Marko Nardini

Long-term effect of gene therapy on Leber's congenital amaurosis.

BACKGROUND Mutations in RPE65 cause Lebers congenital amaurosis, a progressive retinal degenerative disease that severely impairs sight in children. Gene therapy can result in modest improvements in night vision, but knowledge of its efficacy in humans is limited. METHODS We performed a phase 1-2 open-label trial involving 12 participants to evaluate the safety and efficacy of gene therapy with a recombinant adeno-associated virus 2/2 (rAAV2/2) vector carrying the RPE65 complementary DNA, and measured visual function over the course of 3 years. Four participants were administered a lower dose of the vector, and 8 were administered a higher dose. In a parallel study in dogs, we investigated the relationship among vector dose, visual function, and electroretinography (ERG) findings. RESULTS Improvements in retinal sensitivity were evident, to varying extents, in six participants for up to 3 years, peaking at 6 to 12 months after treatment and then declining. No associated improvement in retinal function was detected by means of ERG. Three participants had intraocular inflammation, and two had clinically significant deterioration of visual acuity. The reduction in central retinal thickness varied among participants. In dogs, RPE65 gene therapy with the same vector at lower doses improved vision-guided behavior, but only higher doses resulted in improvements in retinal function that were detectable with the use of ERG. CONCLUSIONS Gene therapy with rAAV2/2 RPE65 vector improved retinal sensitivity, albeit modestly and temporarily. Comparison with the results obtained in the dog model indicates that there is a species difference in the amount of RPE65 required to drive the visual cycle and that the demand for RPE65 in affected persons was not met to the extent required for a durable, robust effect. (Funded by the National Institute for Health Research and others; ClinicalTrials.gov number, NCT00643747.).

Fusion of visual cues is not mandatory in children

Human adults can go beyond the limits of individual sensory systems’ resolutions by integrating multiple estimates (e.g., vision and touch) to reduce uncertainty. Little is known about how this ability develops. Although some multisensory abilities are present from early infancy, it is not until age ≥8 y that children use multiple modalities to reduce sensory uncertainty. Here we show that uncertainty reduction by sensory integration does not emerge until 12 y even within the single modality of vision, in judgments of surface slant based on stereoscopic and texture information. However, adults’ integration of sensory information comes at a cost of losing access to the individual estimates that feed into the integrated percept (“sensory fusion”). By contrast, 6-y-olds do not experience fusion, but are able to keep stereo and texture information separate. This ability enables them to outperform adults when discriminating stimuli in which these information sources conflict. Further, unlike adults, 6-y-olds show speed gains consistent with following the fastest-available single cue. Therefore, whereas the mature visual system is optimized for reducing sensory uncertainty, the developing visual system may be optimized for speed and for detecting sensory conflicts. Such conflicts could provide the error signals needed to learn the relationships between sensory information sources and to recalibrate them while the body is growing.

Infant Hyperopia: Detection, Distribution, Changes and Correlates—Outcomes From the Cambridge Infant Screening Programs

Purpose. To report on two population screening programs designed to detect significant refractive errors in 8308 8- to 9-month-old infants, examine the sequelae of infant hyperopia, and test whether early partial spectacle correction improved visual outcome (strabismus and acuity). The second program also examined whether infant hyperopia was associated with developmental differences across various domains such as language, cognition, attention, and visuomotor competences up to age 7 years. Linked programs in six European countries assessed costs of infant refractive screening. Method. In the first program, screening included an orthoptic examination and isotropic photorefraction, with cycloplegia. In the second program we carried out the same screening procedure without cycloplegia. Hyperopic infants (≥+4 D) were followed up alongside an emmetropic control group, with visual and developmental measures up to age 7 years, and entered a controlled trial of partial spectacle correction. Results. The second program showed that accommodative lag during photorefraction with a target at 75 cm (focus ≥+1.5 D) was a marker for significant hyperopia. In each program, prevalence of significant hyperopia at 9 to 11 months was around 5%; manifest strabismus was 0.3% at 9 months and 1.5 to 2.0% by school age. Infant hyperopia was associated with increased strabismus and poor acuity at 4 years. Spectacle wear by infant hyperopes produced better visual outcome than in uncorrected infants, although an improvement in strabismus was found in the first program only. The corrections did not affect emmetropization to 3.5 years; however, both corrected and uncorrected groups remained more hyperopic than controls in the preschool years. The hyperopic group showed poorer overall performance than controls between 1 and 7 years on visuoperceptual, cognitive, motor, and attention tests, but showed no consistent differences in early language or phonological awareness. Relative cost estimates suggest that refractive screening programs can detect visual problems in infancy at lower overall cost than surveillance in primary care. Conclusions. Photo/videorefraction can successfully screen infants for refractive errors, with visual outcomes improved through early refractive correction. Infant hyperopia is associated with mild delays across many aspects of visuocognitive and visuomotor development. These studies raise the possibility that infant refractive screening can identify not only visual problems, but also potential developmental and learning difficulties.

