June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Differential miRNA Expression in Response to Mechanical Stress in Human Trabecular Meshwork Cells
Author Affiliations & Notes
  • Inas Helwa
    Cellular Biology and Anatomy, Augusta University, Augusta, Georgia, United States
  • Michelle Drewry
    Cellular Biology and Anatomy, Augusta University, Augusta, Georgia, United States
  • William M Johnson
    Ophthalmology, Duke University Medical Center, Durham, North Carolina, United States
  • W. Michael Dismuke
    Ophthalmology, Duke University Medical Center, Durham, North Carolina, United States
  • Inas F Aboobakar
    Ophthalmology, Duke University Medical Center, Durham, North Carolina, United States
  • Kristin Marie Perkumas
    Ophthalmology, Duke University Medical Center, Durham, North Carolina, United States
  • R. Rand Allingham
    Ophthalmology, Duke University Medical Center, Durham, North Carolina, United States
  • Michael A Hauser
    Ophthalmology, Duke University Medical Center, Durham, North Carolina, United States
    Medicine, Duke University Medical Center, Durham, North Carolina, United States
  • W. Daniel Stamer
    Ophthalmology, Duke University Medical Center, Durham, North Carolina, United States
  • Yutao Liu
    Cellular Biology and Anatomy, Augusta University, Augusta, Georgia, United States
  • Footnotes
    Commercial Relationships   Inas Helwa, None; Michelle Drewry, None; William Johnson, None; W. Michael Dismuke, None; Inas Aboobakar, None; Kristin Perkumas, None; R. Allingham, None; Michael Hauser, None; W. Stamer, None; Yutao Liu, None
  • Footnotes
    Support  Glaucoma Research Foundation, Bright Focus Foundation, Glaucoma Foundation, R01EY019696, R01EY02387
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 3481. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to Subscribers Only
      Sign In or Create an Account ×
    • Get Citation

      Inas Helwa, Michelle Drewry, William M Johnson, W. Michael Dismuke, Inas F Aboobakar, Kristin Marie Perkumas, R. Rand Allingham, Michael A Hauser, W. Daniel Stamer, Yutao Liu; Differential miRNA Expression in Response to Mechanical Stress in Human Trabecular Meshwork Cells. Invest. Ophthalmol. Vis. Sci. 2017;58(8):3481.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : miRNAs have been shown to play important roles in regulating trabecular meshwork (TM) function and aqueous outflow. We aimed to identify differentially expressed miRNAs in human TM (HTM) cells in response to long-term mechanical stress.

Methods : Six strains of primary HTM cells were isolated from cadaveric eyes with no history of eye disease and cultured under serum-starved condition in the presence or absence of 15% mechanical stretch, 1 cycle/second, for 24 hours using the computer controlled Flexcell Unit. Total RNA was isolated using Ambion mirVana miRNA Isolation kit. Expression of 800 human miRNAs was profiled using NanoString nCounter Human miRNA Assay kits. After quality check and normalization, differentially expressed miRNAs in the stretched condition were identified using Bioconductor Limma package. Droplet digital PCR was used to validate the differential expression of selected miRNAs.

Results : 373 miRNAs were detectably expressed in the cultured HTM cells. Of these miRNAs, 74 miRNAs (10 down- and 64 up-regulated) were differentially expressed in stretched cells (p≤0.05). The top 5 most significantly expressed miRNAs were miR-32-5p, miR-4286, miR-135-5p, miR-136-5p, miR-93-5p (p value= 1.2x10-5, 1.3x10-5, 4.5x10-5, 7x10-5, 2x10-4 respectively). The miRNAs with the greatest fold change in response to stretch included miR-4286, miR-32-5p, miR-136-5p, miR-93-5p, miR-135-5p, miR-21-5p, miR-19b-3p with log2 fold change ranging from 1.3 to 0.98. Consistent with previous reports, miR-100-5p, miR-27a-3p, miR-27b-3p, miR-24-3p, miR-16-5p, let-7f-5p and let-7i-5p were significantly upregulated in stretched versus control HTM cells. Ingenuity Pathway Analysis showed that these miRNAs are mostly associated with cellular functions related to proliferation, development, cell cycle regulation and cellular movement. Remarkably, the highly expressed miRNAs are predicted regulators of CDKN2A, a glaucoma-related gene shown to be upregulated in a rat glaucoma model.

Conclusions : Our miRNA profiling suggests a potential role of miRNAs in homeostatic responses of HTM cells to in vivo physiological mechanical stress. Further characterization of these miRNAs may advance our understanding of HTM-mediated regulation of aqueous outflow resistance and generate possible therapeutic targets to regulate intraocular pressure.

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

×
×

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.

×