CRB1-associated Retinal Dystrophies: Genetics, Clinical Characteristics and Natural History

PURPOSE: To analyse the clinical characteristics, natural history, and genetics of CRB1-associated retinal dystrophies. DESIGN: Multicenter international retrospective cohort study. METHODS: Review of clinical notes, ophthalmic images, and genetic testing results of 104 patients (91 probands) with disease-causing CRB1 variants. Macular optical coherence tomography (OCT) parameters, visual function, fundus characteristics, and associations between variables were our main outcome measures. RESULTS: The mean age of the cohort at the first visit was 19.8 ± 16.1 (median 15) years of age, with a mean follow-up of 9.6 ± 10 years. Based on history, imaging, and clinical examination, 26 individuals were diagnosed with retinitis pigmentosa (RP, 26%), 54 with early-onset severe retinal dystrophy/Leber Congenital Amaurosis (EOSRD/LCA, 51%), and 24 with macular dystrophy (MD, 23%). Severe visual impairment was most frequent after 40 years of age for patients with RP and after 20 years of age for EOSRD/LCA. Longitudinal analysis revealed a significant difference between baseline and follow up best corrected visual acuity in the three sub-cohorts. Macular thickness decreased in most patients with EOSRD/LCA and MD, whereas the majority of patients with RP had increased perifoveal thickness. CONCLUSIONS: A subset of individuals with CRB1 variants present with mild, adult-onset RP. EOSRD/LCA phenotype was significantly associated with null variants, and 167_169 deletion was exclusively present in the MD cohort. The poor OCT lamination may have a degenerative component, as well as being congenital. Disease symmetry and reasonable window for intervention highlight CRB1 retinal dystrophies as a promising target for trials of novel therapeutics

Resolving the dark matter of ABCA4 for 1054 Stargardt disease probands through integrated genomics and transcriptomics

Purpose: Missing heritability in human diseases represents a major challenge, and this is particularly true for ABCA4-associated Stargardt disease (STGD1). We aimed to elucidate the genomic and transcriptomic variation in 1054 unsolved STGD and STGD-like probands. Methods: Sequencing of the complete 128-kb ABCA4 gene was performed using single-molecule molecular inversion probes (smMIPs), based on a semiautomated and cost-effective method. Structural variants (SVs) were identified using relative read coverage analyses and putative splice defects were studied using in vitro assays. Results: In 448 biallelic probands 14 known and 13 novel deep-intronic variants were found, resulting in pseudoexon (PE) insertions or exon elongations in 105 alleles. Intriguingly, intron 13 variants c.1938-621G>A and c.1938-514G>A resulted in dual PE insertions consisting of the same upstream, but different downstream PEs. The intron 44 variant c.6148-84A>T resulted in two PE insertions and flanking exon deletions. Eleven distinct large deletions were found, two of which contained small inverted segments. Uniparental isodisomy of chromosome 1 was identified in one proband. Conclusion: Deep sequencing of ABCA4 and midigene-based splice assays allowed the identification of SVs and causal deep-intronic variants in 25% of biallelic STGD1 cases, which represents a model study that can be applied to other inherited diseases

Resolving the dark matter of ABCA4 for 1054 Stargardt disease probands through integrated genomics and transcriptomics

