Funduse sinine ja lähi-infrapuna autofluorestsentsuuring autosoom-retsessiivse Stardgardti tõve, koroidereemia, PROM1-maakuli düstroofia ja okulaarse albinismi patsientidel

Väitekirja elektrooniline versioon ei sisalda publikatsiooneFunduse sinine ja lähi-infrapuna autofluorestsentsuuring autosoom-retsessiivse Stardgardti tõve, koroidereemia, PROM1-maakuli düstroofia ja okulaarse albinismi patsientidel Pärilikud võrkkestahaigused on juhtivaks nägemiskaotuse põhjuseks tööealise elanikkonna seas arenenud riikides. Tegemist on kliiniliselt ja geneetiliselt väga heterogeense haiguste grupiga, mistõttu diagnostika ja haiguse patogeneesi uurimine on olnud vaevarikas. Võrkkesta piltdiagnostika on oluline mitte-invasiivne meetod haiguste diagnoosimiseks ja uurimiseks. Konfokaalne skanneeriv laseroftalmoskoop valgustab võrkkesta erineva lainepikkusega laserkiirega ning salvestab tagasikiirgavat valgust luues silmapõhjast pildi. Funduse autofluorestsents (AF) uuringul kasutatakse ära silmapõhja enda naturaalseid fluorofoore. Lipofustsiini ergastamiseks kasutatakse sinise spektri laserkiirt (sinine AF) ja melaniini jaoks lähipuna laserkiirt (lähipuna AF). Nende fluorofooride jaotus ja kogus silmapõhjas muutub erinevate haigusprotsesside mõjul ning need muutused on tuvastatavad AF uuringul. Antud doktoritöös uurisime sinise ja lähipuna AF uuringu pilte autosoom-retsesiivse Stargardti tõve (STGD1), koroidereemia, PROM1-maakuli düstroofia ning okulaarse albinismi patsientidel. Töö eesmärgiks oli paremini mõista sinise ja lähipuna AF signaali allikaid erinevate haigusseisundite korral, kus võrkkesta fluorofooride jaotus ning kogused on muutunud. Lisaks kvalitatiivsele piltide hindamisele kasutamise kvantitatiivset AF signaali tugevuse mõõtmist hindamaks lipofustsiini ja melaniini taset. Uurimustöös näitasime, et melaniin on lähipuna AF signaali peamiseks allikaks. Lisaks näitasime, et melanin võib kaudselt moduleerida lipofustsiinist tuleneva sinise AF signaali, sest okulaarse albinismi kandjate hüpopigmenteeritud võrkkesta alade sinise AF signal oli tavapärasest kõrgem. AF signaali tugevuse mõõtmisel leidsime, et lipofustsiini kuhjumine võrkkestas põhjustab lisaks sinise AF signaali tõusule ka lähipuna AF signaali tõusu STGD1 patsientidel. Kvantitatiivsel analüüsil näitasime ka, et PROM1-maakuli düstroofia patsientide sinise AF signaal oli võrreldav terve silmapõhja signaali tugevusega, eristades seda fenotüübiliselt sarnasest STGD1 haigusest ning viidates ka sellele, et lipofustsiini üleliigne kuhjumine ei ole antud haigusele omane mehhanism. Koroidereemia ja STGD1 haigete uurimisel leidsime, et pigmentepiteeli rakkude kärbumine on nähtav AF signaali hääbumisena, samas lähipuna AF uuringaitab tuvastada varasemaid muutusi kui sinine AF uuring. Lipofustsiin ja melanin on mõlemad olulised võrkkesta rakkude seisundi biomarkerid, mida on võimalik mitte-invasiivsel moel AF uuringu abil analüüsida ning hinnata haiguse progressiooni.Inherited retinal diseases are the leading cause of visual impairment among the working age-group in the developed countries. Because of genetic and phenotypical heterogeneity, diagnosis and understanding pathogenesis of inherited retinal disease has been challenging. Retinal imaging studies which are noninvasive, are an invaluable source of information. Fundus autofluorescence (FAF) utilizes natural fluorophores to create an image of the retina. Lipofuscin is the primary source for short-wavelength autofluorescence (SW-AF) and melanin for near-infrared autofluorescence (NIR-AF). The amount and distribution of these fluorophores changes in the different disease processes and is detectable in FAF images. In this study we analyzed SW-AF and NIR-AF images in cases of genetically confirmed recessive Stargardt disease (STGD1), choroideremia, PROM1-macular disease and ocular albinism. The aim was to qualitatively describe FAF in conditions with varying levels of lipofuscin or melanin as well as to quantify FAF signal intensities. We also aimed at finding new clinical implications for autofluorescence imaging in evaluating inherited retinal disease. We confirmed that melanin is the major source of NIR-AF signal by analyzing ocular albinism carriers and mice models with varying fundus pigmentation, but we also found that presence of melanin can modulate SW-AF signal strength. As a novel finding we confirmed that lipofuscin contributes to NIR-AF signal intensity in cases with excessive bisretinoid lipofuscin levels like seen in STGD1. The analysis of choroideremia and STGD1 patients showed that retinal pigment epithelium atrophy causes loss of signal in both SW-AF and NIR-AF, but NIR-AF could be more sensitive in detecting early cell degeneration. Quantifying the autofluorescence signal intensity helps to further understand disease processes as it is an indirect measure for levels of retinal fluorophores. We showed PROM1-macular dystrophy does not present with elevated levels of SW-AF indicating that excessive lipofuscin accumulation is likely not part of its disease mechanism. That knowledge is valuable in differentiating it from phenotypically similar STGD1 or when developing therapeutic approaches. Lipofuscin and melanin are both valuable retinal biomarkers for evaluating retinal health by using non-invasive autofluorescence imaging.https://www.ester.ee/record=b555738

