July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Lamellar Cataract Transcriptome Analyses Suggest Developmental Delay in The Onset of the Differentiation Program in the Ocular Lens
Author Affiliations & Notes
  • Rajendra K Gangalum
    Ophthalmology, Stein Eye Institute, UCLA, Los Angeles, California, United States
  • Matthew Brooks
    National Eye Institute, N.I.H., Bethesda, Maryland, United States
  • Dongjae Kim
    Stein Eye Institute, UCLA, Los Angeles, California, United States
  • Linn Gieser
    National Eye Institute, N.I.H., Bethesda, Maryland, United States
  • Anand Swaroop
    National Eye Institute, N.I.H., Bethesda, Maryland, United States
  • Suraj P Bhat
    Stein Eye Institute, UCLA, Los Angeles, California, United States
    Molecular Biology Institute and Brain Research Institute, UCLA, Los Angeles, California, United States
  • Footnotes
    Commercial Relationships   Rajendra Gangalum, None; Matthew Brooks, None; Dongjae Kim, None; Linn Gieser, None; Anand Swaroop, None; Suraj Bhat, None
  • Footnotes
    Support  RKG is supported by NIH grant (1R01EY024929) to SPB. SPB is a Research To Prevent Blindness Inc., Wasserman Merit Scholar. This work was also supported the Gerald Oppenheimer Family Foundation for the Prevention of Eye Disease Endowment Fund.
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 4298. doi:https://doi.org/
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      Rajendra K Gangalum, Matthew Brooks, Dongjae Kim, Linn Gieser, Anand Swaroop, Suraj P Bhat; Lamellar Cataract Transcriptome Analyses Suggest Developmental Delay in The Onset of the Differentiation Program in the Ocular Lens. Invest. Ophthalmol. Vis. Sci. 2019;60(9):4298. doi: https://doi.org/.

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

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Abstract

Purpose : Childhood lamellar cataracts are the most prevalent cataracts. These cataracts are associated with mutations in the heat shock transcription factor 4 (Hsf4). We have generated lamellar cataract mouse model by introducing R116H mutation. The cataract phenotype is subtle, with opacities confined to few lamellae in the lens nucleus. In this study, we investigate the transcriptome of the developing mutant lens to understand molecular progression that leads to cataractogenesis.

Methods : The lens development in the transgenic mutant (R116H) was followed by 2D-gel analysis of single lens proteins. We used SMARTer stranded total RNA-seq strategies (Takara Bio USA Inc) for transcriptomic analysis of developing ocular lens at post-natal day2, (D02), day10 (D10) and day25 (D25) stages. The data was processed (FastQC) and annotated (Ensembl v84) and analysed using edgeR(TMM).

Results : Comparison of protein coding transcripts in the WT and the mutant lens, does not show any significant quantitative changes at D02 (20137-WT and 21751- R116H), D10 (21369-WT and 21543-R116H) or at D25 (21509-WT and 21724-R116H). However, we see alterations in the total expression patterns of transcripts unique to each temporal stage. For example, The number of unique transcripts in WT at D02, D10 and D25 are 79, 128 and 195 respectively. In the mutant, they are 44, 20 and 67 suggesting an inhibited program of differentiation. Unsupervised clustering indicates that crystallin gene expression increase appreciably from D02 to D10 and then dwindles by D25 in WT. In the mutant, the crystallin expression is poor both at D02 and D10, yet at D25 we see an increase in selective crystallin transcripts. This inference is also reinforced by the examination of the 2D gel patterns of these predominant proteins. Interestingly, transcript levels of Filensin (Bfsp1) (associated with early onset nuclear cataracts) and DNase 2B, that is associated with nuclear degeneration in terminal differentiation were decreased.

Conclusions : Based on the complete analysis of the gene expression profiles of the protein coding transcripts at three developmental stages in the WT and the mutant lens we see stalled developmental programs associated with cataractogenesis in the Lamellar cataract transgenic mouse paradigm.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

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