Anne B. Fulton

Identification of the gene (BBS1) most commonly involved in Bardet-Biedl syndrome, a complex human obesity syndrome.

Bardet-Biedl syndrome (BBS, OMIM 209900) is a genetic disorder with the primary features of obesity, pigmentary retinopathy, polydactyly, renal malformations, mental retardation and hypogenitalism. Individuals with BBS are also at increased risk for diabetes mellitus, hypertension and congenital heart disease. What was once thought to be a homogeneous autosomal recessive disorder is now known to map to at least six loci: 11q13 (BBS1), 16q21 (BBS2), 3p13–p12 (BBS3), 15q22.3–q23 (BBS4), 2q31 (BBS5) and 20p12 (BBS6). There has been considerable interest in identifying the genes that underlie BBS, because some components of the phenotype are common. Cases of BBS mapping ro BBS6 are caused by mutations in MKKS; mutations in this gene also cause McKusick-Kaufman syndrome (hydrometrocolpos, post-axial polydactyly and congenital heart defects). In addition, we recently used positional cloning to identify the genes underlying BBS2 (ref. 16) and BBS4 (ref. 17). The BBS6 protein has similarity to a Thermoplasma acidophilum chaperonin, whereas BBS2 and BBS4 have no significant similarity to chaperonins. It has recently been suggested that three mutated alleles (two at one locus, and a third at a second locus) may be required for manifestation of BBS (triallelic inheritance). Here we report the identification of the gene BBS1 and show that a missense mutation of this gene is a frequent cause of BBS. In addition, we provide data showing that this common mutation is not involved in triallelic inheritance.

The human rod ERG: Correlation with psychophysical responses in light and dark adaptation

Abstract The a- and b-waves over a large range of human scotopic ERG responses in various states of adaptation may be represented compactly by “H2” curves. This representation allows correlation of b-wave and psychophysical thresholds obtained in the same conditions of adaptation and shows sensitivity and scaling of response size are two separable features of both the a- and b-waves in light and dark adaptation. The relation of these responses to the underlying retinal processes is discussed.

Foveal Fine Structure in Retinopathy of Prematurity: An Adaptive Optics Fourier Domain Optical Coherence Tomography Study

PURPOSE To describe the fine structure of the fovea in subjects with a history of mild retinopathy of prematurity (ROP) using adaptive optics-Fourier domain optical coherence tomography (AO-FDOCT). METHODS High-speed, high-resolution AO-FDOCT videos were recorded in subjects with a history of ROP (n = 5; age range, 14-26 years) and in control subjects (n = 5; age range, 18-25 years). Custom software was used to extract foveal pit depth and volume from three-dimensional (3-D) retinal maps. The thickness of retinal layers as a function of retinal eccentricity was measured manually. The retinal vasculature in the parafoveal region was assessed. RESULTS The foveal pit was wider and shallower in ROP than in control subjects. Mean pit depth, defined from the base to the level at which the pit reaches a lateral radius of 728 microm, was 121 microm compared with 53 microm. Intact, contiguous inner retinal layers overlay the fovea in ROP subjects but were absent in the control subjects. Mean full retinal thickness at the fovea was greater in the subjects with ROP (279.0 microm vs. 190.2 microm). The photoreceptor layer thickness did not differ between ROP and control subjects. An avascular zone was not identified in the subjects with ROP but was present in all the control subjects. CONCLUSIONS The foveas of subjects with a history of mild ROP have significant structural abnormalities that are probably a consequence of perturbations of neurovascular development.

Cycloplegic Refractions in Infants and Young Children

We studied groups of normal infants and infants with amblyopia and esoropia to determine the incidence of infantile astigmatism. Under cycloplegia, 19% of normal infants had astigmatism; this was at least twice the incidence in adults, but less than one-hale that found by noncycloplegic refractions of infants. During the first three postnatal years the incidence of astigmatism and distributions of spherical equivalents and anisometropia did not distinguish normal patients from most of those with esotropia and amblyopia.

Evaluation of Complex Inheritance Involving the Most Common Bardet-Biedl Syndrome Locus (BBS1)

Bardet-Biedl syndrome (BBS) is a genetic disorder with the primary features of obesity, pigmentary retinopathy, polydactyly, renal malformations, mental retardation, and hypogenitalism. Patients with BBS are also at increased risk for diabetes mellitus, hypertension, and congenital heart disease. BBS is known to map to at least six loci: 11q13 (BBS1), 16q21 (BBS2), 3p13-p12 (BBS3), 15q22.3-q23 (BBS4), 2q31 (BBS5), and 20p12 (BBS6). Although these loci were all mapped on the basis of an autosomal recessive mode of inheritance, it has recently been suggested-on the basis of mutation analysis of the identified BBS2, BBS4, and BBS6 genes-that BBS displays a complex mode of inheritance in which, in some families, three mutations at two loci are necessary to manifest the disease phenotype. We recently identified BBS1, the gene most commonly involved in Bardet-Biedl syndrome. The identification of this gene allows for further evaluation of complex inheritance. In the present study we evaluate the involvement of the BBS1 gene in a cohort of 129 probands with BBS and report 10 novel BBS1 mutations. We demonstrate that a common BBS1 missense mutation accounts for approximately 80% of all BBS1 mutations and is found on a similar genetic background across populations. We show that the BBS1 gene is highly conserved between mice and humans. Finally, we demonstrate that BBS1 is inherited in an autosomal recessive manner and is rarely, if ever, involved in complex inheritance.

