , Genetic Association Studies , Genotype , Heterozygote , High-Throughput Nucleotide Sequencing/economics , High-Throughput Nucleotide Sequencing/methods , Homozygote , Humans , Iran , Leber Congenital Amaurosis/metabolism , Leber Congenital Amaurosis/pathology , Macular Degeneration/genetics , Macular Degeneration/metabolism , Macular Degeneration/pathology , Male , Pedigree , Phenotype , Retina/metabolism , Retina/pathology , Retinitis Pigmentosa/congenital , Retinitis Pigmentosa/metabolism , Retinitis Pigmentosa/pathology , Stargardt DiseaseABSTRACT
We report the generation of the human iPSC line LEIi007-A from a patient with autosomal recessive Stargardt disease caused by compound heterozygous mutations in the ABCA4 gene (c.[5461-10â¯Tâ¯>â¯C];[4139Câ¯>â¯T]). Reprogramming of patient dermal fibroblasts was performed using episomal plasmids containing OCT4, SOX2, KLF4, L-MYC, LIN28, shRNA for p53 and mir302/367 microRNA to establish the clonal iPSC line LEIl007-A. LEIl007-A displayed normal pluripotent stem cell colony morphology, expressed pluripotent stem cell markers, displayed a normal karyotype and differentiated into ectodermal, mesodermal and endodermal germ layer lineages.
Subject(s)
ATP-Binding Cassette Transporters/genetics , Cell Culture Techniques/methods , Genes, Recessive , Induced Pluripotent Stem Cells/pathology , Macular Degeneration/congenital , Adolescent , Base Sequence , Cell Line , Child , Female , Humans , Kruppel-Like Factor 4 , Macular Degeneration/genetics , Stargardt DiseaseABSTRACT
PURPOSE: Automated spectral domain optical coherence tomography (SD-OCT) segmentation algorithms currently do not perform well in segmenting individual intraretinal layers in eyes with Stargardt disease (STGD). We compared selective B-scan segmentation strategies for generating mean retinal layer thickness and preserved area data from SD-OCT scans in patients with STGD1. METHODS: Forty-five eyes from 40 Stargardt patients were randomly selected from the ongoing Natural History of the Progression of Atrophy Secondary to Stargardt Disease (ProgStar) study. All eyes underwent SD-OCT using a standard macular volume consisting of 1024 × 49 equally spaced B-scans within a 20 × 20 degree field centered on the fovea. All 49 B-scans were segmented manually to quantify total retina, outer nuclear layer (ONL), photoreceptor inner segments, photoreceptor outer segments (OS), and retinal pigment epithelial layer (RPE). Mean thickness and total area were generated using all 49 B-scans (spaced 122 µm apart), 25 B-scans (every other B-scan, spaced 240 µm apart), 17 B-scans (every third scan, 353 µm apart), and 13 B-scans (every fourth scan, 462 µm apart), as well as by using an "adaptive" method where a subset (minimum 25 B-scans) of B-scans that the grader deemed as significantly different from adjacent B-scans were utilized. Mean absolute and percentage errors were calculated for macular thickness and area of different retinal layers for the different B-scan subset selection strategies relative to using all 49 B-scans, which was considered the reference or ground truth. RESULTS: Mean thickness and area measurements were significantly different for any regularly spaced reduction in B-scan density relative to the ground truth. When an adaptive approach was applied using a minimum of half the scans, the differences relative to ground truth were no longer significantly different. The mean percent differences for the area and thicknesses of the various layers ranged from 0.02 to 33.66 (p < 0.05 for all comparisons) and 0.44 to 7.24 (p > 0.05) respectively. CONCLUSION: Manual segmentation of a subset of B-scans using an adaptive strategy can yield thickness and area measurements of retinal sublayers comparable to the reference ground truth derived from using all B-scans in the volume. These results may have implications for increasing the efficiency of SD-OCT grading strategies in clinical trials for STGD and other related macular degenerative disorders.
