September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
PAX6 Isoforms, along with Reprogramming Factors, Differentially Regulate the Induction of Cornea-specific Genes
Author Affiliations & Notes
  • Yuzuru Sasamoto
    Ophthalmology, Osaka University Medical School, Suita, Japan
  • Ryuhei Hayashi
    Stem Cell and Applied Medicine, Osaka University Medical School, Suita, Japan
  • Sung-Joon Park
    The Institute of Medical Science, The University of Tokyo, Minato-ku, Japan
  • Mihoko Saito-Adachi
    The Institute of Medical Science, The University of Tokyo, Minato-ku, Japan
    Division of Cancer Genomics, National Cancer Center Research Institute, Chuo-ku, Japan
  • Yutaka Suzuki
    Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa, Japan
  • Satoshi Kawasaki
    Ophthalmology, Osaka University Medical School, Suita, Japan
  • Andrew J Quantock
    School of Optometry and Vision Sciences, Cardiff University, Cardiff, United Kingdom
  • Kenta Nakai
    The Institute of Medical Science, The University of Tokyo, Minato-ku, Japan
  • Motokazu Tsujikawa
    Ophthalmology, Osaka University Medical School, Suita, Japan
  • Kohji Nishida
    Ophthalmology, Osaka University Medical School, Suita, Japan
  • Footnotes
    Commercial Relationships   Yuzuru Sasamoto, None; Ryuhei Hayashi, None; Sung-Joon Park, None; Mihoko Saito-Adachi, None; Yutaka Suzuki, None; Satoshi Kawasaki, None; Andrew Quantock, None; Kenta Nakai, None; Motokazu Tsujikawa, None; Kohji Nishida, None
  • Footnotes
    Support  Japan Eye Bank Association
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 4906. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to Subscribers Only
      Sign In or Create an Account ×
    • Get Citation

      Yuzuru Sasamoto, Ryuhei Hayashi, Sung-Joon Park, Mihoko Saito-Adachi, Yutaka Suzuki, Satoshi Kawasaki, Andrew J Quantock, Kenta Nakai, Motokazu Tsujikawa, Kohji Nishida; PAX6 Isoforms, along with Reprogramming Factors, Differentially Regulate the Induction of Cornea-specific Genes. Invest. Ophthalmol. Vis. Sci. 2016;57(12):4906.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : We developed the transplantation of cultured autologous oral mucosal epithelial sheets for eyes with bilateral limbal stem cell deficiencies. This approach has shown some promise; however, the reconstructed ocular surface does not allow a full, long-term improvement of visual acuity, and is prone to problems associated with neovascularization. Presumably, this is due to the incomplete corneal phenotype of the reconstructed epithelium, owing at least to the fact that the oral mucosal epithelium lacks corneal epithelial specific genes, such as PAX6 and KRT12. To reduce such limitations in our ability to reconstruct and maintain the ocular surface, we tested the function of two PAX6 isoforms in the human oral mucosal epithelial cells.

Methods : Two PAX6 isoforms (PAX6a and PAX6b), along with Yamanaka four factors (OCT4, SOX2, KLF4, c-MYC), were transduced into human oral mucosal epithelial cells by lentiviruses. Three days after transduction, we measured the expression of corneal epithelium specific genes, keratin 3 (KRT3) and keratin 12 (KRT12). Reporter assay and single-cell gene expression analysis were also performed.

Results : The highest induction of KRT12 was obtained by the combinational transduction of PAX6b, OCT4 and KLF4. KRT3 was induced by PAX6a solely and it was enhanced when combined with KLF4 transduction. Reporter assay showed that PAX6a targeted the upstream region of KRT3 gene. Single-cell gene expression analysis revealed that KRT12 induction required high expression of transgenes. KRT3 and KRT12 were preferentially induced in surface ectoderm-derived cells when compared to the other cell source.

Conclusions : Two PAX6 isoforms differentially and cooperatively regulate the expression of genes specific to the structure and function of the corneal epithelium, particularly KRT3 and KRT12.

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

×
×

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.

×