Infant vision screening predicts failures on motor and cognitive tests up to school age

In a population-based infant vision screening programme, 5295 infants were screened and those with significant refractive errors were followed up. To assess the relationship between the development of vision and other domains, we report a longitudinal study comparing infants with significant hyperopia, identified at age 9 months (‘hyperopes’) with infants with normal refractions (‘controls’). Children are included who completed at each age a broad set of visual, cognitive, motor and language measures taken over a series of follow-up visits up to age 5.5 years. Hyperopes performed significantly worse than controls on the Atkinson Battery of Child Development for Examining Functional Vision at 14 months and 3.5 years and the Henderson Movement Assessment Battery for Children at 3.5 and 5.5 years. The Griffiths Child Development Scales, MacArthur Communicative Development Inventory and British Picture Vocabulary Scales showed no significant differences. Exclusion of those infants who became amblyopic and strabismic did not substantially alter these results, suggesting that the differences between groups were not a consequence of these disorders. These results indicate that early hyperopia is associated with a range of developmental deficits that persist at least to age 5.5 years. These effects are concentrated in visuocognitive and visuomotor domains rather than the linguistic domain.

Cortical vision, MRI and developmental outcome in preterm infants

Objectives: To test two measures of visual cortical function in the first year of life as early markers of functionally significant brain damage in infants born preterm: orientation-reversal visual event-related potentials (OR-VERP) and a behavioural test of cortically controlled visual attention-fixation shifts under competition (FS). Also to examine how these measures relate to (1) perinatal brain insults identified by MRI, and (2) later neurodevelopmental status. Patients and methods: After neonatal and term-age-equivalent MRI, 26 preterm infants (<32 weeks of gestational age, mean 28.1 weeks) were given the OR-VERP and FS tests before 12 months post-term age and a neurodevelopmental assessment (Griffiths Scales) at 2 years. MRI scans examined for parenchymal lesions, intraventricular haemorrhage, ventricular dilatation and diffuse excessive high signal intensity were classified into three categories of severity. Cortical visual test results were compared across these categories and examined as predictors of developmental status at 2 years. Results: 26 infants were studied. 13/25 infants showed significant OR-VERP responses. 12/26 showed normal FS performance. On both tests, the proportion of infants meeting these criteria decreased significantly with MRI severity. As predictors of Griffiths developmental quotient ⩽80, the FS test had a sensitivity of 100%, a specificity of 61%, and positive and negative predictive values of 50% and 100%, respectively; corresponding values for OR-VERP were 86%, 65%, 50% and 92% . Conclusions: Visual cortical tests can provide early indicators of the functional impact of perinatal brain damage in the preterm infant.

Non-cycloplegic refractive screening can identify infants whose visual outcome at 4 years is improved by spectacle correction.

The Second Cambridge Population Infant Vision Screening Programme using the VPR-1 videorefractor without cycloplegia was undertaken in order to identify those infants with refractive errors who were potentially amblyogenic or strabismogenic. Infants identified at eight months were entered into a control trial of treatment with partial spectacle correction and underwent a long-term follow-up that monitored a wide range of visual, visuoperceptual, visuocognitive, visuomotor, linguistic and social development. In the present paper, the authors report on the outcome measures of visual acuity and strabismus. Poor acuity was defined as a best-corrected acuity of 6/12 or worse on crowded letters or 6/9 or worse on single letters, at age 4 years. Acuity was measured in 79 infants who were significantly hyperopic and/or anisometropic at 11-12 months of age, 23 who showed hyperopia of +3D but less than +3.5D, 196 control subjects, 14 controls with refractive errors, and 126 others who showed an accommodative lag on screening but were not significantly hyperopic on first retinoscopy. There was a poorer acuity outcome in the untreated group of hyperopes compared to controls (p < 0.0001) and to the children who were compliant in spectacle wear (p < 0.001) or who were prescribed spectacles (p < 0.05). Children who were significantly hyperopic at eight months were also more likely to be strabismic by 5.5 years compared to the emmetropic control group (p < 0.001). However, the present study did not find a significant difference in the incidence of strabismus between corrected and uncorrected hyperopic infants. Children who were not refractively corrected for significant hyperopia were four times more likely to have poor acuity at 5.5 years than infants who wore their hyperopic correction, supporting the findings of the First Cambridge Population Infant Vision Screening Programme.