Subject(s)
Algorithms , Macula Lutea/pathology , Macular Degeneration/congenital , Retinal Pigment Epithelium/pathology , Tomography, Optical Coherence/methods , Follow-Up Studies , Humans , Macular Degeneration/diagnosis , Prospective Studies , Reproducibility of Results , Severity of Illness Index , Stargardt DiseaseABSTRACT
Stargardt disease (STGD1, OMIM 248200) is a common hereditary juvenile or early adult onset macular degeneration. It ultimately leads to progressive central vision loss. Here, we sought to identify gene mutations associated with STGD1 in a three-generation Han Chinese pedigree by whole exome sequencing and Sanger sequencing. Two novel potentially pathogenic variants in a compound heterozygous state, c.3607G>T (p.(Gly1203Trp)) and c.6722T>C (p.(Leu2241Pro)), in the ATP binding cassette subfamily A member 4 gene (ABCA4) were identified as contributing to the family's STGD1 phenotype. These variants may impact the ABCA4 protein structure and reduce the retinal-activated ATPase activity, leading to abnormal all-trans retinal accumulation in photoreceptor outer segments and in retinal pigment epithelium cells. The present study broadens the mutational spectrum of the ABCA4 responsible for STGD1. A combination of whole exome sequencing and Sanger sequencing is likely to be a time-saving and cost-efficient approach to screen pathogenic variants in genetic disorders caused by sizable genes, as well as avoiding misdiagnosis. These results perhaps refine genetic counseling and ABCA4-targetted treatments for families affected by STGD1.
Subject(s)
ATP-Binding Cassette Transporters/genetics , Macular Degeneration/congenital , Polymorphism, Genetic , Retinal Pigment Epithelium/metabolism , ATP-Binding Cassette Transporters/chemistry , ATP-Binding Cassette Transporters/metabolism , Adult , Amino Acid Sequence , Animals , Asian People , Base Sequence , Case-Control Studies , Female , Gene Expression , Heterozygote , Humans , Macular Degeneration/ethnology , Macular Degeneration/genetics , Macular Degeneration/metabolism , Macular Degeneration/pathology , Male , Models, Molecular , Pedigree , Phenotype , Protein Conformation, alpha-Helical , Protein Conformation, beta-Strand , Retinal Pigment Epithelium/pathology , Sequence Alignment , Sequence Homology, Amino Acid , Stargardt Disease , Exome SequencingABSTRACT
PURPOSE: The present study describes retinotopic mapping of the primary visual cortex using functional MRI (fMRI) in patients with retinal disease. It addresses the relationship between fMRI data and data obtained by conventional assessment including microperimetry (MP) and structural imaging. METHODS: Initial testing involved eight patients with central retinal disease (Stargardt disease, STGD) and eight with peripheral retinal disease (retinitis pigmentosa, RP), who were examined using fMRI and MP (Nidek MP-1). All had a secure clinical diagnosis supported by electrophysiological data. fMRI used population-receptive field (pRF) mapping to provide retinotopic data that were then compared with the results of MP, optical coherence tomography and fundus autofluorescence imaging. RESULTS: Full analysis, following assessment of fMRI data reliability criteria, was performed in five patients with STGD and seven patients with RP; unstable fixation was responsible for unreliable pRF measurements in three patients excluded from final analysis. The macular regions in patients with STGD with central visual field defects and outer retinal atrophy (ORA) at the macula correlated well with pRF coverage maps showing reduced density of activated voxels at the occipital pole. Patients with RP exhibited peripheral ORA and concentric visual field defects both on MP and pRF mapping. Anterior V1 voxels, corresponding to peripheral regions, showed no significant activation. Correspondence between MP and pRF mapping was quantified by calculating the simple matching coefficient. CONCLUSION: Retinotopic maps acquired by fMRI provide a valuable adjunct in the assessment of retinal dysfunction. The addition of microperimetric data to pRF maps allowed better assessment of macular function than MP alone. Unlike MP, pRF mapping provides objective data independent of psychophysical perception from the patient.
Subject(s)
Macular Degeneration/congenital , Retinitis Pigmentosa/diagnostic imaging , Visual Cortex/diagnostic imaging , Adolescent , Adult , Brain Mapping , Female , Humans , Macular Degeneration/diagnostic imaging , Magnetic Resonance Imaging , Male , Middle Aged , Scotoma/diagnostic imaging , Stargardt Disease , Tomography, Optical Coherence , Visual Field Tests , Visual Fields/physiology , Young AdultABSTRACT
BACKGROUND/AIMS: To describe the genetic characteristics of the cohort enrolled in the international multicentre progression of Stargardt disease 1 (STGD1) studies (ProgStar) and to determine geographic differences based on the allele frequency. METHODS: 345 participants with a clinical diagnosis of STGD1 and harbouring at least one disease-causing ABCA4 variant were enrolled from 9 centres in the USA and Europe. All variants were reviewed and in silico analysis was performed including allele frequency in public databases and pathogenicity predictions. Participants with multiple likely pathogenic variants were classified into four national subgroups (USA, UK, France, Germany), with subsequent comparison analysis of the allele frequency for each prevalent allele. RESULTS: 211 likely pathogenic variants were identified in the total cohort, including missense (63%), splice site alteration (18%), stop (9%) and others. 50 variants were novel. Exclusively missense variants were detected in 139 (50%) of 279 patients with multiple pathogenic variants. The three most prevalent variants of these patients with multiple pathogenic variants were p.G1961E (15%), p.G863A (7%) and c.5461-10 T>C (5%). Subgroup analysis revealed a statistically significant difference between the four recruiting nations in the allele frequency of nine variants. CONCLUSIONS: There is a large spectrum of ABCA4 sequence variants, including 50 novel variants, in a well-characterised cohort thereby further adding to the unique allelic heterogeneity in STGD1. Approximately half of the cohort harbours missense variants only, indicating a relatively mild phenotype of the ProgStar cohort. There are significant differences in allele frequencies between nations, although the three most prevalent variants are shared as frequent variants.