Intraretinal Correlates of Reticular Pseudodrusen Revealed by Autofluorescence and En Face OCT.

Purpose We sought to determine whether information revealed from the reflectance, autofluorescence, and absorption properties of RPE cells situated posterior to reticular pseudodrusen (RPD) could provide insight into the origins and structure of RPD. Methods RPD were studied qualitatively by near-infrared fundus autofluorescence (NIR-AF), short-wavelength fundus autofluorescence (SW-AF), and infrared reflectance (IR-R) images, and the presentation was compared to horizontal and en face spectral domain optical coherence tomographic (SD-OCT) images. Images were acquired from 23 patients (39 eyes) diagnosed with RPD (mean age 80.7 ± 7.1 [SD]; 16 female; 4 Hispanics, 19 non-Hispanic whites). Results In SW-AF, NIR-AF, and IR-R images, fundus RPD were recognized as interlacing networks of small scale variations in IR-R and fluorescence (SW-AF, NIR-AF) intensities. Darkened foci of RPD colocalized in SW-AF and NIR-AF images, and in SD-OCT images corresponded to disturbances of the interdigitation (IZ) and ellipsoid (EZ) zones and to more pronounced hyperreflective lesions traversing photoreceptor-attributable bands in SD-OCT images. Qualitative assessment of the outer nuclear layer (ONL) revealed thinning as RPD extended radially from the outer to inner retina. In en face OCT, hyperreflective areas in the EZ band correlated topographically with hyporeflective foci at the level of the RPE. Conclusions The hyperreflective lesions corresponding to RPD in SD-OCT scans are likely indicative of degenerating photoreceptor cells. The darkened foci at positions of RPD in NIR-AF and en face OCT images indicate changes in the RPE monolayer with the reduced NIR-AF and en face OCT signal suggesting a reduction in melanin that could be accounted for by RPE thinning

Hyperautofluorescent Dots are Characteristic in Ceramide Kinase Like-associated Retinal Degeneration.

