June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Entry into the pluripotency continuum impacts retinal cell fate decisions of human iPSCs.
Author Affiliations & Notes
  • Jessica A Cooke
    Ophthalmology, University of Iowa Hospitals and Clinics, Iowa City, Iowa, United States
  • Andrew P Voigt
    Ophthalmology, University of Iowa Hospitals and Clinics, Iowa City, Iowa, United States
  • Michael A Collingwood
    Enzyme Development, Integrated DNA Technologies Inc, Coralville, Iowa, United States
  • Nicholas Edwin Stone
    Ophthalmology, University of Iowa Hospitals and Clinics, Iowa City, Iowa, United States
  • Sarah Carrothers
    Ophthalmology, University of Iowa Hospitals and Clinics, Iowa City, Iowa, United States
  • S Scott Whitmore
    Ophthalmology, University of Iowa Hospitals and Clinics, Iowa City, Iowa, United States
  • Adam P DeLuca
    Ophthalmology, University of Iowa Hospitals and Clinics, Iowa City, Iowa, United States
  • Edwin M Stone
    Ophthalmology, University of Iowa Hospitals and Clinics, Iowa City, Iowa, United States
  • Robert Mullins
    Ophthalmology, University of Iowa Hospitals and Clinics, Iowa City, Iowa, United States
  • Budd A. Tucker
    Ophthalmology, University of Iowa Hospitals and Clinics, Iowa City, Iowa, United States
  • Footnotes
    Commercial Relationships   Jessica Cooke None; Andrew Voigt None; Michael Collingwood None; Nicholas Stone None; Sarah Carrothers None; S Scott Whitmore None; Adam DeLuca None; Edwin Stone None; Robert Mullins None; Budd Tucker None
  • Footnotes
    Support  Institute for Vision Research
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 3174. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Jessica A Cooke, Andrew P Voigt, Michael A Collingwood, Nicholas Edwin Stone, Sarah Carrothers, S Scott Whitmore, Adam P DeLuca, Edwin M Stone, Robert Mullins, Budd A. Tucker; Entry into the pluripotency continuum impacts retinal cell fate decisions of human iPSCs.. Invest. Ophthalmol. Vis. Sci. 2023;64(8):3174.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : To determine how an hiPSC’s transcriptomic profile developed during reprograming can be altered to maximize retinal cell differentiation capacity.

Methods : Human dermal fibroblasts that were reprogrammed via Sendai virus transduction were subjected to RNA sequencing at 0- and 7-days post-differentiation. Retinal structures were quantified for retinal phenotype microscopically. Analysis was focused on identifying genes differentially expressed between good retinal producers (40-80% of organoids showing a retinal phenotype, n=8) and poor retinal producers (<30% of organoids showing retinal phenotype, n =7). Differential gene expression was validated in a masked fashion.

Results : By 7 days of differentiation, when embryoid bodies are indistinguishable, significant differences in gene expression could be detected between good vs poor retinal producers. Ingenuity pathway analysis revealed perturbations in both OCT4 and SOX2 effector gene expression, with expression of many of the factors being maintained in poor producers indicating delayed suppression of pluripotency. Of the differentially expressed genes evaluated, seven (NODAL, CER1, EN1, GBX2, MIXL1, GSC, and EOMES) were found to be down regulated in cell lines that produced greater than 10% retinal organoids. Three genes (LEFTY2, ANXA1, and FOXD3) were found to be expressed at high levels in cell lines that produced less than 20% retinal organoids, and four genes (RAX, LHX2, VSX2, SIX6) known to be markers of retinal differentiation, were expressed at low levels in the poorest producers with a gradual increase in expression as the percent of retinal organoids per culture increased.

Conclusions : A signature gene expression profile was used to develop a qPCR panel with the ability to predict retinal differentiation propensity at the earliest stages of differentiation. Modulation of these key factors may assist in enhancement of retinal differentiation capacity.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

×
×

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.

×