Subject(s)
ATP-Binding Cassette Transporters/genetics , Macular Degeneration/congenital , Mutation , Adolescent , Adult , Age of Onset , Aged , Child , Child, Preschool , Cohort Studies , DNA Mutational Analysis , Databases, Factual , Female , Gene Frequency , Genotyping Techniques , Geography , Humans , Internationality , Macular Degeneration/diagnosis , Macular Degeneration/genetics , Male , Middle Aged , Polymerase Chain Reaction , Stargardt DiseaseABSTRACT
PURPOSE: To investigate genetic mutations in Korean patients with Stargardt disease (STGD) using exome sequencing, and to analyze the correlations between genetic mutations and clinical phenotypes. METHODS: Peripheral venous blood was obtained from 24 clinically diagnosed Korean STGD patients, followed by extraction of genomic DNAs. Using exome sequencing we investigated gene mutations for the adenosine triphosphate-binding cassette, subfamily A, member 4 (ABCA4) elongation of very-long-chain fatty acids 4 (ELOVL4), and prominin 1 (PROM1), and confirmed gene mutations by the direct sequencing of polymerase chain reaction products. RESULTS: ABCA4 mutations were confirmed in 17 of 24 patients, and 12 novel mutations were identified. ELOVL4 and PROM1 gene mutations were not identified in this study. We also identified 16 previously reported mutations related to STGD1. In patients whose disease symptoms occurred before 20 years of age, visual acuity was poorer and atrophic flecks were more frequently found. In addition, more ABCA4 mutations were found in patients who had choroidal silence or atrophic flecks. CONCLUSIONS: Novel ABCA4 gene mutations were found in Korean patients with STGD1. This study will facilitate better understanding of the relationships between ABCA4 gene mutations and clinical symptoms in Korean patients.
Subject(s)
ATP-Binding Cassette Transporters/genetics , DNA/genetics , Genetic Testing/methods , Macular Degeneration/congenital , Mutation , ATP-Binding Cassette Transporters/metabolism , Adult , Aged , DNA Mutational Analysis , Electroretinography , Exome , Female , Fluorescein Angiography , Fundus Oculi , Humans , Incidence , Macular Degeneration/diagnosis , Macular Degeneration/epidemiology , Macular Degeneration/genetics , Male , Middle Aged , Pedigree , Phenotype , Polymerase Chain Reaction , Republic of Korea/epidemiology , Retina/pathology , Retrospective Studies , Rod Cell Outer Segment , Stargardt Disease , Young AdultABSTRACT
PURPOSE: To describe the hyperreflective foci (HF) characteristics in eyes affected by Stargardt disease (STGD), correlating HF with the atrophy progression at 1-year follow-up. METHODS: Prospective observational case series with 1-year follow-up. Twenty-eight eyes (14 patients) affected by STGD and 28 eyes (14 age- and sex-matched healthy patients) used as control group were recruited. All patients underwent a complete ophthalmologic examination including fundus autofluorescence and spectral-domain optical coherence tomography. The primary outcome was the identification of HF specific location in STGD and their modification over a 1-year follow-up. Secondary outcome included the correlation between the number and the location of HF and atrophic changes. RESULTS: HF turned out to be more frequent in STGD patients compared with healthy controls (p < 0.001). In particular, mean number of HF in the pathological edge was significantly higher than in the healthy edge of the atrophy (p < 0.001) and in the foveal area (p < 0.001). A negative correlation was found between the total HF number in the pathological edge and the atrophic area at baseline. HF number in the outer retina of the pathological edge significantly decreased between the baseline and the final follow-up examination (p = 0.011). The enlargement of the atrophic area in eyes with more than five outer retinal HF in the pathological edge at baseline was significantly less than that found in the eyes with fewer than five HF (p = 0.010). CONCLUSIONS: HF are most common at the pathological margin of the central atrophy, with outer retina foci being more frequently found in smaller atrophic lesions.
