June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Genomic and functional approaches in identification of retinal disease genes and new insights to centrosome and cilia biology in the retina
Author Affiliations & Notes
  • Amin Sabri
    Eye Genetics Research Unit, Children's Medical Research Institute, University of Sydney, Sydney, New South Wales, Australia
  • Benjamin Nash
    Eye Genetics Research Unit, Children's Medical Research Institute, University of Sydney, Sydney, New South Wales, Australia
    Discipline of Genetic Medicine, Children’s Hospital at Westmead Clinical School, Sydney Medical School, University of Sydney, Sydney, New South Wales, Australia
  • Anson Cheng
    Eye Genetics Research Unit, Children's Medical Research Institute, University of Sydney, Sydney, New South Wales, Australia
    Save Sight Institute, University of Sydney, Sydney, New South Wales, Australia
  • Rebecca Greenlees
    Eye Genetics Research Unit, Children's Medical Research Institute, University of Sydney, Sydney, New South Wales, Australia
  • Bruce Bennetts
    Discipline of Genetic Medicine, Children’s Hospital at Westmead Clinical School, Sydney Medical School, University of Sydney, Sydney, New South Wales, Australia
    Sydney Genome Diagnostics, Western Sydney Genetics Program, Sydney Children’s Hospital Network, Sydney, New South Wales, Australia
  • John R Grigg
    Eye Genetics Research Unit, Children's Medical Research Institute, University of Sydney, Sydney, New South Wales, Australia
    Save Sight Institute, University of Sydney, Sydney, New South Wales, Australia
  • Robyn Jamieson
    Eye Genetics Research Unit, Children's Medical Research Institute, University of Sydney, Sydney, New South Wales, Australia
    Discipline of Genetic Medicine, Children’s Hospital at Westmead Clinical School, Sydney Medical School, University of Sydney, Sydney, New South Wales, Australia
  • Footnotes
    Commercial Relationships   Amin Sabri, None; Benjamin Nash, None; Anson Cheng, None; Rebecca Greenlees, None; Bruce Bennetts, None; John Grigg, None; Robyn Jamieson, None
  • Footnotes
    Support  NHMRC Australia
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 4513. doi:
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      Amin Sabri, Benjamin Nash, Anson Cheng, Rebecca Greenlees, Bruce Bennetts, John R Grigg, Robyn Jamieson; Genomic and functional approaches in identification of retinal disease genes and new insights to centrosome and cilia biology in the retina. Invest. Ophthalmol. Vis. Sci. 2017;58(8):4513.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Retinal dystrophy (RD) is a significant cause of vision loss. Many of the underlying disease genes and their functions are not known. We are using genomic approaches to identify variants in known disease genes, followed by strategies for novel disease gene identification where required. Our purpose is to provide molecular diagnoses for affected individuals, as well new insights to retinal and cell biology.

Methods : We applied Trusight One Clinical Exome capture (Illumina Inc CA USA), followed by sequencing on the Illumina HiSeq2500. Further genomic investigation was undertaken for known and novel disease gene identification. Variants in known and candidate RD genes were filtered and prioritized in 50 sporadic and familial RD patients. Expression studies were performed in mouse tissues and human cell lines. Functional studies were performed in mutant and control human fibroblasts, and transfected ARPE-19 and HeLa cells.

Results : Pathogenic and likely pathogenic variants were detected in over 70% of our RD cohort. Clear-cut causative variants were identified in the majority of cases, and results also included unexpected findings in CHM, and combinations with variants in more than one RD gene, including EYS and RP1, requiring detailed phenotypic and segregation analysis. We identified a novel retinal disease gene which our analysis implicates in centrosome and cilia biology. Expression analyses showed presence of the encoded protein of this gene in the retinal pigmented epithelium and inner segments of the photoreceptors. Cell-based assays revealed its localization in centrosomes during interphase, and the basal body of the primary cilium in non-dividing cells. Immunocytochemistry on skin fibroblasts revealed a higher percentage of cells with abnormal numbers of centrosomes in affected individuals. In HeLa cells transfected with the mutant construct, there were a higher number of multinucleated cells, further suggesting abnormality of centrosome and cell cycle biology.

Conclusions : Genomic investigation is valuable in RD and this study highlights benefits of genetic and phenotypic multidisciplinary review for accurate diagnosis. Our genomic and functional work has also led to new knowledge with implications for regulation of centrosome biology and impact on the cell cycle and ciliary function in the retina.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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