There is a lack of studies which seek to discern disease expression in patients with mutations that alter retinal ceramide metabolism, specifically in the ceramide kinase like (CERKL) gene. This cross-sectional case series reports a novel phenotypic manifestation of CERKL-associated retinopathy. Four unrelated patients with homozygous CERKL mutations underwent a complete ocular exam, spectral-domain optical coherence tomography, short-wavelength fundus autofluorescence (SW-AF), quantitative autofluorescence (qAF), and full-field electroretinogram (ffERG). Decreased visual acuity and early-onset maculopathy were present in all patients. All four patients had extensive hyperautofluorescent foci surrounding an area of central atrophy on SW-AF imaging, which has not been previously characterized. An abnormal spatial distribution of qAF signal was seen in one patient, and abnormally elevated qA

Patterns and Intensities of Near-Infrared and Short-Wavelength Fundus Autofluorescence in Choroideremia Probands and Carriers

PURPOSE. To ascertain cellular constituents within islands of preserved retina in choroideremia (CHM) by multimodal imaging. METHODS. CHM probands (16) and female carriers (9) of CHM were studied. Near-infrared autofluorescence (NIR-AF; 787-nm excitation; emission, > 830 nm), short-wavelength autofluorescence (SW-AF; 488-nm excitation, 500-to 680-nm emission), and spectral-domain optical coherence tomography (SD-OCT) images were acquired with a confocal scanning laser ophthalmoscope. SW-AF intensities were measured by quantitative fundus autofluorescence (qAF), and NIR-AF intensity profiles were analyzed. Retinal thicknesses and visual acuity were measured. RESULTS. In 19 of 31 eyes of affected males, islands of preserved NIR-AF signal were also visible as fluorescence signal in SW-AF images. Notable in 12 eyes were areas of speckled SW-AF that was hypoautofluorescent in the NIR-AF image. Islands of preserved NIR-AF and SW-AF signal were often associated with the presence of visible but thinned outer nuclear layer and discontinuous interdigitation zone, ellipsoid zone, and external limiting membrane. NIR-AF profiles revealed that even in areas of preserved retina, the NIR-AF signal from retinal pigment epithelium (RPE) melanin is greatly reduced. qAF was reduced overall. The fundus of carriers was characterized by a mosaicism in which patches of reduced NIR-AF colocalized with reduced SW-AF. CONCLUSIONS. In CHM-affected males, the presence of RPE was indicated by an NIR-AF signal and the absence of hypertransmission of OCT signal into the choroid. RPE preservation was associated with better visual acuity. In carriers, patches of reduced SW-AF colocalized with decreased NIR-AF and qAF was severely reduced.Peer reviewe

Insights Into PROM1-Macular Disease Using Multimodal Imaging

PURPOSE. To describe the features of genetically confirmed PROM1-macular dystrophy in multimodal images. METHODS. Thirty-six (36) eyes of 18 patients (5-66 years; mean age, 42.4 years) were prospectively studied by clinical examination and multimodal imaging. Short-wavelength autofluorescence (SW-AF) and quantitative fundus autofluorescence (qAF) images were acquired with a scanning laser ophthalmoscope (HRA+OCT, Heidelberg Engineering) modified by insertion of an internal autofluorescent reference. Further clinical testing included near-infrared autofluorescence (NIR-AF; HRA2, Heidelberg Engineering) with semiquantitative analysis, spectral domain-optical coherence tomography (HRA+OCT) and full-field electroretinography. All patients were genetically confirmed by exome sequencing. RESULTS. All 18 patients presented with varying degrees of maculopathy. One family with individuals affected across two generations exhibited granular fleck-like deposits across the posterior pole. Areas of granular deposition in SW-AF and NIR-AF corresponded to intermittent loss of the ellipsoid zone, whereas discrete regions of hypoautofluorescence corresponded with a loss of outer retinal layers in spectral-domain optical coherence tomography scans. For 18 of the 20 eyes, qAF levels within the macula were within the 95% confidence intervals of healthy age-matched individuals; nor was the mean NIR-AF signal increased relative to healthy eyes. CONCLUSIONS. Although PROM1-macular dystrophy (Stargardt disease 4) can exhibit phenotypic overlap with recessive Stargardt disease, significantly increased SW-AF levels were not detected. As such, elevated bisretinoid lipofuscin may not be a feature of the pathophysiology of PROM1 disease. The qAF approach could serve as a method of early differential diagnosis and may help to identify appropriate disease targets as therapeutics become available to treat inherited retinal disease.Peer reviewe