June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Differential gene expression by human lens epithelial cells in post vitrectomy cataract vs. steroid induced cataract revealed by RNA sequencing
Author Affiliations & Notes
  • Michael Dong
    Central Clinical School, Monash University Faculty of Medicine Nursing and Health Sciences, Clayton, Victoria, Australia
    Department of Ophthalmology, Alfred Health, Melbourne, Victoria, Australia
  • Jaynish Shah
    Myeloma Research Group, Australian Centre for Blood Diseases, The Alfred Hospital/Monash University, Melbourne, Victoria, Australia
  • Lyndell L Lim
    Centre for Eye Research Australia, University of Melbourne, East Melbourne, Victoria, Australia
    Royal Victorian Eye and Ear Hospital, East Melbourne, Victoria, Australia
  • Tiffany Khong
    Myeloma Research Group, Australian Centre for Blood Diseases, The Alfred Hospital/Monash University, Melbourne, Victoria, Australia
  • Andrew Spencer
    Myeloma Research Group, Australian Centre for Blood Diseases, The Alfred Hospital/Monash University, Melbourne, Victoria, Australia
  • Anthony John Hall
    Central Clinical School, Monash University Faculty of Medicine Nursing and Health Sciences, Clayton, Victoria, Australia
    Department of Ophthalmology, Alfred Health, Melbourne, Victoria, Australia
  • Footnotes
    Commercial Relationships   Michael Dong None; Jaynish Shah None; Lyndell Lim None; Tiffany Khong None; Andrew Spencer None; Anthony Hall None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 1145. doi:
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    • Get Citation

      Michael Dong, Jaynish Shah, Lyndell L Lim, Tiffany Khong, Andrew Spencer, Anthony John Hall; Differential gene expression by human lens epithelial cells in post vitrectomy cataract vs. steroid induced cataract revealed by RNA sequencing. Invest. Ophthalmol. Vis. Sci. 2022;63(7):1145.

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

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Abstract

Purpose : Vitrectomy and corticosteroid-use are known to accelerate cataract development, however, the causative mechanisms are largely unknown. We conducted RNA-sequencing of human lens epithelial cells in steroid-induced and post-vitrectomy cataract to verify and compare gene expression profiles.

Methods : Human anterior lens capsule specimens from post-vitrectomy (n = 8), steroid-induced subcapsular (n = 12) and age-related (n = 2) cataracts were collected from patients following written informed consent from the Alfred Hospital Research and Ethics Committee. Capsulorhexis specimens were collected at the time of cataract surgery for analysis. RNA was isolated using RNeasy Micro Kit and RNA libraries were synthesized using Illumina Stranded Total RNA Prep Ligation with Ribo-Zero Plus followed by sequencing on a NovaSeq S4, 300 cycles at AGRF. Differentially expressed genes (DEGs) were identified and underwent bioinformatics analyses including principal component analysis (PCA).

Results : Approximately 55,000 genes were detected from RNA-seq. We focused on ~15,000 genes with moderate or high average expression of >10 transcripts per million reads sequenced. A dimensionality reduction algorithm known as Principal Component Analysis (PCA) was employed to study gross differences in gene expression profile. PCA clustered steroid induced subcapsular cataract and post-vitrectomy cataract separately (Figure 1), suggesting overall differences in gene expression between the two groups (Figure 2), while age-related cataract samples clustered close to post-vitrectomy samples. Preliminary results indicate a minimum of 28 identified DEGs, 27 were upregulated in post-vitrectomy compared to steroid-induced cataract including IL-33, P2RY6, CAMP, CP, C4B, CFI whilst GPD1 was downregulated.

Conclusions : Our findings suggest that post-vitrectomy cataract and steroid-induced subcapsular cataract have different RNA transcription profiles. Bioinformatics analyses suggest that lens epithelial cells from post-vitrectomy cataract have a similar transcriptome to lens epithelial cells from age-related cataract but the transcriptome from steroid-induced posterior subcapsular cataract lens epithelial cells was quantitatively and qualitatively different.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

 

Figure 1, PCA of normalized RNA-seq counts

Figure 1, PCA of normalized RNA-seq counts

 

Figure 2, DEGs in post-vitrectomy cataract vs steroid-induced posterior subcapsular cataract

Figure 2, DEGs in post-vitrectomy cataract vs steroid-induced posterior subcapsular cataract

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