Elsevier

Stem Cell Research

Volume 21, May 2017, Pages 58-70
Stem Cell Research

Patient-specific induced pluripotent stem cells to evaluate the pathophysiology of TRNT1-associated Retinitis pigmentosa

https://doi.org/10.1016/j.scr.2017.03.005Get rights and content
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open access

Highlights

  • Utilizing patient-derived iPSCs to study RP.

  • Generating iPSC-derived retinal organoids from patients with molecularly confirmed TRNT1-associated RP.

  • Mutations in TRNT1 disrupt autophagy in patient-specific retinal cells.

Abstract

Retinitis pigmentosa (RP) is a heterogeneous group of monogenic disorders characterized by progressive death of the light-sensing photoreceptor cells of the outer neural retina. We recently identified novel hypomorphic mutations in the tRNA Nucleotidyl Transferase, CCA-Adding 1 (TRNT1) gene that cause early-onset RP. To model this disease in vitro, we generated patient-specific iPSCs and iPSC-derived retinal organoids from dermal fibroblasts of patients with molecularly confirmed TRNT1-associated RP. Pluripotency was confirmed using rt-PCR, immunocytochemistry, and a TaqMan Scorecard Assay. Mutations in TRNT1 caused reduced levels of full-length TRNT1 protein and expression of a truncated smaller protein in both patient-specific iPSCs and iPSC-derived retinal organoids. Patient-specific iPSCs and iPSC-derived retinal organoids exhibited a deficit in autophagy, as evidenced by aberrant accumulation of LC3-II and elevated levels of oxidative stress. Autologous stem cell-based disease modeling will provide a platform for testing multiple avenues of treatment in patients suffering from TRNT1-associated RP.

Keywords

Induced pluripotent stem cells
Retinitis pigmentosa
TRNT 1
Retinal organoids
Autophagy
Oxidative stress

Abbreviations

RP
Retinitis pigmentosa
TRNT1
tRNA Nucleotidyl Transferase, CCA-Adding 1
iPSC
induced pluripotent stem cells
CSA
congenital sideroblastic anemias
SIFD
B-cell immunodeficiency, periodic fevers and development delay
UPR
unfolded protein response
ER
endoplasmic reticulum
BIP
binding immunoglobulin protein
KLF4
kruppel-like factor 4
OCT4
octamer-binding transcription factor 4
SOX2
sex determining region Y-box 2
c-MYC
V-myc avian myelocytomatosis viral oncogene homolog
NANOG
nanog homeobox
LC3
microtubule-associated protein 1A/1B-light chain 3
LAMP1
lysosomal-associated membrane protein 1
GAPDH
glyceraldehyde 3-phosphate dehydrogenase

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