Developmental trajectories for spatial frames of reference in Williams syndrome

Williams syndrome (WS) is a genetic disorder associated with severe visuocognitive impairment. Individuals with WS also report difficulties with everyday wayfinding. To study the development of body-, environment-, and object-based spatial frames of reference in WS, we tested 45 children and adults with WS on a search task in which the participant and a spatial array are moved with respect to each other. Although individuals with WS showed a marked delay, like young controls they demonstrated independent, additive use of body- and environment-based frames of reference. Crucially, object-based (intrinsic) representations based on local landmarks within the array were only marginally used even by adults with WS, whereas in typical development these emerge at 5 years. Deficits in landmark use are consistent with wayfinding difficulties in WS, and may also contribute to problems with basic localization, since in typical development landmark-based representations supplement those based on the body and on self-motion. Difficulties with inhibition or mental rotation may be further components in the impaired ability to use the correct reference frame in WS.

A Prospective Longitudinal Study of Retinal Structure and Function in Achromatopsia

PURPOSE To longitudinally characterize retinal structure and function in achromatopsia (ACHM) in preparation for clinical gene therapy trials. METHODS Thirty-eight molecularly confirmed ACHM subjects underwent serial assessments, including spectral domain optical coherence tomography (SD-OCT), microperimetry, and fundus autofluorescence (FAF). Foveal structure on SD-OCT was graded and compared for evidence of progression, along with serial measurements of foveal total retinal thickness (FTRT) and outer nuclear layer (ONL) thickness. Fundus autofluorescence patterns were characterized and compared over time. RESULTS Mean follow-up was 19.5 months (age range at baseline, 6-52 years). Only 2 (5%) of 37 subjects demonstrated change in serial foveal SD-OCT scans. There was no statistically significant change over time in FTRT (P = 0.83), ONL thickness (P = 0.27), hyporeflective zone diameter (P = 0.42), visual acuity (P = 0.89), contrast sensitivity (P = 0.22), mean retinal sensitivity (P = 0.84), and fixation stability (P = 0.58). Three distinct FAF patterns were observed (n = 30): central increased FAF (n = 4), normal FAF (n = 11), and well-demarcated reduced FAF (n = 15); with the latter group displaying a slow increase in the area of reduced FAF of 0.03 mm(2) over 19.3 months (P = 0.002). CONCLUSIONS Previously published cross-sectional studies have described conflicting findings with respect to the age-dependency of progression. This study, which constitutes the largest and longest prospective longitudinal study of ACHM to date, suggests that although ACHM may be progressive, any such progression is slow and subtle in most patients, and does not correlate with age or genotype. We also describe the first serial assessment of FAF, which is highly variable between individuals, even of similar age and genotype.

Weighted cue integration in the rodent head direction system

How the brain combines information from different sensory modalities and of differing reliability is an important and still-unanswered question. Using the head direction (HD) system as a model, we explored the resolution of conflicts between landmarks and background cues. Sensory cue integration models predict averaging of the two cues, whereas attractor models predict capture of the signal by the dominant cue. We found that a visual landmark mostly captured the HD signal at low conflicts: however, there was an increasing propensity for the cells to integrate the cues thereafter. A large conflict presented to naive rats resulted in greater visual cue capture (less integration) than in experienced rats, revealing an effect of experience. We propose that weighted cue integration in HD cells arises from dynamic plasticity of the feed-forward inputs to the network, causing within-trial spatial redistribution of the visual inputs onto the ring. This suggests that an attractor network can implement decision processes about cue reliability using simple architecture and learning rules, thus providing a potential neural substrate for weighted cue integration.

Refractive errors in infancy predict reduced performance on the Movement Assessment Battery for Children at 31/2 and 51/2 years

We have previously reported that significant hyperopia at 9 months predicts mild deficits on visuocognitive and visuomotor measures between 2 years and 5 years 6 months. Here we compare the motor skills of children who had been hyperopic in infancy (hyperopic group) with those who had been emmetropic (control group), using the Movement Assessment Battery for Children (Movement ABC). Children were tested at 3 years 6 months (hyperopic group: 47 males, 63 females, mean age 3y 7mo, SD 1.6mo; control group: 61 males, 70 females, mean age 3y 7mo, SD 1.2mo) and at 5 years 6 months (hyperopic group: 43 males, 56 females, mean age 5y 4mo, SD 1.7mo; control group: 51 males, 62 females, mean age 5y 3mo, SD 1.6mo). The hyperopic group performed significantly worse at both ages, overall and on at least one test from each category of motor skill (manual dexterity, balance, and ball skills). Distributions of scores showed that these differences were not due to poor performance by a minority but to a widespread mild deficit in the hyperopic group. This study also provides the first normative data on the Movement ABC for children below 4 years of age, and shows that it provides a useful measure of motor development at this young age.