July 2019
Volume 60, Issue 9
Free
ARVO Annual Meeting Abstract  |   July 2019
Comparative gene expression in proliferating versus differentiating human meibomian gland epithelial cells
Author Affiliations & Notes
  • David A Sullivan
    Schepens Eye Res Inst/Harvard Med School, Boston, Massachusetts, United States
  • Wendy R Kam
    Schepens Eye Res Inst/Harvard Med School, Boston, Massachusetts, United States
  • Yang Liu
    Schepens Eye Res Inst/Harvard Med School, Boston, Massachusetts, United States
  • Juan Ding
    Ophthalmology & Visual Sciences, UMass Memorial Medical Center, Worcester, Massachusetts, United States
  • Footnotes
    Commercial Relationships   David Sullivan, None; Wendy Kam, None; Yang Liu, None; Juan Ding, None
  • Footnotes
    Support  This research was supported by NIH grant EY05612, the Margaret S. Sinon Scholar in Ocular Surface Research fund and the and the AFER/Vistakon Dry Eye Fellowship
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 4191. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      David A Sullivan, Wendy R Kam, Yang Liu, Juan Ding; Comparative gene expression in proliferating versus differentiating human meibomian gland epithelial cells. Invest. Ophthalmol. Vis. Sci. 2019;60(9):4191.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : We recently discovered that Lrig1 and DNase2 serve as biomarkers for progenitor and differentiated cell populations, respectively, in the human meibomian gland (HMG). We also discovered that these biomarkers are plastic, and can be alternately induced or suppressed by changing the proliferation and differentiation status of immortalized (I) HMG epithelial cells (ECs). These changes are mirrored by corresponding shifts in cellular morphology in vitro. We hypothesize that this plasticity in biomarker profile and morphological appearance is paralleled by very significant alterations in cellular gene expression. To begin to test this hypothesis, we compared the gene expression patterns of proliferating versus differentiating IHMGECs.

Methods : IHMGECs were cultured for four days in either proliferating (keratinocyte serum free medium supplemented with epidermal growth factor and bovine pituitary extract] or differentiating [DMEM/F12 plus 10% fetal bovine serum] media. After four days of culture, cells were processed for the analysis of gene expression by using Illumina HumanHT-12 v.4 Expression BeadChips. Background-subtracted, cubic spline-normalized and non-log-transformed data were then evaluated with bioinformatic software.

Results : Our study identified significant differences in the expression of more than 9,200 genes in proliferating (4,918 genes upregulated) versus differentiating (4,364 genes upregulated) IHMGECs. Proliferation was associated with extraordinary increases in the expression of specific genes (e.g. cathelicidin antimicrobial peptide; 859,100-fold upregulation) and numerous ontologies (e.g. 152 biological process [bp] ontologies with ≥ 100 genes were upregulated), with the top 10 ontologies related to cell cycle (z scores > 13.9). Differentiation also led to dramatic increases in specific gene expressions (e.g. small proline-rich protein 3; 3,774,864-fold upregulation) and many ontologies (e.g. 134 bp ontologies with ≥ 100 genes were upregulated), with the protein and vesicle transport and lysosome ontologies having high z scores.

Conclusions : Our findings demonstrate that gene expression in proliferating versus differentiating IHMGECs is quite different. Our next objective is to determine whether these genetic differences exhibit plasticity like the biomarker proteins and cellular morphology of IHMGECs.

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

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×