Duplication of TBK1 Stimulates Autophagy in iPSC-derived Retinal Cells from a Patient with Normal Tension Glaucoma

Budd A Tucker; Frances Solivan-Timpe; Ben R Roos; Kristin R Anfinson; Alan L Robin; Luke A Wiley; Robert F Mullins; John H Fingert

doi:10.4172/2157-7633.1000161

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Duplication of TBK1 Stimulates Autophagy in iPSC-derived Retinal Cells from a Patient with Normal Tension Glaucoma

Journal article

Open access

Duplication of TBK1 Stimulates Autophagy in iPSC-derived Retinal Cells from a Patient with Normal Tension Glaucoma

Budd A Tucker, Frances Solivan-Timpe, Ben R Roos, Kristin R Anfinson, Alan L Robin, Luke A Wiley, Robert F Mullins and John H Fingert

Journal of stem cell research & therapy, Vol.3(5), p.161

2014

DOI: 10.4172/2157-7633.1000161

PMCID: PMC4038935

PMID: 24883232

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Abstract

Duplication of the TBK1 gene causes normal tension glaucoma (NTG); however the mechanism by which this copy number variation leads to retinal ganglion cell death is poorly understood. The ability to use skin-derived induced pluripotent stem cells (iPSCs) to investigate the function or dysfunction of a mutant gene product in inaccessible tissues such as the retina now provides us with the ability to interrogate disease pathophysiology in vitro . iPSCs were generated from dermal fibroblasts obtained from a patient with TBK1 -associated NTG, via viral transduction of the transcription factors OCT4 , SOX2 , KLF4 , and c-MYC . Retinal progenitor cells and subsequent retinal ganglion cell-like neurons were derived using our previously developed stepwise differentiation protocol. Differentiation to retinal ganglion-like cells was demonstrated via rt-PCR targeted against TUJ1, MAP2, THY1, NF200, ATOH7 and BRN3B and immunohistochemistry targeted against NF200 and ATOH7. Western blot analysis demonstrated that both fibroblasts and retinal ganglion cell-like neurons derived from NTG patients with TBK1 gene duplication have increased levels of LC3-II protein (a key marker of autophagy). Duplication of TBK1 has been previously shown to increase expression of TBK1 and here we demonstrate that the same duplication leads to activation of LC3-II. This suggests that TBK1 -associated glaucoma may be caused by dysregulation (over-activation) of this catabolic pathway.

Stem Cells

Glaucoma

Retinal ganglion cells

TBK1

Autophagy

iPSC

Details

Title: Subtitle: Duplication of TBK1 Stimulates Autophagy in iPSC-derived Retinal Cells from a Patient with Normal Tension Glaucoma
Creators: Budd A Tucker - Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA
Frances Solivan-Timpe - Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA
Ben R Roos - Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA
Kristin R Anfinson - Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA
Alan L Robin - Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA
Luke A Wiley - Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA
Robert F Mullins - Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA
John H Fingert - Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA
Resource Type: Journal article
Publication Details: Journal of stem cell research & therapy, Vol.3(5), p.161
DOI: 10.4172/2157-7633.1000161
PMID: 24883232
PMCID: PMC4038935
ISSN: 2157-7633
eISSN: 2157-7633
Language: English
Date published: 2014
Academic Unit: Ophthalmology and Visual Sciences
Record Identifier: 9983979962702771

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