Subject(s)
Fluorescein Angiography/methods , Macular Degeneration/congenital , Retina/pathology , Tomography, Optical Coherence/methods , Visual Acuity , Adult , Aged , Disease Progression , Female , Follow-Up Studies , Fundus Oculi , Humans , Macular Degeneration/diagnosis , Male , Middle Aged , Prospective Studies , Stargardt Disease , Time Factors , Young AdultABSTRACT
PURPOSE: To describe the study design and characteristics at first visit of participants in the longitudinal Scotopic Microperimetric Assessment of Rod Function in Stargardt Disease (SMART) study. METHODS: Scotopic microperimetry (sMP) was performed in one designated study eye in a subset of participants with molecularly proven ABCA4-associated Stargardt disease (STGD1) enrolled in a multicenter natural history study (ProgStar). Study visits were every 6 months over a period ranging from 6 to 24 months, and also included fundus autofluorescence (FAF). RESULTS: SMART enrolled 118 participants (118 eyes). At the first visit of SMART, the mean sensitivity in mesopic microperimetry was 11.48 (±5.05; range 0.00-19.88) dB and in sMP 11.25 (±5.26; 0-19.25) dB. For FAF, all eyes had a lesion of decreased autofluorescence (mean lesion size 3.62 [±3.48; 0.10-21.46] mm2), and a total of 76 eyes (65.5%) had a lesion of definitely decreased autofluorescence with a mean lesion size of 3.46 (±3.60; 0.21-21.46) mm2. CONCLUSIONS: Rod function is impaired in STGD1 and can be assessed by sMP. Testing rod function may serve as a potential outcome measure for future clinical treatment trials. This is evaluated in the SMART study.
Subject(s)
Macular Degeneration/congenital , Night Vision/physiology , Retinal Rod Photoreceptor Cells/physiology , Visual Fields/physiology , Adult , Aged , Female , Humans , Longitudinal Studies , Macular Degeneration/physiopathology , Male , Middle Aged , Research Design , Stargardt Disease , Visual Acuity/physiology , Visual Field Tests , Young AdultABSTRACT
Stargardt disease (STGD) is one of the most common macular dystrophies in young adults. It progresses slowly. Its prevalence is about 1:8000-10,000. Age of onset is a surrogate marker: The earlier the onset, the more severe the disease course. Onset usually occurs in childhood or early adolescence, at about 10-15 years of age. Vision is between about 20/70 and 20/200. The fundus shows a bull's eye pattern or beaten-bronze appearance, with or without yellowish flecks (fundus flavimaculatus). Fluorescein angiography may show dark choroid in about 80% of cases. On fundus autofluorescence (FAF), newer flecks appear hyperautofluorescent (hyperAF); older ones become progressively more hypoAF with time. Some flecks are surrounded by a ring of decreased AF. Peripapillary sparing is one the characteristics of Stargardt disease, but this area can be involved in about 2-7% of cases. The reason for this sparing is unclear; this area may be more resilient to the deleterious effect of ABCA4 gene mutation, and there might be a more favorable RPE photoreceptor ratio, resulting in less lipofuscin build-up, in the presence of a thicker overlying peripapillary retinal nerve fiber layer. Patients with Stargardt disease should avoid bright light and excessive vitamin A.
Subject(s)
Macular Degeneration/congenital , ATP-Binding Cassette Transporters/genetics , Adolescent , Child , Fluorescein Angiography , Fundus Oculi , Humans , Macular Degeneration/physiopathology , Stargardt DiseaseABSTRACT
Macular degeneration is hallmarked by retinal accumulation of toxic retinoid species (e.g., A2E) for which there is no endogenous mechanism to eliminate it. This ultimately results in progressive dysfunction and loss of vision either in advanced age for genetically normal patients (age-related macular degeneration) or in adolescence for those with inherited genetic mutations (Stargardt's disease). In this article, we present a proof-of-concept study for an enzyme-based therapy to remove these retinoids, modeled on traditional enzyme replacement therapy. Recombinant manganese peroxidase (rMnP) is produced in Pichia pastoris. In vitro, we demonstrate that rMnP breaks down A2E and other lipofuscin fluorophores with limited cellular toxicity, and as this enzyme is mannosylated, it can be taken up into cells through mannose receptor-dependent endocytosis. In vivo, we demonstrate that rMnP can significantly reduce the A2E burden when administered by intravitreal injections. Together, these data provide encouraging results toward the development of an enzyme-based therapy for macular degeneration and indicate the need for additional work to characterize the molecular mechanism of A2E breakdown and to improve the pharmacological parameters of the enzyme.