September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Gene Expression Levels in Microsurgical Specimens from Patients with Glaucoma
Author Affiliations & Notes
  • Dennis Mock
    Stein Eye Institute , LA, California, United States
    David Geffen School of Medicine at UCLA, Los Angeles, California, United States
  • Rajendra K Gangalum
    Stein Eye Institute , LA, California, United States
  • Suraj P Bhat
    Stein Eye Institute , LA, California, United States
    Molecular Biology Institute & Brain Research Institute, Los Angeles, California, United States
  • Anne L Coleman
    Stein Eye Institute , LA, California, United States
    Epidemiology, Jonathan & Karin Fieldings School of Public Heath, Los Angeles, California, United States
  • Footnotes
    Commercial Relationships   Dennis Mock, None; Rajendra Gangalum, None; Suraj Bhat, None; Anne Coleman, None
  • Footnotes
    Support  This work was supported by an Oppenheimer Foundation Grant to D.M. and A.L.C.
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 6026. doi:
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    • Get Citation

      Dennis Mock, Rajendra K Gangalum, Suraj P Bhat, Anne L Coleman; Gene Expression Levels in Microsurgical Specimens from Patients with Glaucoma. Invest. Ophthalmol. Vis. Sci. 2016;57(12):6026.

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

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Abstract

Purpose : We investigate a genomic basis of the gene expression in the trabecular meshwork/extracellular matrix of the eye and its potential association with primary open-angle glaucoma , a blinding eye disease involving the degeneration of retinal ganglion cells and the optic nerve.

Methods : Microsurgical tissue samples (~ 0.1 mm block or less of extracellular/trabecular meshwork (TM-ECM)) were collected from 28 patients undergoing a trabeculectomy to reduce intraocular pressure (IOP) for primary open-angle glaucoma (POAG). Using a multiplex high-throughput microfluidic real-time RT-qPCR system, we examined the individual RNA isolates from these samples for the expression of 48 genes known to be associated with the trabecular meshwork/extracellular matrix and elevated IOP. For each gene, we determined its transcriptional range of expression (threshold cycles, Ct ) and the frequency distribution for the gene detection across all the samples (expression heterogeneity).

Results : Profiles of RNA transcripts exhibited a high efficiency/sensitivity (mean efficiency > 95.0%, ΔCt ~9-35) and specificity (coefficient of variation<0.05, amplicon peak location for melting curve). The data showed a very high level of RNA transcript heterogeneity for various genes across the individual samples (32.1 + 12.7 %). Genes such as optineurin (OPTN) and myocilin (MYOC), previously associated with glaucoma from genome-wide association (GWA) and genetic linkage studies, were among those transcripts having the highest range of expression (differ by ΔCt> 25) and expression heterogeneity (~50% ).

Conclusions : This study reveals the presence of potentially useful biological/molecular information in the discarded microsurgical tissue removed during a trabeculectomy for the treatment of glaucoma. Transcription readouts from primary tissue may provide a genomic "snaphot" for the dynamic progression of the disease.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

 

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