Next‐generation genetic testing for retinitis pigmentosa

Abstract

Molecular diagnostics for patients with retinitis pigmentosa (RP) has been hampered by extreme genetic and clinical heterogeneity, with 52 causative genes known to date. Here, we developed a comprehensive next-generation sequencing (NGS) approach for the clinical molecular diagnostics of RP. All known inherited retinal disease genes (n = 111) were captured and simultaneously analyzed using NGS in 100 RP patients without a molecular diagnosis. A systematic data analysis pipeline was developed and validated to prioritize and predict the pathogenicity of all genetic variants identified in each patient, which enabled us to reduce the number of potential pathogenic variants from approximately 1,200 to zero to nine per patient. Subsequent segregation analysis and in silico predictions of pathogenicity resulted in a molecular diagnosis in 36 RP patients, comprising 27 recessive, six dominant, and three X-linked cases. Intriguingly, De novo mutations were present in at least three out of 28 isolated cases with causative mutations. This study demonstrates the enormous potential and clinical utility of NGS in molecular diagnosis of genetically heterogeneous diseases such as RP. De novo dominant mutations appear to play a significant role in patients with isolated RP, having major implications for genetic counselling.

Next-generation sequencing applied to a large French cone and cone-rod dystrophy cohort: mutation spectrum and new genotype-phenotype correlation

“…MLPA did not identify large deletions for these cases. In addition, a single published nonsense variant was identified in IMPG2 at a heterozygous state in one sporadic case (CIC00680, c.379G>A p.(R127*)) [16] and another novel single variant also heterozygous was found in CDHR1 (CIC07507, c.1863dupC p.(I622H*fs54)) in an ar case with consanguineous parents (Additional file 3). Sanger sequencing confirmed these two variants.…”

Autosomal recessive retinitis pigmentosa withRP1mutations is associated with myopia

“…The study was done under the authorisation of the Department of Ophthalmology of the Hospital of Montpellier # 11018S from the French Ministry of Health for biomedical research in the field of physiology, pathophysiology, epidemiology and genetics in ophthalmology. A total of 26 RP patients who were either homozygotes or compound heterozygotes for RP1 mutations were ascertained (table 1), including 6 originating from France and Morocco,15 20 11 from Spain,17 21 4 from Saudi Arabia,18 3 from Pakistan11and 2 from the Netherlands 16 19. Patients who had cataract surgery or aphakic eyes were excluded from the study.…”

Comprehensive molecular diagnosis of 179 Leber congenital amaurosis and juvenile retinitis pigmentosa patients by targeted next generation sequencing

“…With additional mutations added to the LCA APEX array, the estimated solving rate has been improved to about 50% 11. On the other hand, next generation sequencing (NGS) has been recently used for the molecular diagnosis of retinal diseases 17 18. Compared with the APEX chip, the NGS based approach is able to discover novel variants and genes.…”