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Modeling Keratoconus Using Induced Pluripotent Stem Cells.

Roy Joseph1, Om P Srivastava1, Roswell R Pfister2

  • 1Department of Optometry and Vision Science University of Alabama at Birmingham, Birmingham, Alabama, United States.

Investigative Ophthalmology & Visual Science
|July 13, 2016
PubMed
Summary
This summary is machine-generated.

Induced pluripotent stem cells (iPSC) from keratoconus (KC) patients were generated to model the disease. A down-regulated FGFR2-Pi3-Kinase pathway in KC iPSC suggests a mechanism for keratocyte apoptosis in KC.

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Area of Science:

  • Stem Cell Biology
  • Ophthalmology
  • Genetics

Background:

  • Keratoconus (KC) is a progressive thinning of the cornea.
  • Understanding the cellular mechanisms underlying KC is crucial for developing effective treatments.
  • Induced pluripotent stem cells (iPSC) offer a promising model for studying genetic eye diseases.

Purpose of the Study:

  • To establish a reliable in vitro model of keratoconus (KC) using patient-derived induced pluripotent stem cells (iPSC).
  • To investigate the molecular differences between KC and normal corneal cells at the stem cell level.
  • To identify potential pathways involved in the pathogenesis of KC.

Main Methods:

  • Human corneal fibroblasts from KC patients and healthy donors were reprogrammed into iPSC using a lentiviral vector.
  • iPSC were characterized for pluripotency markers (SSEA4, Oct4, Sox2, Tra-1-60, pax6) via immunofluorescence.
  • mRNA sequencing and Ingenuity Pathway Analysis (IPA) were performed to compare gene expression profiles between KC and normal iPSC.

Main Results:

  • Generated iPSC clones expressed key pluripotency and corneal markers.
  • Transcriptome analysis revealed significant gene expression differences, with 4300 genes showing a 2-fold change and 870 genes with q-value <0.05 in KC iPSC.
  • Fibroblast Growth Factor Receptor 2 (FGFR2) was significantly down-regulated in KC iPSC and KC-derived keratocytes, implicating the FGFR2-Pi3-Kinase pathway.

Conclusions:

  • A model for KC is proposed where inhibition of the FGFR2-Pi3-Kinase pathway impacts AKT phosphorylation, leading to reduced keratocyte survival signals.
  • This pathway inhibition may represent a key mechanism contributing to decreased cell survival and apoptosis observed in keratocytes from KC patients.
  • KC-derived iPSC and keratocytes exhibit impaired growth and proliferation, further supporting the identified pathway's role in the disease.