Panel-based Genetic Diagnostic Testing for Inherited Eye Diseases is Highly Accurate and Reproducible and More Sensitive for Variant Detection Than Exome Sequencing

Purpose:Next-generation sequencing–based methods are being adopted broadly for genetic diagnostic testing, but the performance characteristics of these techniques with regard to test accuracy and reproducibility have not been fully defined.Methods:We developed a targeted enrichment and next-generation sequencing approach for genetic diagnostic testing of patients with inherited eye disorders, including inherited retinal degenerations, optic atrophy, and glaucoma. In preparation for providing this genetic eye disease (GEDi) test on a CLIA–certified basis, we performed experiments to measure the sensitivity, specificity, and reproducibility, as well as the clinical sensitivity, of the test.Results:The GEDi test is highly reproducible and accurate, with sensitivity and specificity of 97.9 and 100%, respectively, for single-nucleotide variant detection. The sensitivity for variant detection was notably better than the 88.3% achieved by whole-exome sequencing using the same metrics, because of better coverage of targeted genes in the GEDi test as compared with a commercially available exome capture set. Prospective testing of 192 patients with inherited retinal degenerations indicated that the clinical sensitivity of the GEDi test is high, with a diagnostic rate of 51%.Conclusion:Based on quantified performance metrics, the data suggest that selective targeted enrichment is preferable to whole-exome sequencing for genetic diagnostic testing.Genet Med 17 4, 253–261.

The Relation of Myopia and Astigmatism in Developing Eyes

The relation of astigmatism and myopia was analyzed in 298 myopic children, ages birth to 10 years. The mean spherical equivalent, determined by cyclopentolate retinoscopy, for the entire group was--2.9 diopters and did not change significantly with age. However, in 3-year-old children and younger, myopia progressed in eyes with greater than or equal to 1 diopter of cylinder and tended to increase through age 8 years in those having greater than or equal to 3 diopters of cylinder. Also, astigmatic errors greater than or equal to 1 diopter, especially of oblique orientation, were associated with higher degrees of myopia than nonastigmatic errors. These data from myopic children suggest that uncorrected astigmatism during a period of visual immaturity influences the course of myopia. Thus, naturally occurring astigmatic errors, that are frequent among infants and young children, appear to have a role similar to the vision blurring perturbations that trigger the development of myopia in young animals. Ascertainment and full correction of these refractive errors in young children may be important in assuring the best possible vision.

The Neurovascular Retina in Retinopathy of Prematurity

The continuing worldwide epidemic of retinopathy of prematurity (ROP), a leading cause of childhood visual impairment, strongly motivates further research into mechanisms of the disease. Although the hallmark of ROP is abnormal retinal vasculature, a growing body of evidence supports a critical role for the neural retina in the ROP disease process. The age of onset of ROP coincides with the rapid developmental increase in rod photoreceptor outer segment length and rhodopsin content of the retina with escalation of energy demands. Using a combination of non-invasive electroretinographic (ERG), psychophysical, and image analysis procedures, the neural retina and its vasculature have been studied in prematurely born human subjects, both with and without ROP, and in rats that model the key vascular and neural parameters found in human ROP subjects. These data are compared to comprehensive numeric summaries of the neural and vascular features in normally developing human and rat retina. In rats, biochemical, anatomical, and molecular biological investigations are paired with the non-invasive assessments. ROP, even if mild, primarily and persistently alters the structure and function of photoreceptors. Post-receptor neurons and retinal vasculature, which are intimately related, are also affected by ROP; conspicuous neurovascular abnormalities disappear, but subtle structural anomalies and functional deficits may persist years after clinical ROP resolves. The data from human subjects and rat models identify photoreceptor and post-receptor targets for interventions that promise improved outcomes for children at risk for ROP.

Development of ERG responses: The ISCEV rod, maximal and cone responses in normal subjects

Purpose: Summarize ISCEV ERG responses from normal infants and children. Methods: The amplitudes and implicit times of the ISCEV rod, maximal dark-adapted and cone responses from a total of 409 normal infants (n=128), children and adult controls were compiled. The subjects, aged 1 week to 52 years, were divided into seven age groups, including four in infancy (<52 weeks). The response parameters for each age group were summarized as percentiles. Results: In each ISCEV condition, the youngest infants (1–5 weeks) had significantly smaller amplitudes and longer implicit times than adults. Amplitude increased and implicit time decreased systematically with age. Conclusions: The developmental changes in ERG responses are significant. The medians and ranges herein provide provisional norms against which the ERG responses from pediatric patients can be compared

Photoreceptor function in infants and children with a history of mild retinopathy of prematurity.

Five infants and children with a history of mild retinopathy of prematurity (ROP) were tested for postulated alterations in rod photoreceptor function. The photoreceptor responses were derived from the electroretinographic alpha waves. Postreceptoral components, the beta wave and the oscillatory potentials, were also examined. The saturated amplitude and sensitivity of the rod photoreceptor responses were low, except for the sensitivity in one patient. The beta-wave sensitivity was low, but saturated amplitudes were within the 95% prediction interval for normal. The amplitudes of the oscillatory-potential responses were also attenuated. The results indicate that retinal dysfunction may be present in patients with a history of mild ROP long after the ROP has completely resolved. Additionally, the data suggest that the photoreceptors are the primary site of retinal dysfunction in mild ROP.