Purpose: Missing heritability in human diseases represents a major challenge, and this is particularly true for ABCA4-associated Stargardt disease (STGD1). We aimed to elucidate the genomic and transcriptomic variation in 1054 unsolved STGD and STGD-like probands. Methods: Sequencing of the complete 128-kb ABCA4 gene was performed using single-molecule molecular inversion probes (smMIPs), based on a semiautomated and cost-effective method. Structural variants (SVs) were identified using relative read coverage analyses and putative splice defects were studied using in vitro assays. Results: In 448 biallelic probands 14 known and 13 novel deep-intronic variants were found, resulting in pseudoexon (PE) insertions or exon elongations in 105 alleles. Intriguingly, intron 13 variants c.1938-621G>A and c.1938-514G>A resulted in dual PE insertions consisting of the same upstream, but different downstream PEs. The intron 44 variant c.6148-84A>T resulted in two PE insertions and flanking exon deletions. Eleven distinct large deletions were found, two of which contained small inverted segments. Uniparental isodisomy of chromosome 1 was identified in one proband. Conclusion: Deep sequencing of ABCA4 and midigene-based splice assays allowed the identification of SVs and causal deep-intronic variants in 25% of biallelic STGD1 cases, which represents a model study that can be applied to other inherited diseases.Fil: Khan, Mubeen. Radboud University Nijmegen Medical Centre; Países BajosFil: Cornelis, Stéphanie S.. Radboud University Nijmegen Medical Centre; Países BajosFil: Del Pozo Valero, Marta. Hospital Universitario Fundación Jiménez Díaz; España. Radboud University Nijmegen Medical Centre; Países BajosFil: Whelan, Laura. Trinity College; Estados UnidosFil: Runhart, Esmee H.. Radboud University Nijmegen Medical Centre; Países BajosFil: Mishra, Ketan. Radboud University Nijmegen Medical Centre; Países BajosFil: Bults, Femke. Radboud University Nijmegen Medical Centre; Países BajosFil: AlSwaiti, Yahya. St John of Jerusalem Eye Hospital Group; Palestina (ANP)Fil: AlTalbishi, Alaa. St John of Jerusalem Eye Hospital Group; Palestina (ANP)Fil: De Baere, Elfride. University of Ghent; BélgicaFil: Banfi, Sandro. Seconda Universita Degli Studi Di Napoli; ItaliaFil: Banin, Eyal. The Hebrew University of Jerusalem; IsraelFil: Bauwens, Miriam. University of Ghent; BélgicaFil: Ben Yosef, Tamar. The Ruth And Bruce Rappaport Faculty Of Medicine; IsraelFil: Boon, Camiel J. F.. Leiden University. Leiden University Medical Center; Países BajosFil: van den Born, L. Ingeborgh. Rotterdam Ophthalmic Institute; Países BajosFil: Defoort, Sabine. Universite Lille; FranciaFil: Devos, Aurore. Universite Lille; FranciaFil: Dockery, Adrian. Trinity College; Estados UnidosFil: Dudakova, Lubica. Charles University and General University Hospital; República ChecaFil: Fakin, Ana. Charles University and General University Hospital; República ChecaFil: Farrar, G. Jane. Trinity College; Estados UnidosFil: Ferraz Sallum, Juliana Maria. Universidade Federal de Sao Paulo; BrasilFil: Fujinami, Kaoru. UCL Institute of Ophthalmology; Reino UnidoFil: Gilissen, Christian. Radboud University Nijmegen Medical Centre; Países BajosFil: Glavac, Damjan. University of Ljubljana; EsloveniaFil: Gorin, Michael B.. University of California at Los Angeles. School of Medicine; Estados UnidosFil: Greenberg, Jacquie. University of Cape Town; SudáfricaFil: Hayashi, Takaaki. The Jikei University School of Medicine; JapónFil: Hettinga, Ymkje M.. Bartiméus Diagnostic Center for Complex Visual Disorders; Países BajosFil: Hoischen, Alexander. Radboud University Nijmegen Medical Centre; Países BajosFil: Hoyng, Carel B.. Radboud University Nijmegen Medical Centre; Países BajosFil: Hufendiek, Karsten. University Eye Hospital Hannover Medical School; AlemaniaFil: Jägle, Herbert. University Regensburg; AlemaniaFil: Kamakari, Smaragda. OMMA Ophthalmological Institute of Athens; GreciaFil: Karali, Marianthi. Seconda Universita Degli Studi Di Napoli; ItaliaFil: Kellner, Ulrich. No especifíca;Fil: Klaver, Caroline C. W.. Radboud University Nijmegen Medical Centre; Países BajosFil: Kousal, Bohdan. Charles University and General University Hospital; República ChecaFil: Lamey, Tina M.. University of Western Australia; AustraliaFil: MacDonald, Ian M.. University of Alberta; CanadáFil: Matynia, Anna. University of California at Los Angeles. School of Medicine; Estados UnidosFil: McLaren, Terri L.. University of Western Australia; AustraliaFil: Mena, Marcela D.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Meunier, Isabelle. Université Montpellier II; FranciaFil: Miller, Rianne. Radboud University Nijmegen Medical Centre; Países BajosFil: Newman, Hadas. Universitat Tel Aviv; IsraelFil: Ntozini, Buhle. University of Cape Town; SudáfricaFil: Oldak, Monika. No especifíca;Fil: Pieterse, Marc. Radboud University Nijmegen Medical Centre; Países BajosFil: Podhajcer, Osvaldo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Puech, Bernard. Universite Lille; FranciaFil: Ramesar, Raj. University of Cape Town; SudáfricaFil: Rüther, Klaus. No especifíca;Fil: Salameh, Manar. No especifíca;Fil: Salles, Mariana Vallim. Universidade de Sao Paulo; BrasilFil: Sharon, Dror. The Hebrew University of Jerusalem; IsraelFil: Simonelli, Francesca. Seconda Universita Degli Studi Di Napoli; ItaliaFil: Spital, Georg. No especifíca;Fil: Steehouwer, Marloes. Radboud University Nijmegen Medical Centre; Países BajosFil: Szaflik, Jacek P.. No especifíca;Fil: Thompson, Jennifer A.. No especifíca;Fil: Thuillier, Caroline. Universite Lille; FranciaFil: Tracewska, Anna M.. No especifíca;Fil: van Zweeden, Martine. Radboud University Nijmegen Medical Centre; Países BajosFil: Vincent, Andrea L.. University of Auckland; Nueva ZelandaFil: Zanlonghi, Xavier. No especifíca;Fil: Liskova, Petra. Charles University and General University Hospital; República ChecaFil: Stöhr, Heidi. Universitat Regensburg; AlemaniaFil: De Roach, John N.. University of Western Australia; AustraliaFil: Ayuso, Carmen. Hospital Universitario Fundación Jiménez Díaz; EspañaFil: Roberts, Lisa. University of Cape Town; SudáfricaFil: Weber, Bernhard H. F.. Universitat Regensburg; AlemaniaFil: Dhaenens, Claire Marie. Universite Lille; FranciaFil: Cremers, Frans P. M.. Radboud University Nijmegen Medical Centre; Países Bajo